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2.
Rev. cuba. med. trop ; 73(2): e551, 2021. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1347496

ABSTRACT

El dengue es la infección trasmitida por vectores con mayor impacto en carga de enfermedad, económica y social a nivel mundial, con más de 3,6 billones de personas en riesgo de infección. Sus manifestaciones son variables, caracterizadas en su mayoría por síndrome febril con riesgo de sangrado, choque y muerte. El compromiso pulmonar es infrecuente, siendo el síndrome de dificultad respiratoria aguda una complicación inesperada, aunque informada, asociada a un mal pronóstico. Se presenta un paciente sin antecedentes relevantes de importancia, con focalización pulmonar severa asociado a infección por el virus dengue. En el caso presentado se descartaron procesos infecciosos bacterianos u otros agentes causales de síndrome de dificultad respiratoria aguda, lo que sumado a las características clínicas de ingreso, zona de ocurrencia del caso considerada como endémica, curso clínico, paraclínico y la franca positividad de las pruebas para dengue así como la seroconversión de estas, a pesar de no haber sido realizadas pruebas moleculares, se consideró como el agente causal más probable el virus dengue. Todo esto lleva a recomendar siempre considerarlo como potencial agente causal, lo que permite así un diagnóstico y manejo óptimos(AU)


Dengue is the vector-borne infection with the greatest impact on disease, economic and social burden worldwide, with more than 3.6 billion people under risk of contagion. Its manifestations are varied, most of them characterized by febrile syndrome with a risk of bleeding, shock and death. Pulmonary involvement is infrequent, and acute respiratory distress syndrome is an unexpected complication, though it has been reported in association to a bad prognosis. A case is presented of a male patient without relevant antecedents of interest, with severe pulmonary focalization associated to dengue virus infection. Bacterial infectious processes and other causative agents of acute respiratory distress syndrome were ruled out. In view of the patient's clinical status at admission, the endemicity of the patient's area of residence, the clinical and paraclinical course, and the obvious positivity of the dengue tests performed and their seroconversion, despite not having conducted molecular tests, it was concluded that the most probable causative agent was dengue virus. Therefore, it is recommended that dengue infection always be considered as a potential causative agent of acute respiratory distress syndrome, thus contributing to optimal diagnosis and management(AU)


Subject(s)
Humans , Male , Adult , Respiratory Distress Syndrome, Newborn , Endemic Diseases , Molecular Diagnostic Techniques , Dengue Virus , Prognosis , Cost of Illness , Seroconversion
3.
Med. infant ; 28(2): 105-109, Julio - Diciembre 2021. ilus
Article in Spanish | LILACS, BINACIS, UNISALUD | ID: biblio-1355205

ABSTRACT

Desde el inicio de la pandemia de COVID-19, el Laboratorio de Virología del Hospital Garrahan, implementó el diagnóstico molecular de SARS-CoV-2 mediante RT-PCR para dar respuesta rápida y de calidad a la creciente demanda. Al diagnóstico pediátrico se sumó el diagnóstico de los padres / acompañantes y personal de salud con criterio de caso sospechoso. Al inicio del 2021 se incorporó el test rápido de detección de antígeno para pacientes sintomáticos. Hasta junio de 2021 se procesó un total de 58 000 muestras para estudios moleculares. (AU)


Since the beginning of the COVID-19 pandemic, the Virology Laboratory of Garrahan Hospital has implemented molecular diagnosis of SARS-CoV-2 using RT-PCR in order to provide a rapid and high-quality response to the growing demand. In addition to the pediatric diagnosis, the diagnosis of parents/companions and healthcare personnel meeting the criteria of a suspected case was also added. At the beginning of 2021, the rapid antigen detection test for symptomatic patients was incorporated. Until June 2021, a total of 58,000 samples were analyzed by molecular studies. (AU)


Subject(s)
Humans , Laboratories, Hospital/statistics & numerical data , Molecular Diagnostic Techniques , COVID-19 Nucleic Acid Testing , COVID-19 Serological Testing , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , Pandemics
5.
Rev. bras. anal. clin ; 53(2): 175-179, 20210630. tab, ilus
Article in English | LILACS | ID: biblio-1353773

ABSTRACT

Objective: COVID-19 is presently the most serious public health concern and diagnosis is a principal tool for controlling and monitoring the spread of the disease. This study aimed to evaluate the efficiency of direct RT-PCR (dRT-PCR) for detection of SARS-CoV-2. Methods: Twenty-seven nasopharyngeal swabs from symptomatic individuals were evaluated. Standard RT-PCR was conducted, and for dRT-PCR the samples were preheated before amplification. Results: Positive agreement was 63.2% and negative agreement was 100%, being moderately in accord. Conclusion: dRT-PCR may be an alternative for screening symptomatic patients and a reliable option during an eventual shortage of viral RNA purification kits.


Objetivo: A COVID-19 é atualmente um sério problema de saúde pública e o diagnóstico é a principal ferramenta para controlar e monitorar a propagação da doença. Este estudo teve como objetivo avaliar a eficiência da RT-PCR direta (dRT-PCR) para detecção do SARS-CoV-2. Métodos: Vinte e sete amostras de swab nasofaríngeo de indivíduos sintomáticos foram avaliados. A RT-PCR padrão foi realizada e para a dRT-PCR as amostras foram pré-aquecidas antes da amplificação. Resultados: A concordância positiva foi de 63,2% e a concordância negativa foi de 100%, sendo moderadamente concordante. Conclusão: A dRT-PCR pode ser uma alternativa para a triagem de pacientes sintomáticos e uma opção confiável durante uma eventual escassez de kits de purificação de RNA viral.


