Development of a new method for diagnosis of Group B Coxsackie genome by reverse transcription loop-mediated isothermal amplification.
Indian J Med Microbiol
;
2011 Apr-June; 29(2): 110-117
Artículo
en Inglés
| IMSEAR
| ID: sea-143792
ABSTRACT
Background:
Coxsackie B viruses (genus, Enterovirus; family, Picornaviridae) can cause aseptic meningitis, encephalitis, pleurodynia, and fatal myocarditis, and are implicated in the pathogenesis of dilated cardiomyopathy. The differentiation of the group B Coxsackieviruses into their subtypes has potential clinical and epidemiological implications.Objective:
In this study, we developed a one-step, single-tube genogroup-specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of group B Coxsackie genomes targeting 5′ UTR region. Materials andMethods:
The amplification can be obtained in less than 1 hour by incubating all the reagents in a single tube with reverse transcriptase and Bst DNA polymerase at 63°C. Detection of gene amplification could be accomplished by agarose gel electrophoresis and the monitoring of gene amplification can also be visualised with the naked eye by using SYBR green I fluorescent dye.Results:
A total of 40 samples comprising 31 positive samples and 9 negative samples were used in this study for comparative evaluation. The results were compared with those from Real-Time Polymerase Chain Reaction (RT-PCR). None of the RT-PCR-positive samples were missed by RT-LAMP, thereby indicating a higher sensitivity of the RT-LAMP assay.Conclusion:
Thus, due to easy operation without a requirement of sophisticated equipment and skilled personnel, the RT-LAMP assay reported here is extremely rapid, cost-effective, highly sensitive, and specific and has potential usefulness for rapid detection of non-polio enterovirus (NPEV) not only by well-equipped laboratories but also by peripheral diagnostic laboratories with limited financial resources in developing countries.
Texto completo:
Disponible
Índice:
IMSEAR (Asia Sudoriental)
Asunto principal:
Compuestos Orgánicos
/
Coloración y Etiquetado
/
Temperatura
/
Factores de Tiempo
/
Humanos
/
ARN Viral
/
Sensibilidad y Especificidad
/
Enterovirus Humano B
/
Técnicas de Laboratorio Clínico
/
Infecciones por Coxsackievirus
Tipo de estudio:
Estudio diagnóstico
Idioma:
Inglés
Revista:
Indian J Med Microbiol
Asunto de la revista:
Microbiology
Año:
2011
Tipo del documento:
Artículo
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