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1.
Rev. Hosp. Ital. B. Aires (2004) ; 42(3): 143-151, sept. 2022. graf, ilus, tab
Article in Spanish | LILACS, BINACIS, UNISALUD | ID: biblio-1396799

ABSTRACT

Introducción: en diciembre del año 2019 surgió en China una neumonía viral; el virus fue identificado como un coronavirus SARS-CoV-2, que se propagó rápidamente de tal manera que se convirtió en pandemia. La alta contagiosidad y la presencia de portadores asintomáticos dificultaron el diagnóstico de la infección y la toma de decisiones sanitarias. Objetivo: el objetivo de esta revisión bibliográfica es presentar y describir las principales técnicas utilizadas actualmente para el diagnóstico de COVID-19 y establecer su relación con los conocimientos de distintas disciplinas y tecnologías emergentes que confluyen en la Biotecnología bioquímico-farmacéutica orientada a la Salud humana. Metodología: se realizó una revisión de la bibliografía disponible en PubMed a partir de enero de 2020 sobre las pruebas diagnósticas que se encuentran actualmente en uso, en el ámbito sanitario, para la detección y seguimiento de la enfermedad COVID-19. También se realizaron búsquedas a través de Google y Google Académico para publicaciones de organismos de Salud en referencia a métodos diagnósticos. Resultados: se presenta una importante cantidad de pruebas diagnósticas, basadas en diferentes tecnologías, que desempeñan un papel clave en la pandemia de COVID-19. Algunas de ellas muy sofisticadas, como la secuenciación genómica de próxima generación, otras más estándar, pero igualmente robustas, como la reacción en cadena de la polimerasa (PCR). También otras adaptadas para el brote pandémico, como la amplificación isotérmica de ácidos nucleicos mediada por bucle. Todas las mencionadas se consideran de tipo molecular, pero también existen las pruebas serológicas, como ELISA, que incluyen ensayos en plasma o de tipo inmunológico. Estas sirven para detectar anticuerpos frente a la exposición al virus o antígenos en personas potencialmente infectadas. Conclusiones: los procesos de investigación y desarrollo biotecnológicos aplicados al diagnóstico y los conocimientos científicos previos permitieron una respuesta tanto nacional como internacional rápida y eficaz en medio de una inédita pandemia global. En esta revisión destacamos las principales técnicas, en qué estadio se deben usar y qué información nos aportan. (AU)


Introduction: in December 2019, a viral pneumonia emerged in China, identifying the virus as a SARS-CoV-2 coronavirus, which spread rapidly in such a way that it became a pandemic. The high contagiousness and the presence of asymptomatic carriers make difficulted to diagnose the infection and to make health decisions. Target: the objective of this review is to present and describe the main techniques currently used for the diagnosis of COVID-19, and to establish their relationship with the knowledge of different disciplines and emerging technologies that converge in biochemical-pharmaceutical biotechnology oriented to human health. Methodology: a review of the literature available in Pubmed from January 2020 on the diagnostic tests that are currently in use in the health field, for the detection and monitoring of COVID-19 disease, was carried out. Searches were also carried out through Google and Google Scholar for publications of Health organizations in reference to diagnostic methods. Results: a significant number of diagnostic tests are presented, based on different technologies, which play a key role in the COVID-19 pandemic. Some of them are very sophisticated, such as next-generation genomic sequencing, others more standard, but equally robust, such as polymerase chain reaction. Also others adapted for the pandemic outbreak such as loop-mediated isothermal amplification of nucleic acids. All of the aforementioned are considered molecular, but there are also serological tests, such as ELISA, which include plasma or immunological tests. These serve to detect antibodies against exposure to the virus or antigens in potentially infected people. Conclusions: biotechnological research and development processes applied to diagnosis and previous scientific knowledge allowed a rapid and effective national and international response in the midst of an unprecedented global pandemic. In this review we highlight the main techniques, at what stage they should be used and what information they provide us. (AU)


Subject(s)
Humans , Biotechnology/trends , COVID-19 Testing/methods , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , Enzyme-Linked Immunosorbent Assay , Biomarkers , Sensitivity and Specificity , Diagnostic Techniques and Procedures , COVID-19 Nucleic Acid Testing , COVID-19 Serological Testing
2.
Rev. med. vet. zoot ; 69(2): 155-165, mayo-ago. 2022. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1395018

ABSTRACT

Resumen Los virus de inmunodeficiencia y leucemia felina representan un problema de gran envergadura para los felinos domésticos debido a la multiplicidad de sintomatologías que manifiestan. El objetivo del presente estudio fue establecer, retrospectivamente, la prevalencia en la presentación de ViLeF y VIF en pacientes de seis clínicas de pequeños animales en Bogotá y Chía, en relación con factores como su edad, raza y género. Se realizó un estudio transversal y retrospectivo, mediante la recopilación de datos de 1.014 historias clínicas de pacientes felinos que ingresaron a seis clínicas de la ciudad de Bogotá y Chía, para determinar la prevalencia de VIF y ViLeF y la asociación de estas con factores como edad, género y raza, entre 2015 y 2019, a través de la prueba OR. La detección de los virus se realizó mediante una prueba rápida basada en inmunocromatografía. La mayor prevalencia para cada enfermedad por año fue: 12,3% para VIF en 2012 y 18% para ViLeF en 2019. Los machos presentaron mayores seroprevalencias para ambas enfermedades durante la mayoría los años evaluados. Factores como raza (criolla: VIF: 1,85; ViLeF: 2,01), género (macho: VIF: 1,53 OR; ViLeF: 1,64) y edad (> 7 años: VIF: 3,82; ViLeF: 3,21) se relacionaron positivamente con la presentación de ambas enfermedades en la población felina evaluada.


Abstract Immunodeficiency virus and feline leukemia virus represent major problems for domestic felines due to the multiplicity of symptoms they manifest. The objective of the present study was to establish, retrospectively, the prevalence in the presentation of FeLV and FIV in patients from six small animal clinics in Bogota and Chia, related to factors such as age, race, and gender. A cross-sectional and retrospective study was carried out, collecting data from 1.014 clinical records of feline patients who were admitted to six clinics in the city of Bogota and Chia, to determine the prevalence of FIV and FeLV and their association with factors such as age, gender, and race, between 2015 and 2019 through the OR test. The detection of the viruses was carried out through a rapid test based on immunochromatography. The highest prevalence for each disease per year was 12,3% for FIV in 2012 and 18% for FeLV in 2019. Males presented higher seroprevalences for both diseases during most of the years evaluated. Factors such as race (Creole: FIV: 1,85; FeLV: 2,01), gender (male: FIV: 1.53 OR, FeLV: 1,64), and age (> 7 years: FIV: 3.82; FeLV: 3.21) were positively related to the presentation of both diseases in the feline population evaluated.


