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Electron. j. biotechnol ; 51: 40-49, May. 2021. tab, ilus, graf
Article in English | LILACS | ID: biblio-1343322


BACKGROUND: Scavenger receptor class B (SRB) is a multifunctional protein in animals that participates in physiological processes, including recognition of a wide range of ligands. Astaxanthin is a major carotenoid found in shrimp. However, the molecular mechanism of astaxanthin and SRB protein binding has not been reported. RESULTS: In the present study, a member of the SRB subfamily, named PmSRB, was identified from the transcriptome of black tiger shrimp (Penaeus monodon). The open reading frame of PmSRB was 1557 bp in length and encoded 518 amino acids. The structure of PmSRB included a putative transmembrane structure at the N-terminal region and a CD36 domain. Multiple sequence alignment indicated that the CD36 domain were conserved. Phylogenetic analysis showed four separate branches (SRA, SRB, SRC, and croquemort) in the phylogenetic tree and that PmSRB was clustered with SRB of Eriocheir sinensis. Quantitative real-time polymerase chain reaction showed that the PmSRB gene was widely expressed in all tissues tested, with the highest expression level observed in the lymphoid organ and brain. Subcellular localization analysis revealed that PmSRB-GFP (green fluorescent protein) fusion proteins were predominantly localized in the cell membrane. The recombinant proteins of PmSRB showed binding activities against astaxanthin in vitro. CONCLUSIONS: PmSRB was identified and characterized in this study. It is firstly reported that PmSRB may take as an important mediator of astaxanthin uptake in shrimp.

Animals , Penaeidae , Receptors, Scavenger/metabolism , In Vitro Techniques , Blotting, Western , Chromatography, High Pressure Liquid , Sequence Alignment , Xanthophylls , Receptors, Scavenger/isolation & purification , Receptors, Scavenger/genetics , Real-Time Polymerase Chain Reaction/methods , Transcriptome
Rev. invest. clín ; 73(2): 120-126, Mar.-Apr. 2021. graf
Article in English | LILACS | ID: biblio-1251872


ABSTRACT Background: Underestimation of the number of cases during the coronavirus disease 2019 (COVID-19) pandemic has been a constant concern worldwide. Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA using real-time reverse-transcription polymerase chain reaction (RT-PCR) is the most common method to confirm a case. However, these tests have suboptimal sensitivity. Objective: The objective of the study was to estimate the number of COVID-19 confirmed cases, intensive care unit (ICU) admissions and deaths in Mexico, accounting for the probabilities of false-negative tests. Methods: We used publicly available, national databases of all SARS-CoV-2 tests performed at public laboratories in Mexico between February 27 and October 31, 2020. We used the estimated probabilities of false-negative tests based on the day of clinical sample collection after symptom initiation calculated previously. With the resulting model, we estimated the corrected daily number of cases, ICU admissions, and deaths. Results: Among 2,024,822 people tested in Mexico between February 27 and October 31 with an available result, we estimated 1,248,583 (95% confidence interval 1,094,850-1,572,818) cases, compared to 902,343 cases reported with positive tests. ICU admissions and deaths were 15% and 8% higher than reported, respectively. Conclusion: Accounting for SARS-CoV-2 RT-PCR-based diagnostic testsҠprecision is a simple way to improve estimations for the true number of COVID-19 cases among tested persons.

Humans , COVID-19 Testing/methods , COVID-19/diagnosis , Databases, Factual , Sensitivity and Specificity , Reverse Transcriptase Polymerase Chain Reaction , False Negative Reactions , Real-Time Polymerase Chain Reaction/methods , COVID-19/mortality , COVID-19/epidemiology , Hospitalization/statistics & numerical data , Intensive Care Units/statistics & numerical data , Mexico/epidemiology
Electron. j. biotechnol ; 50: 53-58, Mar. 2021. graf, tab, ilus
Article in English | LILACS | ID: biblio-1292393


BACKGROUND: Lycium barbarum (also called wolfberry), a famous Chinese traditional medicine and food ingredient, is well recognized for its significant role in preventing obesity; however, the molecular mechanisms underlying its preventive effects on fat accumulation are not well understood yet. The aim of this study was to determine the effects and mechanism of Lycium barbarum polysaccharides (LBP) on the proliferation and differentiation of 3T3-L1 preadipocytes. MTT was used to detect the proliferation of 3T3-Ll preadipocytes. Oil red O staining and colorimetric analysis were used to detect cytosolic lipid accumulation during 3T3-L1 preadipocyte differentiation. Real-time fluorescent quantitative PCR (qPCR) technology was used to detect peroxisome proliferator-activated receptor c (PPARc), CCAAT/enhancer-binding protein a (C/EBPa), adipocyte fatty-acid-binding protein (aP2), fatty acid synthase (FAS), and lipoprotein lipase (LPL) expression. RESULTS: The concentration of LBP from 25 to 200 lg/mL showed a tendency to inhibit the growth of preadipocytes at 24 h, and it inhibited the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner. In the preadipocytes treated with 200 lg/mL LBP, there were reduced lipid droplets in the cytoplasm, and its effect was opposite to that of rosiglitazone (ROS), which significantly reduced the PPARc, C/EBPa, aP2, FAS, and LPL mRNA expression of adipocytes. CONCLUSIONS: LBP exerts inhibitive effects on the proliferation and differentiation of 3T3-L1 preadipocytes and decreases the cytoplasm accumulation of lipid droplets during induced differentiation of preadipocytes toward mature cells. Above phenomenon might link to lowered expression of PPARc, C/EBPa, aP2, FAS, and LPL after LBP treatment. Thus, LBP could serve as a potential plant extract to treat human obesity or improve farm animal carcass quality via adjusting lipid metabolism.

