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Indian Journal of Medical Microbiology
Medknow Publications on behalf of Indian Association of Medical Microbiology
ISSN: 0255-0857
EISSN: 0255-0857
Vol. 29, No. 2, 2011, pp. 110-117
Bioline Code: mb11028
Full paper language: English
Document type: Research Article
Document available free of charge

Indian Journal of Medical Microbiology, Vol. 29, No. 2, 2011, pp. 110-117

 en Development of a new method for diagnosis of Group B Coxsackie genome by reverse transcription loop-mediated isothermal amplification
Jaianand, K.; Saravanan, N.; Gunasekaran, P. & Sheriff, A. K.

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 and Methods: 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.

Keywords
Coxsackie B viruses, mediated isothermal amplification, non-polio enterovirus, real-time polymerase chain reaction, reverse transcription loop-mediated isothermal amplification

 
© Copyright 2011 Indian Journal of Medical Microbiology.
Alternative site location: http://www.ijmm.org

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