Subject(s)
Animals , Cats , Viruses , Enzyme-Linked Immunosorbent Assay , Leukemia , Chronic Disease , Disease , Chromatography, Affinity , Immunodeficiency Virus, Feline , Leukemia Virus, Feline , Diagnosis , Retroviridae , Hospitals, Animal
3.
Rev. bras. ciênc. vet ; 29(3): 130-134, jul./set. 2022. il.
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1411231

ABSTRACT

A brucelose é uma doença bacteriana de grande importância para a economia pecuária e para a saúde pública por se tratar de uma zoonose. É uma doença infecto-contagiosa que tem com agente etiológico bactérias do gênero Brucella. Em bovinos, as espécies do gênero é a Brucella abortus, que são cocobacilos gram negativo, intracelulares facultativos, imóveis e não esporulado. A infecção apresenta evolução crônica e acomete animais de todas as idades, sendo mais frequente em indivíduos sexualmente maduros. O objetivo desse trabalho é investigar, por meio da sorologia para brucelose bovina, utilizando a técnica do ELISA indireto, amostras de animais reagentes abatidos em frigoríficos inspecionados no estado da Bahia. Foram utilizados 666 animais, selecionados aleatoriamente no momento do abate. O sangue foi coletado com finalidade de obtenção de soro, todas as amostras foram submetidas à prova de triagem do Antígeno Acidificado Tamponado (AAT), prova do 2mercaptoetanol (2-ME) e ELISA Indireto. Das amostras reagentes no teste do AAT, obteve-se uma prevalência estimada em 1,2%. A prevalência no teste do ELISA foi de 13,21% (n=86). Esse resultado sugere a ocorrência de falsos negativos quando se utiliza a prova do antígeno acidificado tamponado.


Brucellosis is a bacterial disease of great importance to the livestock economy and to public health because it is a zoonosis. It is an infectious disease that has etiologic agent with bacteria of the genus Brucella. In cattle, the species of the genus Brucella is Brucella abortus that are gram negative, facultative intracellular, real estate and not sporulated. The infection presents chronic and affects animals of all ages, being more frequent in sexually mature individuals. This study aimed to investigate through serology for brucellosis, using the technique of indirect ELISA, samples from positive animals slaughtered in slaughterhouses inspected in the state of Bahia. A total of 666 animals were used, randomly selected at the time of slaughter. Blood was collected in order to obtain serum, all samples were subjected to a screening test Antigen Buffered Acidified (AAT), proof of 2-mercaptoethanol (2-ME) and Indirect ELISA. Of reagents in the test samples of AAT obtained an estimated prevalence of 1.2%. The prevalence in the ELISA test was 13.21% (n = 86). This result suggests the occurrence of false negatives when using the buffered acidified antigen test.


Subject(s)
Animals , Cattle , Brucella abortus , Brucellosis, Bovine/diagnosis , Cattle/abnormalities , Enzyme-Linked Immunosorbent Assay/veterinary , Bacterial Zoonoses/diagnosis , Prevalence
4.
Rev. cuba. med ; 61(2): e2637, abr.-jun. 2022. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1408993

ABSTRACT

Introducción: Los autoanticuerpos anti-insulina (AAI) representan un marcador serológico de la diabetes tipo 1 (DT1). El significado clínico de los AAI aún no ha sido determinado en la población cubana. Objetivo: Determinar el valor clínico de AAI en pacientes con DT1. Métodos: Se determinaron los niveles séricos de AAI por el ensayo inmuno-adsorbente ligado a enzima (ELISA) en 33 pacientes adultos con DT1, 78 pacientes con otras condiciones endocrinas (CEE) como diabetes tipo 2, tiroiditis de Hashimoto e hiperinsulinemia, y 49 controles normales (CN). El valor de corte se determinó con el análisis de las curvas características operativas del receptor (COR) (ROC por sus siglas en inglés). Se utilizaron pruebas no paramétricas para comparar los niveles de AAI de pacientes con DT1, CEE y CN, y determinar la correlación entre AAI y la edad. Resultados: El valor de corte óptimo de AAI para DT1 fue el índice de 1,05, con sensibilidad de 45,5 por ciento, especificidad de 81,6 por ciento, razón de verosimilitud positiva de 2,47, y razón de verosimilitud negativa de 0,67. Los niveles de AAI en DT1 (índice de 0,97) fueron significativo, más altos que los de CN (índice de 0,70; p=0,020) y los de CEE (índice de 0,63; p= 0,009). Los niveles de AAI resultaron inversamente proporcionales a la edad en pacientes diabéticos ( =-0,252; p=0,030). Conclusiones: Los pacientes con DT1 se distinguieron por niveles más altos de AAI, aunque la presencia de estos anticuerpos no fue exclusiva de DT1. Los niveles de AAI dependieron de la edad en los pacientes diabéticos(AU)


Introduction: Anti-insulin autoantibodies (AAI) represent a serological marker of type 1 diabetes (T1D). The clinical significance of AAIs has not yet been determined in the Cuban population. Objective: To determine the clinical value of AAI in patients with T1D. Methods: AAI serum levels were determined by enzyme-linked immunosorbent assay (ELISA) in 33 adult patients with T1D, 78 patients with other endocrine conditions (CEE) such as type 2 diabetes, Hashimoto's thyroiditis, and hyperinsulinemia, and 49 normal controls (CN). The cut-off value was determined by receiver operating characteristic (ROC) curve analysis. Nonparametric tests were used to compare the AAI levels of patients with T1D, CEE, and CN, and to determine the correlation between AAI and age. Results: AAI optimal cut-off value for T1D was the index of 1.05, with 45.5 percent of sensitivity, 81.6 percent specificity, 2.47 positive likelihood ratio, and 0.67 negative likelihood ratio. AAI levels in DT1 (index of 0.97) were significant, higher than those of CN (index of 0.70; p= 0.020) and CEE levels (index of 0.63; p= 0.009). AAI levels were inversely proportional to age in diabetic patients (ρ = -0.252; p=0.030). Conclusions: Patients with T1D were distinguished by AAI higher levels, although the presence of these antibodies was not exclusive to T1D. AAI levels depended on age in diabetic patients(AU)


Subject(s)
Humans , Male , Female , Autoantibodies , Enzyme-Linked Immunosorbent Assay/methods , Diabetes Mellitus, Type 1/epidemiology , Cuba , Insulin Antibodies
5.
Rev. med. vet. zoot ; 69(1): 11-18, ene.-abr. 2022. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1389163

ABSTRACT

RESUMEN La leucemia viral felina (ViLeF) es una enfermedad retroviral letal, de una elevada prevalência en Colombia, que afecta a felinos de diferentes edades y sexos. El objetivo de esta investigación fue determinar la frecuencia por serodiagnóstico de ViLeF en felinos del centro integral de bienestar animal Ceiba, ubicado en Rionegro, Antioquia (Colombia), en 2020. Para ello, se realizó un estudio descriptivo longitudinal de serofrecuencia de ViLeF desde enero hasta diciembre de 2020. Fueron muestreados 92 gatos, a los cuales se les efectuó una prueba p27 por inmunoensayo comercial Elisa (Idexx©, Snap Combo Plus®, Maine, EE. UU.). La frecuencia de felinos positivos fue 30/92 (32,60%) y el mes de mayo fue el de mayor frecuencia (9,78%). Los machos positivos fueron 17/92 (18,47%) y las hembras 13/92 (14,13%). La edad promedio de seropositividad fue 2,14 años. La frecuencia de ViLeF en 2020 para Ceiba, Rionegro (Colombia) es de 32,60%, un valor elevado con respecto a descripciones en otros albergues para felinos. ViLeF es una enfermedad que está siendo reportada con mayor frecuencia en Colombia, debido a que las medidas de prevención no se están adoptando rutinariamente.


