Comparing the multiplex RT-PCR method and liquichip technology in the detection of diarrhea-related virus

Abstract

Objective To compare the detection efficiency between multiplex RT-PCR method and liquichip technology for screening the viral etiological agents of diarrhea. Methods The development of the multiplex RT-PCR method. A total of 107 feces samples from patients who suffered from diarrhea and attended to Zhujiang Hospital of Southern University from September 2013 to February 2014 were collected and tested in parallel by both multiplex RT-PCR and xTAG Gastrointestinal Pathogen Panel (xTAG GPP) for Adenovirus, Norovirus genogroupⅠandⅡ, as well as by both multiplex RT-PCR and monoplex RT-PCR for Astrovirus and Sapovirus. To evaluate the sensitivity and specificity of multiplex RT-PCR, xTAG GPP and monoplex RT-PCR were used as reference.Kappa coefficient test was used to evaluate the consistency among the methods. The detection limit and accuracy of multiplex RT-PCR were evaluated by detection of serial dilution of positive plasmids and products sequencing for the five viral agents. Results The multiplex RT-PCR showed high consistency with xTAG GPP and monoplex RT-PCR, in which Kappa value was 0.885 and 1.000 respectively(P=0.000). Compared to xTAG GPP, the sensitivity and specificity of the multiplex RT-PCR were at average of 80.8%(21/26) and 100%(295/295)respectively. The detection limit and accuracy of multiplex RT-PCR were 104copies /μl-106copies/μl. Conclusion The high consistency indicated that both the multiplex RT-PCR and xTAG GPP are useful as a special, sensitive, high throughput and rapid diagnostic tools for the detection of the major viral pathogens related to diarrhea in clinical laboratory. (Chin J Lab Med, 2015, 38: 387-391) Key words: Diarrhea; Adenoviridae; Norovirus; Mamastrovirus; Multiplex polymerase chain reaction; Reverse transcriptase polymerase chain reaction