Polysaccharides , Plant Extracts , Adipocytes , Lycium/chemistry , Cell Differentiation , 3T3-L1 Cells , Cell Proliferation , Adipogenesis , Real-Time Polymerase Chain Reaction/methods
Electron J Biotechnol ; 49: 34-41, Jan. 2021. tab, graf
Article in English | LILACS | ID: biblio-1291638


BACKGROUND: This work studied how the exposure to an unusual substrate forced a change in microbial populations during anaerobic fermentation of crude glycerol, a by-product of biodiesel production, with freshwater sediment used as an inoculum. RESULTS: The microbial associations almost completely (99.9%) utilized the glycerol contained in crude glycerol 6 g L 1 within four days, releasing gases, organic acids (acetic, butyric) and alcohols (ethanol, n-butanol) under anaerobic conditions. In comparison with control medium without glycerol, adding crude glycerol to the medium increased the amount of ethanol and n-butanol production and it was not significantly affected by incubation temperature (28 C or 37 C), nor incubation time (4 or 8 d), but it resulted in reduced amount of butyric acid. Higher volume of gas was produced at 37 C despite the fact that the overall bacterial count was smaller than the one measured at 20 C. Main microbial phyla of the inoculum were Actinobacteria, Proteobacteria and Firmicutes. During fermentation, significant changes were observed and Firmicutes, especially Clostridium spp., began to dominate, and the number of Actinobacteria and Gammaproteobacteria decreased accordingly. Concentration of Archaea decreased, especially in medium with crude glycerol. These changes were confirmed both by culturing and culture-independent (concentration of 16S rDNA) methods. CONCLUSIONS: Crude glycerol led to the adaptation of freshwater sediment microbial populations to this substrate. Changes of microbial community were a result of a community adaptation to a new source of carbon.

Bacteria/isolation & purification , Geologic Sediments/microbiology , Fresh Water/microbiology , Glycerol/metabolism , Bacteria/metabolism , Adaptation, Biological , Biofuels , Fermentation , Real-Time Polymerase Chain Reaction/methods , Anaerobiosis
J. venom. anim. toxins incl. trop. dis ; 27: e20200118, 2021. tab, graf, mapas
Article in English | LILACS, VETINDEX | ID: biblio-1154768


The early symptoms of leptospirosis and dengue fever are difficult to distinguish and can cause diagnostic confusion. Due to the large dengue epidemics that has occurred in Brazil in recent years, it is possible that cases of leptospirosis were unreported. Therefore, we performed a retrospective study to detect leptospirosis in patients who were tested for dengue, but whose laboratory diagnoses were negative. Methods: Sera samples from 2,017 patients from 48 cities located in the central region of São Paulo state, Brazil, were studied. All samples were subjected to the microscopic agglutination test (MAT), 305 of which were taken from patients five days or less since the onset of symptoms, and were additionally subjected to real-time polymerase chain reaction (PCR). Results: The overall prevalence of leptospirosis cases was 21 (1.04%), with 20 through MAT (18 for Icterohaemorrhagiae and two for the Cynopteri serogroup) and one through PCR (amplicon sequencing compatible with Leptospira interrogans). According to previously established criteria, eight cases of leptospirosis were classified as "confirmed" and 13 as "probable". The Brazilian notification system for health surveillance had no records for 16 patients positive for leptospirosis and, thus, they were considered unreported cases. Statistical analyses revealed that the prevalence of leptospirosis was higher in men (1.56%) than in women (0.56%), and the mean age was higher in positive patients (43.7 years) than in negative ones (32.3 years). Conclusion: The results indicated that patients suspected of dengue fever had evidence of leptospirosis or Leptospira infection, and most of these cases were unreported in the Brazilian notification system. The high burden of dengue may contribute to the misdiagnosis of leptospirosis, and health professionals should increase their awareness of leptospirosis as an important differential diagnosis of patients with suspicion of dengue.(AU)

Humans , Dengue/diagnosis , Real-Time Polymerase Chain Reaction/methods , Leptospirosis/diagnosis , Health Surveillance , Agglutination Tests
Rev. Hosp. Ital. B. Aires (2004) ; 40(3): 117-125, sept. 2020. ilus, tab
Article in Spanish | LILACS | ID: biblio-1129078


En diciembre de 2019 se identificó el virus SARS-CoV-2, cuya rápida propagación global puso en estado de emergencia al mundo entero, llevando al ser humano a una situación sin antecedente cercano. El objetivo de esta revisión es describir los métodos diagnósticos utilizados actualmente para identificar la infección por SARS-CoV-2. Las manifestaciones clínicas y el espectro imagenológico de la enfermedad son muy inespecíficos y no permiten realizar un diagnóstico certero. Por esta razón, es esencial una apropiada toma de muestra respiratoria en el momento y sitio anatómico adecuado para un diagnóstico preciso de COVID-19. La técnica de muestreo más utilizada es el hisopado nasofaríngeo y la prueba diagnóstica más fiable se basa en la retrotranscripción seguida por reacción en cadena de la polimerasa en tiempo real (RT-PCR). No obstante, existen otras técnicas moleculares, como también tests serológicos para detectar anticuerpos o fragmentos antigénicos del SARS-CoV-2. Más allá de la precisión diagnóstica, es importante tener en cuenta la probabilidad basal (pretest) para interpretar correctamente el resultado obtenido y aislar aquellos posibles falsos negativos. Con el objetivo de evitar la saturación del sistema de salud es imprescindible contar con información y métodos diagnósticos precisos para detectar tempranamente los focos de infección y reducir la transmisión comunitaria, utilizando eficazmente los diferentes recursos diagnósticos. (AU)