ABSTRACT Feline viral leukemia (ViLeF) is a lethal retroviral disease with a high prevalence in Colombia that affects felines of different ages and sexes. The purpose of this investigation was to determine the frequency by serodiagnosis of ViLeF in felines of the integral center of animal welfare Ceiba, located in Rionegro, Antioquia (Colombia), during 2020. For that, a longitudinal descriptive study of ViLeF serofrequency from were made January to December 2020. 92 cats were sampled, which were tested for p27 by commercial Elisa immunoassay (Idexx©, Snap Combo Plus®, Maine, USA). The frequency of positive felines was 30/92 (32,60%). May was the month with the highest frequency (9,78%). The positivity frequency for males was 17/92 (18,47%) and the frequency for females 13/92 (14,13%). The main age of seropositivity was 2,14 years. The frequency of ViLeF in 2020 for Ceiba, Rionegro (Colombia) is 32,60%. This is a high value in comparison to descriptions in other shelters for felines. ViLeF, in Colombia, is a disease that has been reported with more frequency because prevention measures are not being adopted routinely.


Subject(s)
Animals , Cats , Enzyme-Linked Immunosorbent Assay , Morbidity , Chromatography, Affinity , Leukemia, Feline , Leukemia Virus, Feline , Felidae , Immunoassay , Serologic Tests , Disease , Prevalence
6.
Article in English | LILACS-Express | LILACS, CUMED | ID: biblio-1410302

ABSTRACT

In Egypt, the lyophilized live attenuated sheep pox virus vaccine has been used for the vaccination of cattle against lumpy skin disease virus to control its economic impact on livestock industry. In this endeavor, we validate the efficacy of Carbopol® as a stabilizer and adjuvant to enhance immunogenicity of such a heterologous sheep pox virus vaccine against lumpy skin disease. Lyophilization of sheep pox virus vaccine stabilized with Carbopol® produced better physical and antigenic properties than freeze-drying with lactalbumin/sucrose stabilizer; this was manifested by superior disc uniformity, thermo-stability at 37oC, and less reduction in virus titer. Immunization of calves' groups with variable sheep pox vaccine doses containing different Carbopol® concentrations revealed that 103.5 TCID50 of sheep pox virus vaccine enclosing 0.5 percent Carbopol® is the field dose of choice. Moreover, it induced protective serum neutralizing index of 2.5 and a ELISA S/P ratio of 36, by the 4th week post vaccination. Besides, the inclusion of 0.5 percent Carbopol® in formulation of the sheep pox virus vaccine was safe in bovines and enhanced cellular immune response to lumpy skin disease virus, as evidenced by increased T cell proliferation. Hence, it is recommended to use Carbopol® as 0.5 percent in preparation of live attenuated sheep pox virus vaccine to confer better protection against lumpy skin disease virus infection(AU)


En Egipto, la vacuna atenuada liofilizada contra el virus de la viruela ovina ha sido utilizado para la vacunación del ganado, contra el virus de la dermatosis nodular contagiosa, para controlar su impacto económico en la industria ganadera. En este trabajo, validamos la eficacia del Carbopol®, como estabilizador y adyuvante, para mejorar la inmunogenicidad de dicha vacuna heteróloga contra la dermatosis nodular contagiosa. La liofilización de la vacuna contra el virus de la viruela ovina estabilizada con Carbopol®, resultó en mejores propiedades físicas y antigénicas que la liofilización con el estabilizador de lactoalbúmina/sacarosa; lo anterior se manifestó en la uniformidad superior del disco, la termoestabilidad a 37°C y la menor reducción del título del virus. La inmunización de grupos de terneros con dosis variables de vacuna contra el virus de la viruela ovina, que contenían diferentes concentraciones de Carbopol®, reveló que la dosis de campo de elección fue 103,5 TCID50 de la vacuna contra el virus de la viruela ovina conteniendo 0,5 por ciento de Carbopol®, la que indujo un índice de neutralización sérica protectora de 2,5 y una relación S/P de ELISA de 36 a la cuarta semana después de la vacunación. Además, la inclusión de Carbopol® al 0,5 por ciento en la formulación de la vacuna contra el virus de la viruela ovina fue segura en los bovinos y potenció la respuesta inmunitaria celular contra el virus de la dermatosis nodular contagiosa, como lo demuestra el aumento de la proliferación de células T. Por lo tanto, se recomienda el uso de Carbopol® al 0,5 por ciento en la preparación de la vacuna viva atenuada contra el virus de la viruela ovina para conferir una mejor protección contra la infección por el virus de la dermatosis nodular contagiosa(AU)


Subject(s)
Animals , Enzyme-Linked Immunosorbent Assay/methods , Capripoxvirus/pathogenicity , Reference Drugs , Lumpy skin disease virus/pathogenicity , Vaccines , Vaccines, Attenuated/therapeutic use , Egypt
7.
Rev. chil. infectol ; 39(1): .45-52, feb. 2022. ilus, tab
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1388331

ABSTRACT

INTRODUCCIÓN: La enfermedad de Chagas es una infección parasitaria crónica sistémica, de importancia global, causada por Trypanosoma cruzi. OBJETIVO: Determinar la prevalencia de anticuerpos contra T cruzi en mujeres embarazadas en el estado de Morelos, México. MATERIALES Y MÉTODOS: Se analizaron 1.620 sueros de mujeres embarazadas mediante dos pruebas serológicas: ELISAc (antígeno crudo nativo) y ELISAr (antígeno recombinante, no nativo). Las muestras reactivas se analizaron posteriormente mediante hemaglutinación indirecta (HAI). Se utilizaron dos enfoques de detección, en paralelo (son positivas las muestras reactivas por cualquier método) y en serie (son positivas las muestras confirmadas por HAI). Se evaluaron factores sociodemográficos y de salud asociados a la presencia de anticuerpos contra T. cruzi mediante razones de momios al 95%. RESULTADOS: Se obtuvo una seroprevalencia de 4,87% con el diagnóstico en paralelo y de 0,43% en serie. A partir de los resultados en paralelo las mujeres que fueron atendidas en los hospitales generales de Tetecala y Jojutla tuvieron, respectivamente, 2,2 y 2,0 veces mayor posibilidad de presentar anticuerpos contra T cruzi con respecto a las mujeres que fueron atendidas en el Hospital General de Cuautla. CONCLUSIÓN: La prevalencia de anticuerpos contra T cruzi en mujeres embarazadas en el estado de Morelos fluctuó entre 0,43 y 4,87%, según el antígeno y el abordaje utilizado. Es necesario continuar con la vigilancia de la seroprevalencia de anticuerpos contra T cruzi en mujeres embarazadas en el estado de Morelos, México, con las técnicas de mayor sensibilidad y especificidad disponibles.