Subject(s)
Humans , Virology , Polymerase Chain Reaction , Triage , Molecular Diagnostic Techniques , COVID-19 Nucleic Acid Testing , COVID-19
6.
Rev. méd. Urug ; 37(1): e37106, mar. 2021. tab
Article in Spanish | LILACS, BNUY | ID: biblio-1289843

ABSTRACT

Resumen: A nivel mundial se estima que en 2018 hubo alrededor de 10 millones de nuevos casos de tuberculosis (TBC). La detección molecular es una herramienta diagnóstica crecientemente utilizada para el diagnóstico de TBC. Los predictores de riesgo para TBC pulmonar son variados y varían de acuerdo a la población estudiada. Los objetivos del presente trabajo fueron: evaluar la performance de la detección de M. tuberculosis por la técnica Xpert® MTB/RIF para el diagnóstico de TBC pulmonar y determinar los factores predictores de presencia de esta enfermedad en pacientes asistidos en el Hospital Pasteur de Montevideo. Se realizó un estudio descriptivo, observacional y transversal. Se incluyeron 254 pacientes, 68 con TBC pulmonar. La sensibilidad de la prueba Xpert® MTB/RIF para detectar M. tuberculosis fue 100% (IC 95%: 91,2-100) y la especificidad 95,1% (IC 95%: 83,9-98,7). En el análisis multivariado se evidenció que los predictores independientes para presencia de TBC pulmonar fueron: contacto cercano con otro caso de TBC (p<0,001), consumo de pasta base de cocaína (p=0,006) y presentarse con adelgazamiento (p<0,001). En suma, la prueba Xpert® MTB/RIF se comportó como una excelente herramienta diagnóstica en nuestra población con elevada prevalencia de TBC pulmonar. Los predictores independientes para esta enfermedad indican que en la población analizada las estrategias de control de esta enfermedad requieren un abordaje multidisciplinario.


Summary: According to global estimations, there were approximately 10 million new cases of tuberculosis in 2018. Molecular diagnosis constitutes a rapidly growing diagnostic tool for tuberculosis. Risk predictors for pulmonary tuberculosis are varied and they depend on the population studied. The study aimed to assess the performance of M. tuberculosis detection by use of Xpert® MTB/RIF diagnostic test to diagnose pulmonary tuberculosis and to identify predictive factors for this disease in patients assisted at Pasteur Hospital in Montevideo. A descriptive, observational and transversal study was conducted, which included 254 patients, 68 of which had pulmonary tuberculosis. Sensitivity of the Xpert MTB/RIF assay to detect M. tuberculosis was 100% (CI 95%: 91.2-100) and specificity 95.1% (CI 95%: 83.9-98.7). Multivariate analysis evidenced the following to be the independent predictors that detect pulmonary tuberculosis: close contact with other cases of tuberculosis (p<0.001), coca-paste consumption (p=0.006) and evidence of loss of weight (p<0,001). To sum up, the Xpert® MTB/RIF assay proved to be an excellent diagnostic tool in our population with a high prevalence of pulmonary tuberculosis. Independent predictors for this disease show that, in the population studied, control strategies require a multidisciplinary approach.


Resumo: Globalmente, estima-se que em 2018 ocorreram cerca de 10 milhões de novos casos de tuberculose (TB). A detecção molecular é uma ferramenta diagnóstica cada vez mais usada para seu diagnóstico. Os preditores de risco para TB pulmonar são diversos e variam de acordo com a população estudada. Os objetivos deste estudo foram: avaliar o desempenho da detecção do M. tuberculosis pela técnica Xpert MTB/RIF para o diagnóstico da TB pulmonar e determinar os fatores preditivos da presença desta doença em pacientes atendidos no Hospital Pasteur de Montevidéu. Foi realizado um estudo descritivo, observacional e transversal. 254 pacientes foram incluídos, 68 com TB pulmonar. A sensibilidade do teste Xpert® MTB/RIF para detectar M. tuberculosis foi de 100% (IC 95%: 91,2-100) e a especificidade de 95,1% (IC 95%: 83,9- 98,7). A análise multivariada mostrou que os preditores independentes para a presença de tuberculose pulmonar foram: contato próximo com outro caso de tuberculose (p <0,001), consumo de pasta base de cocaína (p = 0,006) e apresentar perda de peso (p <0,001). Em suma, o teste Xpert® MTB/RIF se comportou como uma excelente ferramenta diagnóstica em nossa população com alta prevalência de TB pulmonar. Os preditores independentes para essa doença indicam que, na população analisada, as estratégias de controle da doença requerem uma abordagem multidisciplinar.


Subject(s)
Humans , Tuberculosis, Pulmonary/diagnosis , Molecular Diagnostic Techniques , Mycobacterium tuberculosis , Predictive Value of Tests , Sensitivity and Specificity
7.
Med. infant ; 28(1): 23-26, Marzo 2021. ilus, Tab
Article in Spanish | LILACS, BINACIS, UNISALUD | ID: biblio-1282888

ABSTRACT

Pneumocystis jirovecii es un hongo oportunista, causante de neumonía en huéspedes inmunocomprometidos. Es una infección grave con elevada tasa de mortalidad en pacientes oncohematológicos y receptores de trasplante de células progenitoras hematopoyéticas. La administración de corticosteroides es el principal factor de riesgo para adquirir esta infección. Actualmente las infecciones ocurren en aquellos pacientes que no reciben adecuada profilaxis. Las técnicas de diagnóstico molecular son las recomendadas por su elevada sensibilidad, especificidad y rapidez. La frecuencia global de P. jirovecii en pacientes inmunocomprometidos de nuestro hospital, durante el período evaluado fue de 4,8%, con una mortalidad global del 20%. Como factores de mal pronóstico se reportan la presencia de coinfecciones y la necesidad de asistencia respiratoria mecánica. Es importante la sospecha precoz en pacientes de riesgo, confirmada con un diagnóstico preciso mediante métodos moleculares para una intervención adecuada y oportuna (AU)