In December 2019, the SARS-CoV-2 virus was identified for the first time, whose rapid global spread put the entire world in a state of emergency, leading humans to an unprecedented situation with no immediate history. The main purpose of this review is to describe the diagnostic methods currently used to identify SARS-CoV-2 infection. The clinical manifestations and the imaging spectrum of the disease are nonspecific and do not allow an accurate diagnosis to be made. For this reason, an appropriate respiratory sampling at the right time and anatomical site is essential for an accurate diagnosis of COVID-19. The most widely used sampling technique is nasopharyngeal swab, and the most reliable diagnostic test is by reverse transcription followed by real-time polymerase chain reaction (RT-PCR). However, there are other molecular techniques, as well as serological tests to detect antibodies or antigenic fragments of SARS-CoV-2. Beyond the diagnostic precision, it is important to take into account the baseline probability (pre-test) to correctly interpret the result obtained and isolate those possible false negatives. In order to avoid saturation of the health system, it is essential to have accurate information and diagnostic methods to detect outbreaks of infection in early stages and to reduce communitary transmission, making effective use of the various diagnostic resources. Coronavirus infections/diagnosis, viral/diagnosis, pandemics, clinical laboratory techniques, real-time polymerase chain reaction, antigens, viral/analysis. (AU)

Humans , Serologic Tests/methods , Coronavirus Infections/diagnosis , Real-Time Polymerase Chain Reaction/methods , Argentina , Pneumonia, Viral/diagnosis , Serologic Tests/statistics & numerical data , Polymerase Chain Reaction/methods , Polymerase Chain Reaction/statistics & numerical data , Coronavirus Infections/physiopathology , Coronavirus Infections/prevention & control , Coronavirus Infections/diagnostic imaging , False Negative Reactions , False Positive Reactions , Real-Time Polymerase Chain Reaction/statistics & numerical data , Betacoronavirus
Rev. cuba. med. trop ; 72(2): e522, mayo.-ago. 2020. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1149912


Introducción: Los ensayos para cuantificar el ADN del virus de la hepatitis B (VHB) o carga viral son imprescindibles en el diagnóstico y en el seguimiento de los pacientes con hepatitis B crónica; de ahí que estén disponibles estuches diagnósticos para esta función. En el presente estudio se muestra la validación de SUMASIGNAL VHB (un paso), el cual es un sistema de reacción en cadena de la polimerasa en tiempo real (RCP-TR) para la cuantificación del genoma del VHB, propuesto por el Centro de Inmunoensayo. Objetivo: Evaluar el desempeño analítico de SUMASIGNAL VHB (un paso). Métodos: Se utilizó un panel de 80 muestras de suero bien caracterizadas y el Tercer Estándar Internacional de la OMS para las técnicas de amplificación de ácidos nucleicos del virus de la hepatitis B. Se determinaron las características del ensayo como especificidad clínica, especificidad analítica (reactividad cruzada), rango lineal o linealidad y exactitud, precisión intraensayo y comparación con un ensayo de referencia. Resultados: La especificidad analítica y clínica fue del 100 por ciento. Al evaluar la linealidad y exactitud con un estándar de referencia de la OMS, se obtuvo que la totalidad de las diferencias entre los Log10 del valor obtenido y el de referencia resultaron inferiores a 0,5 Log10 (r= 0,9977 y r2= 0,9954). Además, se obtuvieron bajos coeficientes de variación intraensayo. La evaluación comparativa con el estuche comercial Artus HBV RG PCR kit mostró una correlación fuerte (r= 0,8882). Conclusiones: SUMASIGNAL VHB (un paso) es un ensayo fácil de realizar manualmente, es rápido e incluye reactivos de extracción de ácidos nucleicos. Teniendo en cuenta la validez del método para el uso previsto, puede ser recomendado para su introducción en el diagnóstico, la vigilancia y la indicación de tratamiento en los pacientes con hepatitis B crónica(AU)

Introduction: Assays to quantify hepatitis B virus (HBV) DNA or viral load are indispensable for the diagnosis and follow-up of patients with chronic hepatitis B, hence the availability of diagnostic kits for this purpose. The present study deals with the validation of HBV SUMASIGNAL (one step), a real time polymerase chain reaction (RT-PCR) system for quantification of the HBV genome proposed by the Immunoassay Center. Objective: Evaluate the analytical performance of HBV SUMASIGNAL (one step). Methods: Use was made of a panel of 80 well characterized serum samples and the Third WHO International Standard for hepatitis B virus nucleic acid amplification techniques. Determination was performed of assay characteristics such as clinical specificity, analytical specificity (cross-reactivity), linear range or linearity and accuracy, intra-assay precision and comparison with a reference assay. Results: Analytical and clinical specificity was 100 percent. Evaluation of linearity and accuracy with a WHO reference standard revealed that all the differences between the log10 of the value obtained and the reference value were lower than 0.5 log10 (r= 0.9977 and r2= 0.9954). The intra-assay variation coefficients obtained were low. Comparative evaluation with the commercial Artus HBV RG PCR kit showed a strong correlation (r= 0.8882). Conclusions: The assay HBV SUMASIGNAL (one step) is easy to conduct manually, fast and includes reagents for nucleic acid extraction. Based on the validity of the method for the use in mind, it may be recommended for incorporation into the diagnosis, surveillance and treatment of patients with chronic hepatitis B(AU)