BACKGROUND: Chagas disease is a globally important chronic systemic parasitic infection caused by Trypanosoma cruzi. AIM: To determine the prevalence of antibodies against T cruzi in pregnant women from the state of Morelos, México. METHODS: 1,620 sera from pregnant women were analyzed using two serological tests: ELISAc (native crude antigen) and ELISAr (recombinant, non-native antigen). Reactive samples were subsequently analyzed by indirect hemagglutination (IHA). Two detection approaches were used, in parallel (reactive samples by any method are positive) and serial (samples confirmed by IHA are positive). Sociodemographic and health factors associated with the presence of antibodies against T cruzi were evaluated using 95% odds ratios. RESULTS: A seroprevalence of 4.87% was obtained with parallel diagnosis and 0.43% in series. From the parallel results, the women who were attended at the general hospitals of Tetecala and Jojutla had respectively 2.2 and 2.0 times greater chance of presenting antibodies against T cruzi compared to the women who were attended at the General Hospital of Cuautla. CONCLUSION: The prevalence of antibodies against T cruzi in pregnant women from the state of Morelos fluctuated between 0.43 and 4.87%, depending on the antigen and the approach used. It is necessary to continue with the surveillance of the seroprevalence of antibodies against T cruzi in pregnant women from the state of Morelos, Mexico, using the techniques with the highest sensitivity and specificity available.


Subject(s)
Humans , Female , Pregnancy , Trypanosoma cruzi , Chagas Disease/diagnosis , Chagas Disease/epidemiology , Enzyme-Linked Immunosorbent Assay/methods , Antibodies, Protozoan , Seroepidemiologic Studies , Pregnant Women , Mexico/epidemiology
8.
Rev. bras. ciênc. vet ; 29(1): 41-45, jan./mar. 2022. il.
Article in English | LILACS, VETINDEX | ID: biblio-1393235

ABSTRACT

This study aimed to promote the standardization of an indirect, enzyme-linked immunosorbent assay (ELISA) for the serological detection of B. anserina in Gallus gallusdomesticus. An aliquoted sera from vaccinated chicken with B. anserina antigen (GI), experimental infected chickens with B. anserina (GII) and rustic poultry rearing of G. gallus (GIII) were tested with in-house ELISA developed to detect serum antibodies against B. anserina in G. gallus domesticus. On average, the experimentally infected chickens became positive at 9 DPI a mean ± standard deviation (SD) ODI value of 163.11 ± 70.65. The highest observed Optical Density Index (ODI) was 372.54 ± 132.39, at 26 DPI, and the highest overall ODI value was 626.51. The vaccinated chickens became positive between 8 and 10 DPV, with an ODI of 245.59 at 10 DPV, with an overall maximum ODI of 543.13. A total of 108 blood samples were collected from poultry raised on rustic farms. Of the total samples collected, 58.33% (63/108) were considered positive for B. anserina. The maximum ODI found among these rustic chickens was 283.24. This stardardization provided a sensitivity and specificity of 100%.


Este estudo teve como objetivo promover a padronização de um ensaio imunoenzimático indireto (ELISA) para a detecção sorológica de Borrelia anserina em Gallus gallus domesticus. Um frango vacinado com antígeno de B. anserina (GI), frangos infectados experimentalmente com B. anserina (GII) e frangos criados de forma rústica (GIII) foram testados com ELISA indireto in house desenvolvido para a detecção sorológica contra B. anserina em G. gallus domesticus. Em média, os frangos infectados experimentalmente tornaram-se positivos aos 9º dia pós-inoculação (DPI), um valor do índice de densidade óptica (ODI) médio ± desvio padrão (SD) de 163,11 ± 70,65. O maior ODI observado foi 372,54 ± 132,39, em 26ºDPI, e o maior valor geral de ODI foi 626,51. Os frangos vacinados tornaram-se positivos entre 8º e 10° DPV, com um ODI de 245,59 a 10 DPV, com um ODI máximo geral de 543,13. Um total de 108 amostras de sangue foram coletadas de aves criadas em fazendas rústicas. Do total de amostras coletadas, 58,33% (63/108) foram consideradas positivas para B. anserina. O ODI máximo encontrado entre essas galinhas rústicas foi 283,24. Essa padronização proporcionou sensibilidade e especificidade de 100%.


Subject(s)
Animals , Poultry Diseases/diagnosis , Borrelia/immunology , Enzyme-Linked Immunosorbent Assay/veterinary , Serologic Tests/veterinary , Chickens/immunology , Antibodies/analysis
9.
Int. j. morphol ; 40(5): 1404-1414, 2022. ilus, tab
Article in English | LILACS-Express | LILACS | ID: biblio-1405270

ABSTRACT

SUMMARY: In Saudi Arabia, it is widely believed that women with reproductive problems can use the extract of the sage plant as a tea drink. This study was conducted to investigate the effects of this herb on the fertility of female rats and embryo implantation. Forty-eight Wistar virgin female rats were divided into four groups at random, with 12 rats in each group. The control group received distilled water orally. The three treatment groups received different concentrations of sage extract: 15, 60, or 100 mg/kg for 14 days before mating, then mated with a male and sacrificed on the 7th day of gestation, the uterine horns removed, and photographed. The total body weight of mothers, weight of uteri and ovaries and number of fetuses were determined. Ovarian and uteri tissues were cut into 5 µ sections and stained with hematoxylin and eosin. Serum FSH, LH were determined by the ELISA method. The present study showed that low dose of sage (15 mg/kg) have no effects on serum concentration levels of FSH and LH hormones, also has no effect on the number of growing follicles. The present study showed a significant differences (P≤0.05) in body weight, ovary and uterus weight in the groups treated with high doses of Salvia officinalis as compared to control group. Also a significant differences (P≤0.05) found in FSH, LH hormones. Histological study showed overall histomorphological structural configurations including growing and matured graafian follicular countable changes, besides a number of corpora lutea and regressed follicles in the treated groups with high doses of Salvia officinalis as compared to control group. The researchers concluded that the extract of the sage plant with high doses can stimulate the growth graafian follicles and improve fertility in female rats.