Pneumocystis jirovecii is an opportunistic fungus, causing pneumonia in immunocompromised hosts. It is a severe infection with a high mortality rate in oncology/hematology patients and hematopoietic stem cell transplant recipients. The administration of corticosteroids is the main risk factor for acquiring this infection. Currently infections occur in patients who do not receive adequate prophylaxis. Molecular diagnostic techniques are recommended because of their high sensitivity, specificity, and speed. In the study period, the overall incidence of P. jirovecii in immunocompromised patients at our hospital was 4.8%, with an overall mortality rate of 20%. Factors of a poor prognosis are the presence of coinfections and the need for mechanical respiratory assistance. Early suspicion in high-risk patients is important to confirm the diagnosis through molecular studies and start adequate and early treatment (AU)


Subject(s)
Humans , Infant , Child, Preschool , Child , Adolescent , Polymerase Chain Reaction/methods , Pneumocystis Infections/diagnosis , Pneumocystis Infections/epidemiology , Immunocompromised Host , Molecular Diagnostic Techniques/methods , Pneumocystis carinii/isolation & purification , Hospitals, Pediatric/statistics & numerical data , Cross-Sectional Studies , Retrospective Studies
8.
Pesqui. vet. bras ; 41: e06717, 2021. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1250488

ABSTRACT

The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.(AU)


O bugio ruivo (Alouatta guariba clamitans) é uma espécie de primata amplamente distribuída na América do Sul. As infecções por protozoários são comuns em primatas. Entretanto, estudos sobre protozoários em primatas no Brasil são escassos, portanto o objetivo deste estudo foi pesquisar DNA dos protozoários Apicomplexa Neospora caninum, Sarcocystisspp. e Toxoplasma gondii em tecidos de A. guariba clamitans. A extração de DNA foi realizada em amostras de tecido do coração, cérebro, fígado, baço, pulmão e intestino de seis A. guariba clamitans oriundos de Santa Maria, Região Central do Rio Grande do Sul, Brasil. Foi realizada PCR convencional utilizando primers geral do gene 18S rRNA para Apicomplexa e também primers específicos para amplificação de DNA de Neospora spp.e Toxoplasma gondii. Todos os animais foram positivos no PCR geral para Apicomplexa e no sequenciamento genético confirmou-se a presença de DNA de Sarcocystis nos tecidos de quatro animais pertencentes a pelo menos duas espécies (S. neurona e S. gigantea), e DNA de T. gondii foi detectado nos outros dois animais. Uma amostra positiva para T. gondii foi caracterizada genotipicamente como atípico pela técnica de polimorfismo do comprimento do fragmento de restrição. Não foi detectado DNA de N. caninum nas amostras testadas. A presença de DNA de protozoários apicomplexa nos tecidos dos seis animais testados neste estudo destaca a importância dos bugios ruivos como mantenedores desses patógenos na natureza.(AU)


Subject(s)
Animals , Toxoplasma/pathogenicity , Polymerase Chain Reaction , Apicomplexa/pathogenicity , Alouatta/microbiology , Genotyping Techniques/veterinary , Animals, Wild/microbiology , Protozoan Infections/diagnosis , DNA, Protozoan , Molecular Diagnostic Techniques , Infections
9.
Article in English | WPRIM | ID: wpr-887737

ABSTRACT

Objective@#To evaluate multidrug resistant loop-mediated isothermal amplification (MDR-LAMP) assay for the early diagnosis of multidrug-resistant tuberculosis and to compare the mutation patterns associated with the @*Methods@#MDR-LAMP assay was evaluated using 100 @*Results@#The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of MDR-LAMP were 85.5%, 93.6%, 96.7%, and 74.4% for the detection of resistance to isoniazid and rifampicin, respectively, and 80.5%, 92.3%, 98.6%, and 41.4% for the detection of @*Conclusion@#MDR-LAMP is a rapid and accessible assay for the laboratory identification of rifampicin and isoniazid resistance of


Subject(s)
Antitubercular Agents , Bacterial Proteins/genetics , Catalase/genetics , DNA, Bacterial/analysis , DNA-Directed RNA Polymerases/genetics , Drug Resistance, Multiple, Bacterial/genetics , Isoniazid , Molecular Diagnostic Techniques/methods , Mutation , Mycobacterium tuberculosis/isolation & purification , Nucleic Acid Amplification Techniques/methods , Oxidoreductases/genetics , Phenotype , Rifampin , Whole Genome Sequencing
10.
Arq. Inst. Biol ; 88: e00592020, 2021. ilus, tab
Article in English | LILACS, VETINDEX | ID: biblio-1357869

ABSTRACT

The diagnosis of bovine tuberculosis (TB) by molecular techniques has been broadly studied. These methods allow accelerating the diagnosis, in addition to presenting high specificity and sensitivity in the identification of the pathogen, critical characteristic for public health, especially when it comes to the direct diagnosis of the biologic samples, which has been little explored. This paper has evaluated a multiplex polymerase chain reaction (mPCR) as a tool to diagnose TB, which was performed directly on the granulomatous material of suspicious lesions collected in a cold chamber under state inspection in the state of Bahia, Brazil. Of the 74 samples evaluated, 14.86% were positive, with 10.81% positive for mPCR and culture, 4.05% negative for cultivation and positive for mPCR. The correlation between the cultivation and the mPCR presented agreeance higher than 61.54% of the cases. The results have indicated that the protocol proved itself effective, fast and very promising in the surveillance in slaughterhouses for the diagnosis of tuberculosis directly from the granuloma.(AU)


