Abstract��Abstract: C4 is the mainly prevalent genotype of enterovirus 71 (EV71) in China��which can cause hand, foot and mouth disease. To develop a quantitative TaqMan PCR assay for the detection of EV71, a set of primers and TaqMan probe were designed based on the highly conserved region in VP1 gene of C4. The target gene was obtained and cloned into the vector pcDNA3.1+, and the transcribed RNA in vitro was diluted to get the standard RNA for setting up the standard curve. After the PCR conditions were optimized, the set of primers and probe were proved to be used for detecting the EV71 C4 subtype efficiently, and the optimal concentrations of the primers and probe were 300 nmol/L and 200 nmol/L, respectively. When viral load ranged from 1×103 to 1×109 copies, the R2 value was 1. The sensitivity of PCR reached 102 copies/μL. The inter- and intra-assay coefficient of variation was less than 0.5%, and no cross-reaction was found with coxsackievirus A16 (CA16), coxsackievirus B1 (CB1), human rotavirus (HRV) or herpes simplex virus type 2 (HSV-2). What′s more, six samples with EV71 were effectively identified to be positive. The results showed that the established quantitative PCR assay could provide a more rapid and useful way to diagnose and control hand, foot and mouth disease in China.
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������, �� ��, �� ��, �� ��, �Ű�÷, ��ѩɽ. ����Ҫ����EV71 VP1����߶ȱ�����Ϊ�е��TaqManӫ�ⶨ��PCR��ⷽ���Ľ���[J]. ������ѧ�о�, 2016, 20(3): 189-195.
HU Xiao-Xing, PENG Jie, FENG Yue, LIU Li, ZHANG A-Mei, XIA Xue-Shan. Development of a TaqMan Quantitative PCR Detection for the Predominant Enterovirus 71 Based on the Highly Conserved Region of VP1 Gene. Life Science Research, 2016, 20(3): 189-195.
2016, Vol. 20 