RESUMEN: En Arabia Saudita, se cree ampliamente que las mujeres con problemas reproductivos pueden usar el extracto de la planta de salvia como bebida de té. Este estudio se realizó para investigar los efectos de esta hierba sobre la fertilidad de las ratas hembra y la implantación del embrión. Se dividieron cuarenta y ocho ratas hembra vírgenes Wistar en cuatro grupos al azar, con 12 ratas en cada grupo. El grupo control recibió agua destilada por vía oral. Los tres grupos de tratamiento recibieron diferentes concentraciones de extracto de salvia: 15, 60 o 100 mg/kg durante 14 días antes del apareamiento, luego se aparearon con un macho y se sacrificaron el día 7 de gestación, se extrajeron los cuernos uterinos y se fotografiaron. Se determinó el peso corporal total de las madres, el peso del útero y los ovarios y el número de fetos. Los tejidos ováricos y uterinos se cortaron en secciones de 5 µ y se tiñeron con hematoxilina y eosina. FSH sérica, LH se determinaron por el método ELISA. El presente estudio mostró que dosis bajas de salvia (15 mg/kg) no tienen efectos sobre los niveles de concentración sérica de las hormonas FSH y LH, tampoco tienen efecto sobre el número de folículos en crecimiento. El presente estudio mostró diferencias significativas (P≤0,05) en el peso corporal, peso de ovario y útero en los grupos tratados con altas dosis de Salvia officinalis en comparación con el grupo control. También se encontraron diferencias significativas (P≤0,05) en las hormonas FSH, LH. El estudio histológico mostró configuraciones estructurales histomorfológicas generales que incluyen cambios contables en los folículos maduros (de Graaf) y en crecimiento, además de una cantidad de cuerpos lúteos y folículos en regresión en los grupos tratados con altas dosis de Salvia officinalis en comparación con el grupo de control. Los investigadores concluyeron que el extracto de la planta de salvia en altas dosis puede estimular el crecimiento de los folículos maduros y mejorar la fertilidad en ratas hembra.


Subject(s)
Animals , Female , Pregnancy , Rats , Embryo Implantation/drug effects , Plant Extracts/administration & dosage , Salvia officinalis/chemistry , Fertility/drug effects , Body Weight , Enzyme-Linked Immunosorbent Assay , Luteinizing Hormone/analysis , Administration, Oral , Follicle Stimulating Hormone/analysis
10.
Braz. J. Pharm. Sci. (Online) ; 58: e19660, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394027

ABSTRACT

Abstract In an attempt to increase molecular stability and provide controlled release, vascular endothelial growth factor (VEGF) was encapsulated into polycaprolactone (PCL) nanoparticles. Both VEGF-free and VEGF-loaded PCL nanoparticles were formulated by w/o/w double emulsion of the dichloromethane-water system in the presence of polyvinyl alcohol (PVA) and rat serum albumin. To achieve the optimal formulation concerning particle size and monodispersity, studies were carried out with different formulation parameters, including PVA concentration, homogenization time and rate. Scanning electron microscopy and dynamic light scattering analysis showed respectively that particles had a spherical shape with a smooth surface and particle size varying between 58.68-751.9 nm. All of the formulations were negatively charged according to zeta potential analysis. In vitro release study was performed in pH 7.4 phosphate-buffered saline at 37°C and released VEGF amount was measured by enzyme-linked immunosorbent assay (ELISA) method. At the end of the 35th day, 10% of total encapsulated VEGF was released with a sustained-release profile, which fitted the Korsmeyer-Peppas kinetic model. The bioactivation of the nanoparticles was evaluated using XTT and ELISA methods. As a result, the released VEGF was biologically active and also VEGF loaded PCL nanoparticles enhanced proliferation of the human umbilical vein endothelial cells in cell culture.


Subject(s)
Vascular Endothelial Growth Factor A , Nanoparticles/classification , In Vitro Techniques/methods , Enzyme-Linked Immunosorbent Assay/methods , Microscopy, Electron, Scanning/methods , Cell Culture Techniques/methods , Human Umbilical Vein Endothelial Cells
11.
Braz. J. Pharm. Sci. (Online) ; 58: e19791, 2022. tab, graf
Article in English | LILACS | ID: biblio-1383988

ABSTRACT

Abstract In China, Scutellaria is used for treating inflammatory-related diseases. Baicalin is the main active component of Scutellaria and has protective effects on acute pancreatitis. However, the mechanism of Baicalin is still unclear. In this study, the protective effects of baicalin on acute pancreatitis induced by taurocholate and its mechanism are investigated. In this study, mice were randomly divided into three groups: sham operation, model, and treatment groups. Acute pancreatitis in mice was induced by intraperitoneal injection of taurocholate (35 mg/kg). The treatment group was given baicalin (100 mg/kg) 2 h before acute pancreatitis induction. The mRNA expression levels of miR-429, nuclear factor kappa B65(NF-kB65), toll-like receptor 4(TLR4), TNF receptor associated factor6 (TRAF6), NF-kappa-B inhibitor(IkB), Follistatin-like 1 (FSTL1), and interleukin-1 receptor-associated kinase (IRAK) in the liver tissues 24 h after intraperitoneal injection were detected by RT-PCR. Then, the expression levels of NF-kB65, p-NF-κB65, TLR4, TRAF6, IkB, FSTL1, IRAK, p- IRAK, and p- IkB-а proteins were detected by Western blot. IL-6, TNF-α and IL-1 ß in plasma were measured by ELISA, and histopathological changes in the pancreases of the mice were observed. The results showed that after baicalin treatment, miR-429 expression in the pancreatic tissues and the expression levels of NF-kB65, TLR4, TRAF6, p-IkB-а, FSTL1, and p-IRAK decreased. Similarly, pancreatic myeloperoxidase (MPO) activity and the plasma levels of IL-6, TNF-а, IL-12, IL-1ß1, endotoxin, serum amylase, and lipase were reduced. Thus, the pancreatic injury induced by taurocholate was alleviated. The present study indicates that pretreatment with Baicalin can alleviate acute pancreatic injury induced by taurocholate in mice. The mechanism may be associated with the decreased miR-429 expression, reduced FSTL1 signaling pathway activity, TLR4 and TLR4/MyD88 signaling pathway inhibition, and reduced pancreatic inflammation. FSTL1 is the regulatory target for miR-429


Subject(s)
Animals , Male , Mice , HMGB1 Protein/adverse effects , Scutellaria/adverse effects , Injections/classification , Pancreatitis/pathology , Enzyme-Linked Immunosorbent Assay/instrumentation , Blotting, Western , Receptors, Tumor Necrosis Factor , Follistatin/administration & dosage , Liver/abnormalities
12.
Braz. J. Pharm. Sci. (Online) ; 58: e19332, 2022. tab, graf
Article in English | LILACS | ID: biblio-1384002

