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《Journal of Molecular Diagnostics and Therapy》 2018-05
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Establishment and clinical evaluation of a rapid real-time RT-PCR for detection of rotavirus A

CHEN Feng;TANG Xiaoyu;QIU Baofeng;LI Linzhong;YAN Hui;Nantong Entry-Exit Inspection and Quarantine Bureau;  
Objective To establish a rapid and early detection method of rotavirus A(RVA)by real-time reverse-transcription polymerase chain reaction(RT-PCR).Methods A pair of primers and a fluorescence probe based on conservative sequences of non-structural rotavirus protein 3(NSP3)in RVA genomes were designed to establish a RVA detection method by real-time RT-PCR,and then its sensitivity,specificity and precision were analyzed. A total of 200 cases from clinical stool samples were tested simultaneously and sequencing to evaluate the clinical performance of this method. Results The sensitivity of the method for detecting RVA reached 2.00 × 102 PFU/m.There was no cross-reactivity to other species viruses with similar species or similar symptoms.The coefficient of variation of Ct values of different concentrations of samples ranged from 0.82% to 2.28%. The results from 200 clinical samples showed that the total coincidence rate between the method and the sequencing method reached 98.5%. Conclusion The method established in this study has the characteristics of sensitive,specific,high precision,fast and simple,and has a good coincidence with sequencing methods.It has a good application prospect in the rapid diagnosis and epidemic monitoring of viral diarrhea.
【Fund】: 南通市科技计划项目(NO.MS12016030)
【CateGory Index】: R440
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