Subject(s)
Animals , Cattle , Tuberculosis, Bovine/diagnosis , Diagnosis , Multiplex Polymerase Chain Reaction , Mycobacterium , Abattoirs , Molecular Diagnostic Techniques , Granuloma , Noxae
11.
Ciencia Tecnología y Salud ; 8(2): 232-244, 2021. il 27 c
Article in Spanish | LILACS-Express | LILACS, LIGCSA, DIGIUSAC | ID: biblio-1353229

ABSTRACT

El carcinoma colorrectal (CCR) es de las primeras causas de mortalidad del mundo, presentando Guatemala una incidencia anual de 7.4/millón de habitantes. El síndrome de Lynch se caracteriza clínicamente por un inicio temprano del CCR con lesiones causadas por alteraciones en genes que codifican proteínas reparadoras.Los microsatélites son regiones del ADN con una unidad repetitiva de uno o más nucleótidos y son susceptibles a errores durante la replicación de ADN de los enterocitos. Existe un sistema de reparación que corrige estos errores. Cuando las proteínas reparadoras de este sistema están mutadas o ausentes, dichos errores del ADN persisten. Estas proteínas reparadoras se expresan en el núcleo de las células colónicas normales y son detecta-bles utilizando estudios de inmunohistoquímica (IHQ). Los genes MLH1 y MSH2 pueden encontrarse mutados en el 90% de los casos de cáncer colorrectal y el resto corresponde a MSH6 y PMS2. Esta vía oncogénica se caracteriza por alteración del sistema de reparación de errores durante la replicación del ADN, controlado por los genes MMR (mismatch repair), principalmente MLH1, MSH2, MSH6 y PMS2. Se realizó una revisión extensa de la literatura en PubMed, Springer y JAMA, usando las palabras clave: fenotipo de CCR, Síndrome de Lynch e inestabilidad microsatelital, detectándose 55 artículos. El objetivo de esta revisión es describir la importancia de la identificación del fenotipo del CCR por medios de IHQ y de pruebas moleculares para el eficaz tratamiento con inmunoterapia anti-PD1/PD-L1.


Colorectal cancer (CRC) is one of the leading causes of mortality in the world. In Guatemala it's an important cause of morbidity (7.4 per million inhabitants). Lynch syndrome is clinically characterized by an early onset of nonpolyposis colorectal carcinoma, with multiple lesions and neoplasms. The syndrome is caused by mutations in genes encoding DNA mismatch repair proteins. The microsatellites are regions of the DNA that repeat between one or more nucleotides and are susceptible to errors during replication, these are corrected by a repair system, when genes are mutated, the errors persist. The genes encoding repair proteins are expressed in the nuclei of normal colonic cells which can be observed using immunohistochemical studies. The MLH1, MSH2 genes are found to be mutated in 90% of the cases and the rest corresponds to the MSH6 and PMS2 genes. This oncogenic pathway characteristically consists of an alteration in the DNA repair system that is controlled by mismatch repair genes (MMR). An extensive research was conducted on PubMed, Springer and JAMA, using the keyword: CRC phenotype, Lynch syndrome and microsatellite instability. 55 articles were found. This review«s objective is to understand the mechanisms of nonpolyposis colorectal cancer and the importance of identifying patients with a mutant phenotype as a predictive factor for the efficacy of the anti-PD1/PDL1 immunotherapy and for prognosis.


Subject(s)
Humans , Carcinoma/mortality , Colorectal Neoplasms/mortality , Microsatellite Instability , Immunohistochemistry , Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , Microsatellite Repeats , Enterocytes , Molecular Diagnostic Techniques , Mismatch Repair Endonuclease PMS2/genetics , MutL Protein Homolog 1/genetics , Mutation
12.
Med. lab ; 25(3): 565-567, 2021.
Article in Spanish | LILACS | ID: biblio-1343143

ABSTRACT

Los nódulos tiroideos siempre han sido considerados problemas comunes en la práctica clínica, y con el fin de estudiarlos se han buscado diferentes técnicas de aproximación y diagnóstico a lo largo de los años [1,2]. Esto ha contribuido al aumento en su incidencia, ya que a mayor número de estudios de imágenes realizados, aumenta la probabilidad de encontrarlos, puesto que tan solo el 50% son detectados por palpación al examen físico [3]. Es importante mencionar que a pesar de que solo el 5% de los nódulos tiroideos finalmente presentan un diagnóstico maligno y requieren manejo quirúrgico, hace algunos años se llevaban a cabo exámenes médicos diagnósticos y manejos quirúrgicos invasivos y exhaustivos que no eran prácticos, necesarios ni costo efectivos [4]. Fue solo hasta hace aproximadamente 40 años que se empezó a difundir el uso de la técnica diagnóstica desarrollada en 1950 por Söderstrom, Einhorn, Franzèn y Zajicek en el Hospital Radiumhelmet, de Estocolmo, Suecia, y considerada actualmente como la prueba de elección para la evaluación de nódulos tiroideos: la aspiración con aguja fina (ACAF), la cual se describe segura, precisa y rentable


Subject(s)
Humans , Thyroid Nodule , Cell Biology , Molecular Diagnostic Techniques
13.
Braz. arch. biol. technol ; 64: e21190423, 2021. tab, graf
Article in English | LILACS | ID: biblio-1285548