Humans , Hepatitis B virus/isolation & purification , Nucleic Acid Amplification Techniques/methods , Real-Time Polymerase Chain Reaction/methods , Validation Study
Vaccimonitor (La Habana, Print) ; 29(2)mayo.-ago. 2020. tab, graf
Article in English | LILACS, CUMED | ID: biblio-1127512


The objective of this study was to investigate the effects of Spirulina platensis (SP) powder supplementation on immune response in SPF chickens. For this purpose, 120 SPF chicks were randomly clustered into six groups consisting of 20 birds each which assigned to five groups vaccinated by commercial inactivated Newcastle disease (ND) vaccine at 21 days of age. The four groups were supplemented with 0.5, 1, 1.5 and 2 g of SP per kg of ration at 7 day of age and other group as control treatment group. Control unvaccinated group still without any treatment. Individual blood samples were collected weekly from all groups, and NDV-HI antibodies were measured using Hemagglutination inhibition (HI) test. After 28 days post-vaccination, ten birds from all groups were challenged intramuscularly at a dose 0.5 mL/bird containing 106 EID50 of local NDV genotype VII. Challenge virus shedding was detected using real time qrt-PCR of oropharyngeal swabs that were collected from all challenged chicken groups of at 3, 5, 7 and 10 days post challenge. Obtained results showed that vaccinated groups of SPF-chickens either supplied with Spirulina or control treatment group induced positive serological response as NDV-HI antibody were measured in sera of immunized chicks (7.6, 8, 8.3, 8.9 and 7.4 log2, respectively) at 4 weeks post vaccination (WPV). Significant differences were observed at 2 WPV in the vaccinated SPF chickens consumed 1, 1.5 and 2 g of SP/kg of ration, compared to untreated vaccinated group (p<0.05). Immunized SPF chickens supplied with different SP concentration confer satisfactory protection against heterologous challenge virus (90 percent, 100 percent, 100 percent and 100 percent respectively), in contrast to untreated vaccinated chickens. Different percentages of reduction of viral shedding (55 percent, 65 percent, 76 percent and 87 percent) of treated vaccinated chickens with different concentration of SP were detected, despite untreated group were reduced 46 percent from total viral shedding. These findings suggest that dietary Spirulina has immune-stimulatory effects on the immune system of SPF chickens. One gram from SP per kg of ration was minimum recommended concentration that able to exhibit optimum immune response, increase protection against heterologous strains and able to reduce viral shedding(AU)

El objetivo de este estudio fue investigar los efectos de la suplementación con polvo de Spirulina platensis (SP) sobre la respuesta inmune en pollos SPF. Para este propósito se agruparon al azar 120 polluelos SPF en seis grupos de 20 aves cada uno, que se asignaron a cinco grupos vacunados con la vacuna comercial inactivada contra la enfermedad de Newcastle (ND) a los 21 días de edad. Cuatro grupos se suplementaron con 0,5; 1; 1,5 y 2 g de SP por kg de ración a los 7 días de edad, un grupo vacunado sin suplemento y un grupo sin ningún tratamiento. Semanalmente, se recogieron muestras de sangre individuales de todos los grupos y se midieron los anticuerpos hemaglutinantes contra el virus Newcastle (NDV-HI) mediante la prueba de inhibición de la hemaglutinación (HI). 28 días después de la vacunación, fueron retadas diez aves de cada grupo por vía intramuscular a una dosis 106 EID50 del genotipo VII del NDV local en un volumen de 0,5 mL/ave. Se detectó la eliminación del virus mediante qrt-PCR en hisopos orofaríngeos que se recolectaron en todos los grupos a los 3, 5, 7 y 10 días después del reto. Los resultados obtenidos mostraron que los grupos vacunados de pollos y suplementados con Espirulina y el grupo de control vacunado, indujeron una respuesta serológica positiva cuando se determinaron los anticuerpos NDV-HI en los pollitos inmunizados (7,6; 8; 8,3; 8,9 y 7,4 log2 respectivamente) a las 4 semanas después de la vacunación (SPV). Se observaron diferencias significativas a las 2 SPV en los pollos vacunados que consumieron 1, 1,5 y 2 g de SP/kg de ración, en comparación con el grupo vacunado no tratado (p<0,05). Los pollos inmunizados que recibieron diferentes concentraciones de SP mostraron una protección satisfactoria contra el desafío heterólogo viral (90 por ciento, 100 por ciento y 100 por ciento respectivamente), en contraste con los pollos vacunados no tratados. Se observaron diferentes porcentajes de reducción de la diseminación viral (55 por ciento, 76 por ciento y 87 por ciento) entre los pollos vacunados tratados con diferente concentración de SP. En el grupo no tratado se redujo al 46 por ciento. Estos hallazgos sugieren que la Espirulina en la dieta tiene efectos inmunoestimuladores sobre el sistema inmunitario de los pollos. Un gramo de SP por kg de ración fue la concentración mínima recomendada para una respuesta inmune óptima, y de esta forma aumentar la protección contra las cepas heterólogas y disminuir la diseminación viral(AU)

Humans , Male , Female , Newcastle disease virus/pathogenicity , Vaccines, Inactivated , Chickens , Spirulina , Real-Time Polymerase Chain Reaction/methods , Newcastle Disease/diagnosis , Birds
Arq. bras. med. vet. zootec. (Online) ; 72(3): 1039-1046, May-June, 2020. ilus, tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1129747