ABSTRACT

Chronic lymphocytic leukemia (CLL) is a blood cancer characterized by the accumulation of clonal B-lymphocytes. This study evaluated the mRNA gene expression of miR-15a, miR-16- 1, ZAP-70, and Ang-2 by qPCR, as well as the plasma levels of Bcl-2 by Elisa immunoassay, in CLL patients and healthy controls. Significant differences were observed when comparing patients and controls regarding miR-15a (p < 0.001), miR-16-1 (p < 0.001) mRNA, Ang-2 gene expression, and Bcl-2 plasma levels (p < 0.001). When stratified by risk, differences were maintained with a significantly reduced expression in high-risk patients. A positive correlation was observed between miR-15a and platelets (R2 = 0.340; p = 0.009) as well as between Bcl-2 and leukocytes (R2 = 0.310; p = 0.019). Conversely, negative correlations were observed between ZAP-70 and platelets (R2 = - 0.334; p = 0.011), between miR-15a and lymphocytes (R2 = - 0.376; p = 0.004), as well as between miR-16-and lymphocytes (R2 = - 0.515; p = 0.00004). The data suggest that a reduction in miR-15a and miR-16-1 expressions, in addition to an overexpression of Bcl-2, are associated with the reduction in apoptosis and, consequently, to a longer survival of lymphocytes, thus contributing to lymphocyte accumulation and aggravation of the disease. By contrast, Ang-2 expression was significantly higher in A than in B + C Binet groups. This context leads to the speculation that this biomarker should be investigated in more robust studies within populations with a still relevantly indolent form of the disease in an attempt to identify those patients with a greater potential for an aggravation of the disease


Subject(s)
Humans , Male , Female , Biomarkers/analysis , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , ZAP-70 Protein-Tyrosine Kinase/analysis , Patients , Enzyme-Linked Immunosorbent Assay/instrumentation , Gene Expression , Apoptosis
13.
Rev. chil. endocrinol. diabetes ; 15(1): 12-18, 2022. ilus, tab
Article in Spanish | LILACS | ID: biblio-1359333

ABSTRACT

El hiperaldosteronismo primario (HAP) es la causa más común de hipertensión arterial secundaria. A pesar de la prevalencia del HAP (6-10%) y sus consecuencias, los mecanismos que median los efectos deletéreos renales y extrarenales originados por la aldosterona más allá de la hipertensión arterial (ej. inflamación renal, alteraciones cardiacas y disfunción vascular), siguen siendo poco conocidos. Estudios previos sugieren que el exceso de aldosterona aumentaría proteínas sensibles a la activación del receptor de mineralocorticoides (MR), como las lipocalinas LCN2 (NGAL) y ORM1. OBJETIVO: Determinar la concentración de las lipocalinas ORM1, NGAL y NGAL-MMP9 en sujetos HAP. SUJETOS Y MÉTODOS: Estudio de cohorte transversal en sujetos adultos (similares en sexo, edad e IMC) separados en controles normotensos (CTL), hipertensos esenciales (HE) y con screening positivo de HAP (aldosterona ≥9 ng/dL y ARP < 1 ng/mL*h acorde a las guías internacionales de HAP). Se determinó la presión arterial sistólica (PAS) y diastólica (PAD), aldosterona plasmática, actividad renina plasmática (ARP) y la relación aldosterona / actividad de renina plasmática (ARR). Se determinó la concentración de NGAL, NGAL-MMP9 y ORM1 en suero por ELISA. RESULTADOS: Detectamos mayores niveles de ORM1 en sujetos HAP. No se detectaron diferencias en NGAL ni NGAL-MMP9 entre los grupos. Detectamos una asociación positiva de ORM1 con ARP (rho= -0,407, p=0,012) y con ARR (rho= 0,380 p= 0,021). CONCLUSIÓN: La mayor concentración de ORM1 en sujetos HAP y las asociaciones de ORM1 con aldosterona, ARP y ARR, proponen a esta proteína como un potencial biomarcador de HAP y de utilidad en el desarrollo de algoritmos diagnósticos de HAP.


Primary hyperaldosteronism (PA) is the most common cause of secondary hypertension. Despite the prevalence of PA (6-10%) and its consequences, the mechanisms that mediate the deleterious renal and extrarenal effects caused by aldosterone beyond arterial hypertension (eg renal inflammation, cardiac alterations and vascular dysfunction), remain barely known. Previous studies suggest that excess aldosterone would increase proteins sensitive to activation of the mineralocorticoid receptor (MR), such as lipocalins LCN2 (NGAL) and ORM1. AIM: To determine the concentration of the lipocalins ORM1, NGAL and NGAL-MMP9 in PA subjects. SUBJECTS AND METHODS: Cross-sectional study in adult subjects (similar in sex, age and BMI) grouped as normotensive controls (CTL), essential hypertensive (HE) and subjects with positive PA screening (aldosterone ≥ 9 ng/dL and PRA <1 ng/mL*h, according to international PA guidelines). Systolic (SBP) and diastolic (DBP) blood pressure, plasma aldosterone, plasma renin activity (PRA), and plasma aldosterone renin ratio (ARR) were determined. The concentration of NGAL, NGAL-MMP9 and ORM1 in serum was determined by ELISA. RESULTS: We detected higher levels Recibido: 03-09-2021 of ORM1 in PA subjects. No differences in NGAL or NGAL-MMP9 were detected between the groups. We detected a positive association of ORM1 with ARP (rho = -0.407, p < 0.05) and with ARR (rho = 0.380 p <0.05). CONCLUSION: The high levels of ORM1 in PA subjects and the associations of ORM1 with aldosterone, ARP and ARR, suggest ORM1 is a potential biomarker of PA, and useful in the development of a diagnostic algorithm for PA.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Young Adult , Orosomucoid/analysis , Biomarkers/blood , Lipocalins/analysis , Lipocalins/blood , Hyperaldosteronism/blood , Enzyme-Linked Immunosorbent Assay , Cross-Sectional Studies , Cohort Studies , Renin/analysis , Aldosterone/blood , Arterial Pressure , Hyperaldosteronism/diagnosis , Hypertension/diagnosis
14.
Braz. dent. sci ; 25(3): 1-5, 2022. tab
Article in English | LILACS, BBO | ID: biblio-1380063

ABSTRACT

Objective: Dental caries is one of the most common microbial diseases. Because of the infectious nature of the disease, the immunologic response by the host plays an essential role in its development. Therefore, the aim of this study was to evaluate the sCD14 levels in patients exhibiting two to three teeth with caries involving pulp along with apical periodontitis requiring root canal treatment. Material and Methods: This study was carried out on 20 participants, of whom 10 were caries-free (Control) and 10 had two to three teeth with symptomatic irreversible pulpitis along with apical periodontitis requiring root canal treatment, within the ages of 20- 30 years. Unstimulated saliva of the participants was collected with disposable needle-less syringe from buccal and labial vestibules. The sCD14 levels in salivary samples were assessed before and following endodontic treatment. The results were analyzed by ELISA. Results: The obtained levels of sCD14 were analyzed statistically. Paired T test was performed to assess the significance. The results revealed that there was a significant difference in sCD14 levels with a P=0.0005, as it had drastically reduced once the inflammation has subsided. Conclusion: Higher values of sCD14 levels were seen in patients with symptomatic irreversible pulpitis along with apical periodontitis than in caries free group. The study also showed that sCD levels were significantly reduced following post endodontic treatment. Therefore, increased levels of sCD14 can be considered as a marker of inflammation. (AU)


