HAND system-based four multiplex RT-PCR for simultaneous detection of four diarrhea viruses / 南方医科大学学报
Journal of Southern Medical University
;
(12): 724-727, 2013.
Article
Dans Chinois
| WPRIM
| ID: wpr-306480
ABSTRACT
<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>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Virologie
/
ARN viral
/
Rotavirus
/
Astroviridae
/
RT-PCR
/
Norovirus
/
Sapovirus
/
Diarrhée
/
Fèces
/
Réaction de polymérisation en chaine multiplex
Type d'étude:
Etude diagnostique
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2013
Type:
Article
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