O objetivo deste comunicado é desenvolver um método quantitativo PCR em tempo real, baseado em guia molecular (MB) (MB-qPCR) para detecção de infecção por espécies de Brucella, e avaliar seu potencial de utilização clínica. Os primers e as sondas MB foram desenhados para amplificação específica e determinação de sequência conservada do código do gene para os primeiros 58-aa da proteína de membrana externa OMP-2a, que é compartilhada em cinco espécies de Brucella epidêmicas. A avaliação metodológica foi realizada por análise de sensibilidade, especificidade, coeficiente de variação intra e inter, e a linearidade do qPCR. O potencial diagnóstico foi avaliado comparando-se o método qPCR desenvolvido com ensaios de exames bacteriológicos convencionais, incluindo os testes de soroaglutinação convencionais (SATs) e os testes do Rosa Bengala (RBPTs). O método exibiu alta sensibilidade (tão baixo quanto 50 cópias) e grande faixa de linearidade (102-108 cópias). Nenhuma reação cruzada foi encontrada com bactéria clínica comum. A sensibilidade diagnóstica foi superior ao exame bacteriológico, e a especificidade diagnóstica foi superior ao SAT ou ao RBPT. Um método MB-qPCR altamente sensível e específico para DNA de Brucella foi estabelecido com sucesso, provando ser uma ferramenta útil no diagnóstico molecular de brucelose.(AU)

Brucella/isolation & purification , Genome, Bacterial , Real-Time Polymerase Chain Reaction/methods
Gac. méd. Méx ; 156(3): 209-217, may.-jun. 2020. tab, graf
Article in English, Spanish | LILACS | ID: biblio-1249896


Resumen Introducción: A partir del 23 de marzo de 2020, en México se declaró la suspensión de actividades no esenciales en todo el país para mitigar la diseminación de la pandemia de COVID-19. Objetivo: Analizar los datos sobre los primeros 1510 casos de COVID-19 confirmados por laboratorio en México, describir la distribución geográfica de la enfermedad y su dinámica de transmisión. Método: Descripción de los primeros casos de COVID-19 con prueba positiva de RT-PCR en tiempo real, así como evaluación de las medidas epidemiológicas, incidencia acumulada, razón de contagios y tasas de mortalidad y letalidad durante el primer mes de la epidemia. Resultados: La edad promedio fue de 43 años y 58 % fue del sexo masculino; 44 % de los casos iniciales fue importado. La letalidad en la población durante el primer mes pasó de 1.08 a 3.97 por 100 casos; sin embargo, la tendencia es lineal y similar a la observada en Europa. Conclusiones: En México se está aplicando el distanciamiento social, pero aún se requieren estudios sobre la dinámica de la epidemia, la transmisión de persona a persona, la incidencia de infecciones subclínicas y la supervivencia de los enfermos.

Abstract Introduction As of March 23, 2020, suspension of non-essential activities was declared in Mexico throughout the country in order to mitigate the spread of the COVID-19 pandemic. Objective: To analyze data on the first 1510 laboratory-confirmed cases of COVID-19 in Mexico, and to describe the geographical distribution of the disease and its transmission dynamics. Method: Description of the first COVID-19 cases with real-time RT-PCR-positive test, as well as evaluation of epidemiological measures, cumulative incidence, rate of transmission, and mortality and lethality rates during the 1st month of the epidemic. Results: Average age was 43 years, and 58% were males; 44% of initial cases were imported. Lethality in the population during the 1st month went from 1.08 to 3.97 per 100 cases; however, the trend is linear and similar to that observed in Europe. Conclusions: In Mexico, social distancing is being applied, but studies are still required on the dynamics of the epidemic, person-to-person transmission, incidence of subclinical infections, and patient survival.

Humans , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Pneumonia, Viral/epidemiology , Social Isolation , Coronavirus Infections/epidemiology , Real-Time Polymerase Chain Reaction/methods , Pneumonia, Viral/transmission , Survival , Incidence , Coronavirus Infections/prevention & control , Coronavirus Infections/transmission , Pandemics/prevention & control , COVID-19 , Mexico/epidemiology
Rev. cuba. oftalmol ; 33(1): e775, ene.-mar. 2020. graf
Article in Spanish | LILACS, CUMED | ID: biblio-1126722


RESUMEN Objetivo: Estandarizar una técnica de reacción en cadena de la polimerasa en tiempo real para la detección del parásito e identificar Acanthamoeba en líquidos conservantes de lentes de contacto. Métodos: Se realizó un estudio descriptivo observacional de corte transversal sobre la técnica de reacción en cadena de la polimerasa en tiempo real para la detección de Acanthamoeba, en el Instituto de Investigaciones en Ciencias de la Salud de la ciudad de Asunción, en Paraguay. Se analizaron 110 líquidos conservantes aportados por usuarios sanos de lentes de contacto, mediante reacción en cadena de la polimerasa en tiempo real y cultivo en medio PAGE - SDS. Resultados: Se estandarizó con éxito la técnica de reacción en cadena de la polimerasa en tiempo real con límite de sensibilidad de 1 pg/µL. Se aisló Acanthamoeba a partir de una muestra (1 por ciento) por método de cultivo, mientras que la carga parasitaria en el líquido conservante fue inferior al límite de detección de la reacción en cadena de la polimerasa en tiempo real. El ADN obtenido del cultivo de dicha muestra fue positivo para Acanthamoeba por este método. Conclusión: El sistema estandarizado presenta buena sensibilidad y podrá ser incorporado en los laboratorios que cuentan con acceso a equipos de reacción en cadena de la polimerasa en tiempo real para un diagnóstico rápido y más eficiente en casos de sospechas de queratitis amebiana. Recomendamos el uso combinado de métodos moleculares y cultivo para aumentar la potencia del diagnóstico, sobre todo en muestras donde la carga parasitaria es muy baja(AU)

