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Journal of Southern Medical University ; (12): 724-727, 2013.
Artículo en Chino | WPRIM | ID: wpr-306480

RESUMEN

<p><b>OBJECTIVE</b>To establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.</p><p><b>METHODS</b>Primers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.</p><p><b>RESULTS</b>The 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.</p><p><b>CONCLUSION</b>The established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.</p>


Asunto(s)
Humanos , Astroviridae , Genética , Diarrea , Virología , Heces , Virología , Reacción en Cadena de la Polimerasa Multiplex , Métodos , Norovirus , Genética , ARN Viral , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Métodos , Rotavirus , Genética , Sapovirus , Genética
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