Objetivo: A cárie dentária é uma das doenças microbianas mais comuns. Devido à natureza infecciosa da doença, a resposta imunológica do hospedeiro desempenha um papel essencial no seu desenvolvimento. Portanto, o objetivo deste estudo foi avaliar os níveis de sCD14 em pacientes que possuiam dois a três dentes com necessidade de tratamento endodôntico por apresentarem lesão de cárie envolvendo polpa e periodontite periapical. Material e Métodos: Este estudo foi realizado em 20 participantes, dos quais 10 estavam livres de cárie (controle) e 10 tinham dois a três dentes com pulpite irreversível sintomática e periodontite periapical com necessidade de tratamento endodôntico, nas idades de 20 a 30 anos. A saliva não estimulada das crianças foi coletada com seringa descartável sem agulha dos vestíbulos bucal e labial. Os níveis de sCD14 em amostras salivares foram avaliados antes e após o tratamento endodôntico. Os resultados foram analisados por ELISA. Resultados: Os níveis de sCD14 obtidos foram analisados estatisticamente. O teste T pareado foi realizado para avaliar a significância. Os resultados revelaram que houve uma diferença significativa nos níveis de sCD14 com um P = 0,0005, uma vez que reduziu drasticamente uma vez que a inflamação diminuiu. Resultados: Os níveis de sCD14 obtidos foram analisados estatisticamente. O teste T pareado foi realizado para avaliar a significância. Os resultados revelaram que houve uma diferença significativa nos níveis de sCD14 com um P = 0,0005, uma vez que reduziu drasticamente uma vez que a inflamação diminuiu. Conclusão: Valores mais elevados de níveis de sCD14 foram observados em pacientes com pulpite irreversível sintomática junto com periodontite periapical do que no grupo livre de cárie. O estudo também mostrou que os níveis de sCD foram significativamente reduzidos após o tratamento endodôntico. Portanto, níveis aumentados de sCD14 podem ser considerados um marcador de inflamação. (AU)


Subject(s)
Humans , Adult , Periapical Periodontitis , Enzyme-Linked Immunosorbent Assay , Lipopolysaccharide Receptors , Dental Caries
15.
Evid. actual. práct. ambul ; 25(2): e007014, 2022. ilus, tab
Article in Spanish | LILACS, BINACIS, UNISALUD | ID: biblio-1380221

ABSTRACT

El nuevo tratamiento simplificado con antivirales orales para pacientes con Hepatitis C puede ser abordado desde la atención primaria, lo que facilita el acceso de la población afectada por esta infección crónica. En este artículo se repasan los aspectos claves del diagnóstico, el esquema de tratamiento simplificado y los candidatos a recibirlo. (AU)


The new simplified treatment with oral antivirals for hepatitis C patients can be approached at the primary care level, facilitating access for the population affected by this chronic infection. This article reviews the key aspects of the diagnosis, the simplified treatment scheme, and the eligible candidates for the treatment. (AU)


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Antiviral Agents/administration & dosage , Hepatitis C/diagnosis , Hepatitis C/drug therapy , Primary Health Care , Enzyme-Linked Immunosorbent Assay , Hepatitis C/blood , Persistent Infection/diagnosis , Persistent Infection/drug therapy , Persistent Infection/blood , Liver Cirrhosis/diagnosis
16.
Article in Chinese | WPRIM | ID: wpr-939704

ABSTRACT

OBJECTIVE@#To establish a new method for synthesizing Lewis blood group antigens, that is, the mimotopes of Lewis blood group antigens were screened by using an alpaca phage display nanobody library.@*METHODS@#We selected mimotopes of the Lewis a (lea) antigen by affinity panning of an alpaca phage display nanobody library using a monoclonal anti-lea antibody. Enzyme-linked immunosorbent assay (ELISA) was used to test the affinity of the positive clones for the monoclonal anti-lea antibody, and the high-affinity positive clones were selected for sequencing and synthesis. Finally, the sensitivity, specificity and reactivity of the synthesized lea mimotope in clinical samples were verified by ELISA.@*RESULTS@#A total of 96 phage clones were randomly selected, and 24 were positive. Fourteen positive clones with the highest affinity were selected for sequencing. The result showed that there were 5 different sequences, among which 3 sequences with the highest frequency, largest difference and highest affinity were selected for expression and synthesis. The sensitivity and specificity of lea mimic antigen by ELISA showed that, the minimum detection limit of gel microcolumn assay (GMA) and ELISA method were 25 times different, and the lea mimic antigen had no cross reacted with the other five unrelated monoclonal antibodies(P<0.001). Finally, 30 clinical plasma samples were analyzed. The mean absorbance of the 15 positive plasma samples was significantly higher than that of the 15 negative plasma samples (P=0.02). However, the positive signal values of the clinical samples were much lower than those of the monoclonal antibodies.@*CONCLUSION@#A new method of screening lea mimic antigen by using alpaca phage nanoantibody library has been established, which is expected to realize the screening of lea mimotopes, thus realizing the application of high-sensitivity detection methods such as ELISA and chemiluminescence in blood group antibody identification.


Subject(s)
Animals , Antibodies, Monoclonal , Antineoplastic Agents, Immunological , Bacteriophages , Blood Group Antigens , Camelids, New World , Enzyme-Linked Immunosorbent Assay/methods , Epitopes , Humans , Lewis Blood Group Antigens , Peptide Library
17.
Article in Chinese | WPRIM | ID: wpr-935240

ABSTRACT

Objective: To express DNA-binding protein (DBP) of human adenovirus (HAdV) type 7 using the prokaryotic expression system, and product anti-HAdV-7 DBP rabbit polyclonal antibody. Methods: The HAdV-7 DBP gene was synthesized and cloned into prokaryotic expressing vector pET30a, and the recombinant plasmid was transformed into E. coli BL21 (DE3) competent cell. The recombinant protein DBP was expressed by induced Isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified with Ni-NTA affinity column. The titer of anti-DBP polyclonal antibody produced in immunized rabbit was measured by indirect ELISA, and the specificity of the antibody was identified by Western blotting and indirect immunofluorescence assay (IFA). In addition, purified rDBP was used as coating antigen for indirect ELISA assay to detect specific IgM and IgG antibodies against DBP in the serum of children infected with HAdV. Results: The HAdV-7 DBP plasmid was constructed successfully. The purified recombinant DBP was more than 95% after purification. The titer of polyclonal antibody was 1∶1 024 000. The polyclonal antibody showed high specificity in vitro using Western blotting and IFA. The positive rate of specific anti-DBP IgM and IgG antibody in acute-phase serum samples collected from children infected with HAdV were 50.0% (19/38) and 63.2% (24/38), respectively, using indirect ELISA. Conclusion: In summary, the HAdV-7 rDBP is expressed using prokaryotic expression system, and the recombinant HAdV-7 DBP protein and the anti-DBP rabbit polyclonal antibody with high titer are prepared.