ABSTRACT Objective: Standardize a real-time polymerase chain reaction technique for detection of the parasite and identify Acanthamoeba in contact lens solutions. Methods: A cross-sectional observational descriptive study was conducted about a real-time polymerase chain reaction technique for detection of Acanthamoeba at the Institute of Health Sciences Research in the city of Asunción, Paraguay. A total 110 solutions were analyzed, which were provided by healthy contact lens users, by real-time polymerase chain reaction and culture in SDS-PAGE medium. Results: Successful standardization was achieved of the real-time polymerase chain reaction technique with a sensitivity limit of 1 pg/µl. Acanthamoeba was isolated from one sample (1 percent) by culture, whereas the parasite load in the contact lens solution was below the detection limit of the real-time polymerase chain reaction technique. The DNA obtained from the culture of that sample was positive for Acanthamoeba by the real-time polymerase chain reaction technique method. Conclusion: The system standardized exhibits good sensitivity and may be incorporated into laboratories with real-time polymerase chain reaction technique equipment for a rapid and more efficient diagnosis of suspected amoebic keratitis. We recommend the combined use of molecular methods and culture to enhance diagnostic power, mainly in samples where the parasite load is very low(AU)

Humans , Acanthamoeba/microbiology , Acanthamoeba Keratitis/etiology , Contact Lenses/adverse effects , Real-Time Polymerase Chain Reaction/methods , Epidemiology, Descriptive , Cross-Sectional Studies , Contact Lens Solutions/therapeutic use , Observational Studies as Topic
Rev. cuba. med ; 59(1): e1320, ene.-mar. 2020. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1139042


Introducción: La enfermedad producida por el Coronavirus 2019 ha impactado al mundo más allá de los límites de la salud pública a tres meses del diagnóstico del primer caso en China. En la actualidad afecta a 182 países, por lo que fue declarada pandemia por la Organización Mundial de la Salud. Objetivo: Revisar los aspectos clínico-epidemiológicos más importantes reportados sobre esta enfermedad. Desarrollo: La enfermedad se trasmite por las microgotas de saliva entre personas y por contacto con superficies contaminadas o heces. El periodo de incubación es hasta 14 días, en los cuales puede ocurrir trasmisión viral. Se caracteriza por un espectro sintomático variable, los pacientes pueden permanecer asintomáticos o presentar síntomas como fiebre, dolor de garganta, tos, anosmia y disnea. Aproximadamente 14 por ciento de los casos presenta formas graves y 5 por ciento evoluciona a estado crítico, asociado a complicaciones como el síndrome de distress respiratorio o shock séptico. El diagnóstico confirmatorio es a través de los estudios de reacción en cadena de polimerasa en tiempo real, en muestras respiratorias. No existe un tratamiento efectivo, aunque se emplean medicamentos, la mayoría de ellos como parte de ensayos clínicos. Conclusiones: Es un fenómeno sin precedentes en el último siglo en relación al número de personas y países que ha afectado y al impacto en las dinámicas sociales y económicas del mundo actual(AU)

Introduction: The disease caused by Coronavirus 2019 has impacted the world beyond the limits of public health three months after the first case was diagnosed in China. It currently affects 182 countries; hence it is was declared a pandemic by the World Health Organization. Objective: To review the most important clinical-epidemiological aspects reported on this disease. Findings: The disease is transmitted by droplets of saliva between people and by contact with contaminated surfaces or faeces. The incubation period is up to 14 days, in which viral transmission can occur. It is characterized by a variable symptom spectrum, patients can remain asymptomatic or present symptoms such as fever, sore throat, cough, anosmia and dyspnea. Approximately 14 percent of cases present severe forms and 5 percent progresses to a critical state, associated with complications such as respiratory distress syndrome or septic shock. The confirmatory diagnosis is through real-time polymerase chain reaction studies in respiratory samples. There is no effective treatment, although medications are used, most of them as part of clinical trials. Conclusions: It is, in the last century, an unprecedented phenomenon regarding the number of people and countries it has affected and concerning the impact on the social and economic dynamics currently in the world(AU)

Humans , Male , Female , Coronavirus Infections/diagnosis , Coronavirus Infections/prevention & control , Coronavirus Infections/epidemiology , Real-Time Polymerase Chain Reaction/methods , COVID-19/transmission , World Health Organization , eHealth Strategies
Braz. J. Pharm. Sci. (Online) ; 56: e18772, 2020. tab, graf
Article in English | LILACS | ID: biblio-1285509


There is emerging evidence for a dysregulation of insulin signaling in the brains of patients with Alzheimer's disease (AD) with overlapping molecular features to Type 2 Diabetes Mellitus (T2DM). In addition, T2DM is a known risk factor of AD. The goal of this study was to investigate the neurogenic and neuroprotective potential of rosmarinic acid (RA) in a streptozotocin (STZ)-induced combined with high fat diet (HFD) mouse model of diabetes. Animals were divided into four experimental groups (control, diabetic, diabetic + RA, RA only). Behavioral analysis was performed to assess spatial learning and anxiety levels of animals, whereas quantitative real time PCR was carried out to assess the gene expression levels of neuronal markers of neurogenesis (Ki67, DCX and NeuN). A significant decrease in memory and spatial learning was observed in the diabetic mice, which was substantially improved by RA treatment. RA also increased the gene expression of NeuN, DCX and Ki67, which were dysregulated in the diabetic model. This study proposes RA as a potential therapeutic agent to mitigate neuronal dysfunction associated with T2DM by promoting adult hippocampal neurogenesis.