ABSTRACT

Abstract High sensitivity of qPCR assay can be compromised by the presence of PCR inhibitors in samples analyzed. The aim of this study was to analyze the RT-qPCR assay efficiency considering the RNA quality/quantity and the presence of PCR inhibitors in patients with chemotherapy and/or antibiotic therapy. We analyzed 60 samples using RT-qPCR from individuals suspected of leukemia and 44 samples were quantified by fluorimetry and spectrophotometry. The efficiency of the RT-qPCR assay was evaluated comparing the threshold cycle (Ct) from tested samples and the standard curve. The 260/280 and 260/230 ratios, the presence of PCR inhibitors and the amount of sample (ng) used in the RT-qPCR reaction can be associated with 56.8% (R²=0.56, p<0.05) in the Ct obtained. The decrease of the RT-qPCR efficiency can be explained in 42,8% due to the variation of the 260/280 ratio (R²=0.42,p<0.05). The presence of antibiotics in the blood sample can be associated in 11.3% with the variability of 260/280 ratio (R²=0.11,p<0.05). Presence of chemotherapeutic drugs in the blood sample was not correlated with Ct variation (p=0.17). The spectrophotometer determines a RNA quantification with 2.2 times higher than the fluorimeter (t=2.2, p=0,03) and this difference is correlated with the 260/280 ratio (R²=0.36, p<0.05). Samples with low purity had a reduction in the qPCR efficiency, although we did not observe false results.


Subject(s)
Humans , Polymerase Chain Reaction/methods , Nucleic Acid Synthesis Inhibitors , Molecular Diagnostic Techniques/methods , Spectrophotometry/instrumentation , Fluorometry/instrumentation
14.
Rev. bras. parasitol. vet ; 30(1): e020220, 2021. tab, graf
Article in English | LILACS | ID: biblio-1251358

ABSTRACT

Abstract Trypanosoma vivax infections cause nonspecific clinical signs in cattle associated with aparasitemic intervals, making disease diagnosis a challenge. In Brazil, diminazene aceturate and isometamidium chloride (ISM) are available to treat bovine trypanosomosis. The objective of this study was to follow-up, by molecular and serological techniques, dairy cattle naturally infected by T. vivax after ISM treatment. Thirty cattle naturally infected with T. vivax received two applications of ISM, at a dosage of 1.0 mg/kg intramuscularly, on days 0 and 150. For T. vivax diagnosis, EDTA-blood and serum samples were evaluated on 0, 7, 15, 30, 60, 90, 120, 150, 180, 210, and 240 days after treatment PCR, Loop-mediated isothermal amplification (LAMP) and ELISA. Animals with persistent detection of T. vivax DNA by both PCR and LAMP were found and continuous detection of anti-T. vivax IgG antibodies by ELISA, suggesting the presence of T. vivax resistance to ISM. The combination of LAMP and ELISA tests can prevent misdiagnosis of the parasite clearance in treated cattle, contributing to better disease control. This is the first experiment that demonstrates the persistence infection of T. vivax under ISM treatment in a natural infected herd and evidence of ISM chemotherapy-resistant T. vivax in Brazil.


Resumo Em bovinos, infecções por Trypanosoma vivax geram sinais clínicos inespecíficos que, associados a intervalos aparasitêmicos, faz com que o diagnóstico da enfermidade seja desafiador. No Brasil, somente aceturato de diaminazeno e cloridrato de isometamidum (ISM) estão disponíveis para o tratamento da tripanossomose bovina. Este trabalho teve como objetivo acompanhar bovinos leiteiros naturalmente infectados por T. vivax, após o tratamento com ISM por meio de técnicas moleculares e sorológica. Foram utilizados 30 bovinos naturalmente infectados com T. vivax, sendo estes tratados com duas aplicações de ISM, na dosagem de 1,0 mg/kg por via intramuscular profunda, nos dias 0 e 150. Foram avaliadas, para diagnóstico de T. vivax, amostras de sangue acrescido de EDTA e soro, colhidas nos 0, 7, 15, 30, 60, 90, 120, 150, 180, 210 e 240 dias após os tratamentos pela reação em cadeia da polimerase (PCR), amplificação circular isotérmica do DNA (LAMP) e ensaio de imunoabsorção enzimático (ELISA). Verificou-se a presença de animais com persistência na detecção de DNA de T. vivax pela PCR e LAMP, bem como detecção contínua de anticorpos IgG anti-T. vivax pelo método de ELISA, sugerindo a presença de resistência de T. vivax ao ISM. A combinação dos testes LAMP e ELISA pode evitar falsos diagnósticos da eliminação do parasita nos bovinos tratados, contribuindo para um melhor controle da doença. Este é o primeiro experimento que demonstra infecção persistente do T. vivax em rebanho naturalmente infectado, tratado com ISM, e evidencia possível resistência ao quimioterápico no Brasil.