Subject(s)
Adenoviruses, Human/genetics , Animals , Antibody Specificity , Blotting, Western , DNA-Binding Proteins/metabolism , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Immunoglobulin G , Rabbits
18.
Article in Chinese | WPRIM | ID: wpr-928742

ABSTRACT

OBJECTIVE@#To evaluate the value of high mobility group protein B1 (HMGB1) and soluble receptor for advanced glycation end products (sRAGE) in the diagnosis, efficacy monitoring and prognosis of newly diagnosed multiple myeloma (MM) patients.@*METHODS@#Fifty newly diagnosed MM patients before and after chemotherapy and 50 hematological outpatients from October 2018 to May 2020 were selected. Enzyme linked immunosorbent assay (ELISA) was used to detect the serum HMGB1 and sRAGE levels of the patients. ROC was used to further analyze the efficacy of serum HMGB1 and sRAGE levels on the diagnosis of MM. At the same time, the serum levels of HMGB1 and sRAGE before and after chemotherapy were compared, and their values in the evaluation of curative effect of MM patients were analyzed. According to the mean values of serum HMGB1 and sRAGE, all the patients were divided into different groups, the clinical characteristics and survival status of the patients were compared.@*RESULTS@#Before treatment the serum HMGB1 level of the patients in MM group was higher than that in control group, while sRAGE level was lower (t=11.363,6.127, P<0.001). The AUC of serum HMGB1 and sRAGE in the MM patients was 0.955 and 0.811, respectively. After 3 courses of chemotherapy, HMGB1 level of the patients in CR group was lower than before chemotherapy, while in PD group was higher, as well as sRAGE level of the patients in PR group (P<0.05). There were significant differences in R-ISS stage, HGB, CRP, ESR, CD56, CD117, D13S319 deletion between HMGB1 high expression group and HMGB1 low expression group (χ2=3.920, 6.522, 6.65, 4.16, 3.945, 6.65, 4.16, P<0.05), while there were significant differences in ISS stage, CRP and CD56 between sRAGE low expression group (28 cases) and sRAGE high expression group (22 cases) (χ2=4.565, 4.711, 5.547, P<0.05). Kaplan-Meier survival analysis showed that the patients in HMGB1 low expression group had better survival condition, for PFS Tlow>Thigh (χ2=9.470, P<0.05), and for OS Tlow>Thigh (χ2=7.808, P<0.05); there was no difference in the survival of sRAGE high expression group and low expression group, for PFS Tlow<Thigh (χ2=1.661, P>0.05), and for OS Tlow<Thigh (χ2=2.048, P>0.05). Cox analysis showed that LDH and HMGB1 were the factors affecting the prognosis of the patients, and both of them affected PFS (HR=2.771, 95% CI: 1.002-7.662, P=0.049; HR=6.022, 95% CI: 1.689-21.470, P=0.006), while HMGB1 also affected OS (HR=4.275, 95% CI: 1.183-15.451, P=0.027).@*CONCLUSION@#The serum HMGB1 and sRAGE have certain auxiliary value for the diagnosis and curative effect monitoring of newly diagnosed MM patients, and serum HMGB1 is expected to be an auxiliary detection index for the prognosis of MM.


Subject(s)
Enzyme-Linked Immunosorbent Assay , HMGB1 Protein/blood , Humans , Multiple Myeloma/therapy , Prognosis , Receptor for Advanced Glycation End Products/blood
19.
Article in Chinese | WPRIM | ID: wpr-927910

ABSTRACT

Puerarin was conjugated with bovine serum albumin(BSA) and ovalbumin(OVA) by periodate oxidation to serve as the immunogen and coating antigen, respectively. BALB/c mice were immunized with puerarin-BSA according to the routine immunization procedure, and the titer and specificity of serum were detected after three immunization. After booster immunization, mouse spleen lymphocytes were fused with mouse myeloma cells, and 24 hybridoma cell lines of the monoclonal antibodies against puerarin were screened by monoclonal antibody screening technique. Ascites was prepared and purified. The cross-reactivity of monoclonal antibody(mAb) M1 with 4'-methoxy puerarin, daidzin, puerarin-6″-O-xyloside, daidzein, mirificin, 3'-methoxy puerarin, and 3'-hydroxy puerarin was 239.84%, 112.18%, 67.89%, 58.28%, 22.37%, 0.40%, and 0.20%, respectively, and those with other analogs such as baicalein and baicalin were all less than 0.10%. The IC_(50) and the working range of the indirect competitive enzyme-linked immunosorbent assay(icELISA) for puerarin were 44.80 ng·mL~(-1) and 8.20-292.30 ng·mL~(-1), respectively. The average recovery was 91.95%-98.20% with an RSD in the range of 0.70%-2.60%. The content of puerarin in different Puerariae Lobatae Radix samples was determined with icELISA and validated by UPLC-MS. The correlation between data obtained from icELISA and UPLC-MS was 0.999 0, indicating that icELISA is suitable for the rapid detection of puerarin in Puerariae Lobatae Radix samples.


Subject(s)
Animals , Antibodies, Monoclonal , Chromatography, Liquid , Enzyme-Linked Immunosorbent Assay/methods , Hybridomas/metabolism , Isoflavones , Mice , Mice, Inbred BALB C , Tandem Mass Spectrometry
20.
Chinese Journal of Biotechnology ; (12): 185-195, 2022.
Article in Chinese | WPRIM | ID: wpr-927703

ABSTRACT

Clostridium difficile is an important zoonotic intestinal pathogen, which is widely present in humans and a variety of animals. The ST11 type C. difficile is one of the most widespread and harmful subtypes in the world. As a large country in pig farming, China lacks efficient methods for detecting C. difficile of porcine origin, leaving hidden dangers for the prevention and control of C. difficile. The aim of this study was to develop a specific and sensitive double-antibody sandwich ELISA for the epidemiological investigation of ST11 type C. difficile of porcine origin. Firstly, a 97 kDa receptor binding domain (RBD) was expressed in a prokaryotic host and purified. A hybridoma cell line AE2D3 capable of stably secreting monoclonal antibody targeting the RBD was screened, and the antibody subtype was determined to be IgG2b (κ). Secondly, a double antibody sandwich ELISA method was developed, where the monoclonal antibody targeting the RBD was used as a detection antibody, and the rabbit polyclonal antibody was used as a capture antibody. The chessboard method was used to determine the matching concentration of the capture antibody and the detection antibody, the antigen coating conditions, the blocking conditions, the incubation conditions for detection antibody and samples to be tested, as well as the reaction conditions of HRP-conjugated and reaction conditions of TMB chromogenic solution. The negative cutoff OD450 was 0.152, and no cross-reaction with 13 strains of non-ST11 type C. difficile was found. The minimum detection concentration of RBD was 8.83 ng/mL. This specific and sensitive double-antibody sandwich ELISA provides a reliable serological detection method for epidemiological investigation of the ST11 type C. difficile in pig industry.


Subject(s)
Animals , Antibodies, Monoclonal , Bacterial Proteins/genetics , Bacterial Toxins , Clostridioides difficile , Enzyme-Linked Immunosorbent Assay , Hybridomas , Swine
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