Animals , Male , Mice , Diabetes Mellitus, Type 2/diagnosis , Alzheimer Disease/diagnosis , Risk Factors , Streptozocin/pharmacokinetics , Neurogenesis/genetics , Real-Time Polymerase Chain Reaction/methods
Article in English | LILACS, BBO | ID: biblio-1135545


Abstract Objective: To analyze whether the FokI polymorphism (rs228570) present in the vitamin D receptor gene in type 2 diabetics is related to chronic periodontitis's clinical status and evaluates the influence of chronic periodontitis on the perception of quality of life. Material and Methods: It is a clinical and laboratory study, composed of a sample of 59 individuals with previous diagnosis of type 2 Diabetes Mellitus and chronic periodontitis, of both sexes. On clinical examination, socio-epidemiological data and quality of life of patients with the Oral Health Impact Profile (OHIP-14) were recorded and a periogram was performed. Subsequently, saliva was collected spontaneously in sterile Falcon tubes (15 ml) and stored in the freezer at -20 °C. The purification of the genetic material was done with a PROMEGA kit (Wizard®), and the polymorphism studied was FokI (rs228570), found in the vitamin D receptor promoting region, with rs: 228570. After extraction of saliva DNA and purification, genotyping was performed by real-time PCR using specific allele probes (TaqMan® System). Results: The polymorphism of the vitamin D receptor gene was not positively associated with the severity and clinical characteristics of periodontitis, but suggested a relationship with the extent of the disease. Periodontitis also had no positive association with patients' perception of quality of life. Conclusion: The perception of quality of life of patients with chronic periodontitis and type 2 diabetes mellitus was compromised by the systemic condition, secondary to oral health, although some dimensions of OHIP-14 have been more frequently mentioned, such as psychological discomfort, physical pain and physical disability.

Humans , Polymorphism, Genetic , Quality of Life , Receptors, Calcitriol , Diabetes Mellitus, Type 2/diagnosis , Chronic Periodontitis/pathology , Periodontitis/pathology , Brazil , Chi-Square Distribution , Oral Health , Statistics, Nonparametric , Real-Time Polymerase Chain Reaction/methods
Article in English | LILACS, BBO | ID: biblio-1135530


Abstract Objective: To identify the occurrence of Veillonella spp. in children using real-time PCR (RT-PCR) and to determine its role as a risk factor for ECC in children aged 2-3 years. Material and Methods: A cross-sectional survey was conducted and samples from 87 children aged 2-3 years, who lived in selected villages in the Bandung City area, Indonesia, were collected. Examination for dental caries was performed using standard checks for decay, missing, and filled surfaces (dfms), and saliva samples were taken. Microbiological examination was performed using RT-PCR with primers consisting of one primary set for Veillonella spp. and one universal primary set for 16S rDNA. We performed statistical testing using the Mann Whitney rank-sum test. Results: A total of 87 children were sampled, and an ECC prevalence of 71.3% was found, with a mean dmfs of 7.1 (± 9.1). The proportion of Veillonella spp. in caries-free children was 2.13 ± 2.30, while in children with ECC, it was 3.29 ± 6.83. Conclusion: The proportion of Veillonella spp. in children with ECC was higher than in caries-free children; therefore, Veillonella spp. may be a risk factor for ECC.

Humans , Male , Female , Child, Preschool , Veillonella , Polymerase Chain Reaction/methods , Risk Factors , Dental Caries/prevention & control , Gram-Negative Bacteria/immunology , Epidemiologic Studies , Prevalence , Cross-Sectional Studies , Statistics, Nonparametric , Real-Time Polymerase Chain Reaction/methods , Indonesia/epidemiology
J. bras. nefrol ; 42(2,supl.1): 4-8, 2020. graf
Article in English | LILACS | ID: biblio-1134833


ABSTRACT The Covid-19 pandemic brought several challenges to the healthcare system: diagnosis, treatment and measures to prevent the spread of the disease. With the greater availability and variety of diagnostic tests, it is essential to properly interpret them. This paper intends to help dialysis units concerning the use of clinical criteria and diagnostic tests for decision making regarding the discontinuation of isolation of patients with suspected or confirmed Covid-19, as well as the return to work activities for employees with suspected or confirmed Covid-19.

RESUMO A pandemia da Covid-19 trouxe desafios ao sistema de saúde em diversas esferas: diagnóstico, tratamento e medidas para evitar a disseminação da doença. Com a maior disponibilização e variedades de testes diagnósticos, torna-se fundamental sua adequada interpretação. Este posicionamento pretende orientar unidades de diálise em relação ao uso de critérios clínicos e testes diagnósticos para a tomada de decisão referente à descontinuação do isolamento de pacientes com suspeita ou confirmação de Covid-19, assim como para o retorno às atividades laborais de colaboradores com suspeita ou confirmação de Covid-19.