Subject(s)
Animals , Trypanocidal Agents/therapeutic use , Trypanosomiasis, African/veterinary , Trypanosomiasis, Bovine/diagnosis , Trypanosomiasis, Bovine/drug therapy , Phenanthridines , Brazil , Cattle , Follow-Up Studies , Trypanosoma vivax , Nucleic Acid Amplification Techniques , Molecular Diagnostic Techniques
15.
Malar. j. (Online) ; 20(390): 1-12, 2021. Mapas, Tab.
Article in English | AIM, RSDM, AIM | ID: biblio-1352541

ABSTRACT

Background: Artemisinin-based combination therapy (ACT) has been the recommended first-line treatment for uncomplicated malaria in Mozambique since 2006, with artemether­lumefantrine (AL) and amodiaquine­artesunate (AS­AQ) as the first choice. To assess efficacy of currently used ACT, an in vivo therapeutic efficacy study was conducted. Methods: The study was conducted in four sentinel sites: Montepuez, Moatize, Mopeia and Massinga. Patients between 6 and 59 months old with uncomplicated Plasmodium falciparum malaria (2000­200,000 parasites/µl) were enrolled between February and September of 2018, assigned to either an AL or AS­AQ treatment arm, and monitored for 28 days. A Bayesian algorithm was applied to differentiate recrudescence from new infection using genotyping data of seven neutral microsatellites. Uncorrected and PCR-corrected efficacy results at day 28 were calculated. Results: Totals of 368 and 273 patients were enrolled in the AL and AS­AQ arms, respectively. Of these, 9.5% (35/368) and 5.1% (14/273) were lost to follow-up in the AL and AS­AQ arms, respectively. There were 48 and 3 recurrent malaria infections (late clinical and late parasitological failures) in the AL and AS­AQ arms, respectively. The day 28 uncorrected efficacy was 85.6% (95% confidence interval (CI) 81.3­89.2%) for AL and 98.8% (95% CI 96.7­99.8%) for AS­AQ, whereas day 28 PCR-corrected efficacy was 97.9% (95% CI 95.6­99.2%) for AL and 99.6% (95% CI 97.9­100%) for AS­AQ. Molecular testing confirmed that 87.4% (42/48) and 33.3% (1/3) of participants with a recurrent malaria infection in the AL and AS­AQ arms were new infections; an expected finding in a high malaria transmission area. Adverse events were documented in less than 2% of participants for both drugs. Conclusion: Both AL and AS­AQ have therapeutic efficacies well above the 90% WHO recommended threshold and remain well-tolerated in Mozambique. Routine monitoring of therapeutic efficacy should continue to ensure the treatments remain efficacious.


Subject(s)
Child, Preschool , Malaria, Falciparum , Malaria/drug therapy , Parasites , Patients , Recurrence , Safety , Therapeutics , Algorithms , Polymerase Chain Reaction , Efficacy/methods , Monitoring , Molecular Diagnostic Techniques , Lost to Follow-Up , Artesunate/administration & dosage , Artemether/administration & dosage , Lumefantrine , Infections , Mozambique/epidemiology
16.
Rev. med. Risaralda ; 26(2): 160-165, jul.-dic. 2020. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1150025

ABSTRACT

Resumen El síndrome de Ellis van Creveld es un trastorno autosómico recesivo, caracterizado por mutaciones en los genes ECV y ECV2, los cuales son importantes para el desarrollo osteocondral. A nivel mundial, se han reportado aproximadamente 300 casos ,presentándose con mayor frecuencia en poblaciones endogámicas. Se caracteriza por distrofias óseas, displasias ectodérmicas y malformaciones cardíacas. El diagnóstico clínico puede ser confirmado mediante pruebas moleculares. A continuación, se presenta el caso de una paciente diagnosticada con el síndrome, la cual fue evaluada de manera interdisciplinaria. Esta revisión permitió dar a conocer un nuevo caso de la patología, relacionar las manifestaciones clínicas de la paciente con la literatura y describir nuevos hallazgos que pueden correlacionarse con el síndrome.


Abstract Ellis Van Creveld syndrome is an autosomal recessive disorder, characterized by mutations of the genes ECV and ECV2, which are very important in the osteochondral development. Worldwide, there have been reported around 300 cases that are commonly evidenced in populations where endogamy is typical. It is clinically characterized by bone dystrophies, ectodermal dysplasias, and congenital heart defects; the diagnosis can be confirmed by molecular tests. In the lines below, a case of a patient that suffers from this syndrome, and that was examined in an interdisciplinary way will be presented. This review allows us to show a new case of this pathology, to relate the clinical symptoms of the patient with the existing literature, and to describe new findings that can be correlated with the Ellis Van Creveld condition.


Subject(s)
Humans , Female , Child , Congenital Abnormalities , Ellis-Van Creveld Syndrome , Signs and Symptoms , Volition , Ectodermal Dysplasia , Molecular Diagnostic Techniques , Genes , Heart Defects, Congenital , Mutation
17.
Infectio ; 24(4): 224-228, oct.-dic. 2020. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1114873

ABSTRACT

Resumen Objetivo: Comparar los resultados obtenidos de diferentes sistemas de identificación de C. auris. Métodos: Análisis descriptivo con datos recopilados durante 2016-19 mediante la vigilancia nacional. Se evaluaron los resultados generados por los sistemas MicroScan, Phoenix BD, VITEK 2 y MALDI-TOF MS de instituciones hospitalarias de 843 aislamientos clínicos sospechosos de C. auris remitidos al INS y se compararon con los resultados generados de confirmación a través de MALDI- TOF MS (Bruker Daltonics) o PCR. Resultados: De los 843 aislamientos clínicos remitidos al INS, el 81,7% fueron confirmados como C. auris mediante MALDI- TOF MS o PCR en el INS y el resto, 18,3%, fueron identificados como otras especies de Candida spp. Las identificaciones correctas enviadas por los laboratorios representaron el 42,4%. MicroScan identificó C. auris principalmente como C. haemulonii, C. guilliermondii, C. albicans y C. famata; Phoenix BD, VITEK 2 y MALDI-TOF MS identificó C. auris como C. haemulonii. Discusión: Estudios señalan que C. auris exhibe una estrecha relación filogenética con C. haemulonii. Las identificaciones discrepantes pueden darse debido a que las bases de datos de los sistemas de diagnóstico son limitadas para este patógeno. Las deficiencias de los sistemas comerciales para la identificación de C. auris deben ser complementados con otros sistemas como MALDI-TOF MS o pruebas moleculares.


Abstract Objective: To compare the identification results obtained by different identification systems of C. auris isolates. Methods: A descriptive study with data collected during the years 2016-19 through surveillance. The results generated by the MicroScan, Phoenix BD, VITEK 2 and MALDI-TOF MS systems of 843 clinical isolates of C. auris submitted to the INS were evaluated and compared with the results generated from confirmation through MALDI-TOF MS (Bruker Daltonics) or PCR. Results: Out of 843 clinical isolates submitted to the INS, 81.7% were confirmed as C. auris by MALDITOF MS or PCR in the INS and the rest, 18.3%, were identified as other species of Candida spp. The correct identifications sent by the laboratories was 42.4%. MicroScan identified C. auris as C. haemulonii, C. guilliermondii, C. albicans and C. famata; Phoenix BD, VITEK 2 and MALDI-TOF MS identified C. auris as C. haemulonii. Discussion: Studies indicate that C. auris exhibits a close phylogenetic relationship with C. haemulonii. In addition, discrepant identifications may occur because the databases of diagnostic systems are limited with reference to this pathogen. The deficiencies of commercial systems for the identification of C. auris must be complemented with other systems such as MALDI-TOF MS or molecular tests.


Subject(s)
Humans , Female , Candida , Surveillance , Diagnosis , Laboratories , Polymerase Chain Reaction , Epidemiology, Descriptive , Colombia , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Molecular Diagnostic Techniques , Alkalies
18.
Infectio ; 24(4): 208-211, oct.-dic. 2020. graf
Article in English | LILACS, COLNAL | ID: biblio-1114870

ABSTRACT

Abstract Objective: The aim of the study was detection of two major causative agents of pleuropneumonia, Mycoplasma capricolum subsp. capripneumoniae (Mccp) and Mannheimia haemolytica, in goats. To the best of our knowledge, this study is the first investigation of Mccp in Iran. Methods: 50 grossly suspected lungs to pleuropneumonia and 10 healthy samples were collected from Shiraz abattoir. Results: Histopathological evaluation of tissue samples showed various diagnosed pneumonias including 40% bronchointerstitial pneumonia (20 samples), 34% interstitial pneumonia (17 samples), 10% fibrinopurulent bronchopneumonia (5 samples), 12% purulent bronchopneumonia (6 samples) and 4% chronic pneumonia (2 samples). In molecular study, all 50 suspected samples and 10 healthy ones by PCR showed no Mccp positive sample, but the detection rate of M. haemolytica in suspected samples was 14% and in healthy lungs was zero. Conclusions: It may be concluded that goats referred to Shiraz abattoir is free of Mccp. Further sampling and molecular testing at the level of suspected herds to CCPP can be useful.


Resumen Objetivo: El objetivo del estudio fue la detección de dos agentes causantes principales de pleuroneumonía, Mycoplasma capricolum subsp. Capripneumoniae (Mccp) y Mannheimia haemolytica, en cabras. Hasta donde sabemos, este estudio es la primera investigación de Mccp en Irán. Métodos: 50 pulmones muy sospechosos de pleuroneumonía y 10 muestras sanas se obtuvieron del matadero de Shiraz. Resultados: La evaluación histopatológica de muestras de tejido mostró varias neumonías diagnosticadas, incluyendo 40% de neumonía broncointersticial (20 muestras), 34% de neumonía intersticial (17 muestras), 10% de bronconeumonía fibrinopurulenta (5 muestras), 12% de bronconeumonía purulenta (6 muestras) y 4% neumonía crónica (2 muestras). En un estudio molecular, las 50 muestras sospechosas y 10 sanas por PCR no mostraron una muestra positiva de Mccp, pero la tasa de detección de M. haemolytica en muestras sospechosas fue del 14% y en pulmones sanos fue cero. Conclusiones: se puede concluir que las cabras referidas al matadero Shiraz están libres de Mccp. La realización de muestreo adicional y pruebas moleculares a nivel de rebaños sospechosos para CCPP puede ser útil.


Subject(s)
Animals , Pleuropneumonia , Goats , Mannheimia haemolytica , Mycoplasma capricolum , Pneumonia , Bronchopneumonia , Abattoirs , Lung Diseases, Interstitial , Molecular Diagnostic Techniques , Methods
19.
Rev. cuba. med. trop ; 72(3): e532, sept.-dic. 2020. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1156544

ABSTRACT

Introducción: En la actualidad las infecciones fúngicas representan un problema para la salud humana. Las infecciones causadas por especies patógenas de hongos registran un incremento constante y se ubican entre el cuarto y décimo lugar como causa de muerte, particularmente en las unidades de cuidado intensivo. Un diagnóstico adecuado y precoz impacta directamente en la morbilidad y mortalidad asociadas a estas. Objetivo: Describir las principales técnicas de diagnóstico no convencional de las enfermedades fúngicas más frecuentes, en especial las relacionadas con el diagnóstico serológico y molecular. Métodos: Se realizó una revisión de la literatura científica sobre el tema, publicada entre 2000 y 2019. Se revisaron un total de 63 trabajos. Como motores de búsqueda se emplearon Google y Google Scholar. Se revisaron las bases de datos Medline, PubMed, Science Direct, BUCea y SciELO. Análisis y síntesis de la información: Las técnicas serológicas se emplean en el diagnóstico de las micosis invasivas o sistémicas por ser fáciles, rápidas y confiables. La detección de anticuerpos tiene utilidad limitada en el diagnóstico de las micosis invasivas debido a que la respuesta puede estar retrasada, reducida o no existir en pacientes inmunocomprometidos. La detección de componentes no antigénicos liberados por los hongos durante la infección y la secuenciación de ácidos nucleicos fúngicos son otras opciones para el diagnóstico de las micosis. Conclusiones: El desarrollo biotecnológico aporta nuevas herramientas que incrementan las oportunidades de identificación de las micosis. En la actualidad se disponen de métodos basados tanto en la detección de marcadores inmunológicos como de elementos moleculares específicos. La eficacia de las herramientas no convencionales para el diagnóstico depende de la correcta combinación de estas(AU)