Humans , Pneumonia, Viral/diagnosis , Renal Dialysis , Coronavirus Infections/diagnosis , Clinical Laboratory Techniques/standards , Return to Work , Betacoronavirus , Nephrology/standards , Patient Isolation , Pneumonia, Viral/epidemiology , Societies, Medical/standards , Algorithms , Brazil , Urology Department, Hospital/standards , Clinical Laboratory Techniques/methods , Checklist , Pandemics , Real-Time Polymerase Chain Reaction/methods , Clinical Decision-Making , COVID-19 Testing , SARS-CoV-2 , COVID-19
Rev. méd. Maule ; 34(2): 8-13, dic. 2019. tab
Article in English | LILACS | ID: biblio-1371186


INTRODUCTION: Increasing evidence suggests that changes in the balance of excitatory/inhibitory neurotransmission are involved in the development of the majority of chronic pain forms. In this context, impairment in glycine mediated inhibitory neurotransmission is thought to play a critical role in the disinhibition that accounts for the development and maintenance of central pain hypersensitivity. AIMS: The goal of this study was to evaluate the Glycine Receptor α3 subunit (α3GlyR) expression in neuropathic (Chronic Constriction Injury, CCI) and inflammatory (Zymosan A injected) animal models of chronic pain. RESULTS AND CONCLUSION: RT-qPCR analysis of spinal cord samples showed that glra3 gene expression does not change after 3 days of CCI and 4 hours of Zymosan A injection. However, we found that protein levels evaluated by Western blot increased after inflammatory pain. These data suggest that central sensitization is differentially regulated depending on the type of pain. α3GlyR protein expression plays an important role in the first step of inflammatory pain establishment.

Animals , Receptors, Glycine/metabolism , Receptors, Glycine/agonists , Central Nervous System Sensitization/physiology , Pain/diagnosis , Pain/physiopathology , Zymosan/administration & dosage , Pain Measurement/methods , Analysis of Variance , Receptors, Glycine/chemistry , Real-Time Polymerase Chain Reaction/methods
Rev. cuba. med. trop ; 71(1): e280, ene.-abr. 2019. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1093552


Se describe por primera vez una serie de nueve casos con clínica indicativa de leptospirosis en el municipio Puerto Nariño en el departamento Amazonas, Colombia. Se muestran evidencias serológicas de exposición con Rickettsia del grupo de las fiebres manchadas. Los casos fueron clínicamente considerados como síndrome febril de origen desconocido. Se descartó infección por dengue y malaria. El diagnóstico de Leptospira se realizó mediante el método de reacción en cadena de la polimerasa en tiempo real. Igualmente, se detectó la presencia de anticuerpos contra rickettsias del grupo de las fiebres manchadas por inmunofluorescencia Indirecta. Finalmente, se realiza revisión del tema(AU)

A description is provided for the first time of a series of nine cases with a clinical examination suggestive of leptospirosis in the municipality of Puerto Nariño, Department of Amazonas, Colombia. Serological evidence is presented of exposure to Rickettsia, spotted fever group. The cases were clinically considered as febrile syndrome of unknown origin. Infection with dengue or malaria was ruled out. Diagnosis of leptospirosis was achieved by real-time polymerase chain reaction. Additionally, indirect immunofluorescence detected the presence of antibodies against rickettsia, spotted fever group. Finally, a review was conducted about the topic(AU)

Humans , Adolescent , Adult , Middle Aged , Disease Outbreaks/prevention & control , Fluorescent Antibody Technique, Indirect/methods , Real-Time Polymerase Chain Reaction/methods , Leptospirosis/prevention & control , Leptospirosis/epidemiology , Fever/parasitology
Rev. Hosp. Clin. Univ. Chile ; 30(3): 207-217, 2019. tab, graf, ilus
Article in Spanish | LILACS | ID: biblio-1051370


V617F mutation in exon 14 of Janus Kinase 2 gene (jak-2) is used as a molecular marker for the diagnosis of Philadelphia negative myeloproliferative neoplasms (Phi-) such as Polycythemia Vera (PV), Essential Thrombocythemia (ET) and Primary Myelofibrosis (MFP). To detect this mutation, we used conventional polymerase chain reaction technique (PCR), a simple and inexpensive technique, however, has some drawbacks that current technology allows to solve. During the last years, more sensitive molecular techniques have been incorporated in clinical practice to support the diagnosis, prognosis and follow-up of hematological patients. For its implementation in the clinical routine should be considered technical and economic aspects, so in this work, we evaluate the Real Time PCR technique as a diagnostic method for the detection of the Jak-2-V617F mutation, using in house primers design. Our result show that the technique implemented has a concordance index of 0.87 with the conventional PCR used in the molecular diagnosis of myeloproliferative neoplasms. In addition, it has the same specificity, greater sensitivity and, shorter execution time in relation to conventional PCR. The implementation of this diagnostic method in our Hospital is technically possible and commercially convenient. (AU)

Humans , Janus Kinase 2/analysis , Real-Time Polymerase Chain Reaction/methods , Myeloproliferative Disorders/diagnosis , Real-Time Polymerase Chain Reaction/trends
Braz. j. microbiol ; 49(4): 909-913, Oct.-Dec. 2018. tab
Article in English | LILACS | ID: biblio-974301


ABSTRACT GeneXpert is one of the recent technological instruments used to diagnose tuberculosis in a short span of time. In this study, the performance of GeneXpert MTB/RIF assay for the diagnosis of extrapulmonary tuberculosis (EPTB) was compared with light-emitting diode Fluorescent Microscopy (LED-FM) in Khyber Pakhtunkhwa, Pakistan. A total of 737 EPTB samples were collected from tuberculosis (TB) suspected patients. Out of these samples, male to female ratio was 53% (n = 390) to 47% (n = 347) respectively. The sensitivity and specificity was 73% and 100% for GeneXpert, while 40% and 100% for LED-FM microscopy. This shows that the sensitivity of GeneXpert is 40-50%, higher than LED-FM microscopy. GeneXpert also detected low number of bacilli as compared to LED-FM microscopy.

Humans , Male , Female , Adult , Tuberculosis, Pulmonary/diagnosis , Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/methods , Microscopy/methods , Mycobacterium tuberculosis/isolation & purification , Pakistan , Tuberculosis, Pulmonary/microbiology , Middle Aged , Mycobacterium tuberculosis/classification , Mycobacterium tuberculosis/genetics