Introduction: Fungal infections are a current human health problem. Infections caused by pathogenic fungal species constantly increase in number, and are ranked between the fourth and tenth leading causes of death, particularly in intensive care units. Early accurate diagnosis has a direct impact on the morbidity and mortality of fungal infections. Objective: Describe the main non-conventional diagnostic techniques for the most common fungal diseases, especially those related to serological and molecular diagnosis. Methods: A review was conducted of the scientific literature about the topic published between the years 2000 and 2019. A total 63 publications were reviewed. The search engines used were Google and Google Scholar. The databases Medline, PubMed, Science Direct, BUCea and SciELO were reviewed. Data analysis and synthesis: Serological techniques are used for the diagnosis of invasive or systemic mycoses because they are easy, fast and reliable. The detection of antibodies has a limited usefulness in invasive mycosis diagnosis, for the response may be delayed, reduced or inexistent in immunocompromised patients. Detection of non-antigenic components released by fungi during infection and sequencing of fungal nucleic acids are other mycosis diagnosis options. Conclusions: Biotechnological development contributes new tools increasing mycosis identification opportunities. Methods are currently available which are based on detection of immunological markers and specific molecular elements. The efficacy of non-conventional diagnostic tools depends on their appropriate combination(AU)


Subject(s)
Humans , Molecular Diagnostic Techniques/methods , Mycoses/diagnosis , Mycoses/mortality , Clinical Laboratory Techniques/methods
20.
Biomédica (Bogotá) ; 40(4): 626-640, oct.-dic. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1142429

ABSTRACT

Resumen. Introducción. La prueba Xpert MTB/RIF™ es una prueba molecular rápida para el diagnóstico de la tuberculosis y la resistencia a la rifampicina. Desde el 2010 es la recomendada por la Organización Mundial de la Salud (OMS) y, aunque fue introducida en Colombia en el 2012, se desconocen los resultados de su uso.Objetivo. Describir la cobertura y la fidelidad en el uso de la prueba Xpert MTB/RIF™ en pacientes con tuberculosis pulmonar en una ciudad con alta carga de la enfermedad en Colombia.Materiales y métodos. Se hizo un estudio retrospectivo descriptivo de casos del programa de tuberculosis en Cali entre el 2013 y el 2019. La cobertura se estimó como el total de pruebas empleadas en los casos registrados en el programa. La fidelidad se midió con base en los protocolos internacionales de uso de la Xpert MTB/RIF™. Además, se hizo un análisis de correspondencias múltiples entre la prueba y las variables sociodemográficas.Resultados. Se incluyeron 6.328 pacientes con tuberculosis pulmonar, de los cuales 181 eran resistentes a los fármacos. La cobertura total de la Xpert MTB/RIF™ durante el periodo de estudio fue de 10,3 % (n=655), con una variación anual entre 0,2 y 23 %. La fidelidad fue de 46,8 % para los grupos de mayor riesgo de tuberculosis multirresistente (TB-MDR). El uso de la prueba se relacionó con la condición de ser hombre, afrocolombiano, y tener entre 41 y 60 años de edad.Conclusiones. La cobertura de la prueba Xpert MTB/RIF™ en Cali es baja y su uso no responde a la priorización recomendada para su implementación. Se requieren estrategias para promover su uso adecuado, de manera que contribuya a la meta de poner fin a la tuberculosis.


Abstract. Introduction:The Xpert MTB/RIF™ is a rapid molecular test that diagnoses tuberculosis and rifampin resistance. Since 2010, it is recommended by the World Health Organization (WHO) and although it was introduced in Colombia since 2012, the results of its implementation are unknown.Objective: To describe the coverage and fidelity in the implementation of the Xpert MTB/RIF™ in patients with pulmonary tuberculosis in a city with a high burden for the disease in Colombia.Materials and methods: We conducted a retrospective, descriptive study of cases from a tuberculosis program in Cali between 2013 and 2019. We estimated the coverage as the total number of tests used compared to the cases registered in the program and the fidelity based on international Xpert MTB/RIF™ implementation protocols. We performed a multivariate analysis of multiple correspondences between the test and the sociodemographic variables.Results: We included 6,328 patients with pulmonary tuberculosis of whom 181 were drug-resistant. The Xpert MTB/RIF™ coverage was 10,3% (n=655) with an annual variation between 0.2% and 23%. Loyalty among the highest risk groups of MDR-TB was 46.8%. The use of the test was related to being an Afro-Colombian man between 41 and 60 years of age.Conclusions: The coverage of the Xpert MTB/RIF in Cali is low and its use does not follow the recommended prioritization for its implementation. Implementation strategies are required for its proper use to contribute to the goal of ending tuberculosis.


Subject(s)
Tuberculosis, Pulmonary , Molecular Diagnostic Techniques , Rifampin , Drug Resistance
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