EV71 was Detected by Real-Time Fluorescence Quantitative RT-PCR Combined with Hand-Foot-Mouth Disease Rhesus Monkey Model

Abstract

In this study, a real-time fluorescence quantitative RT-PCR method was developed for the detection of enterovirus 71 (EV71) and the detection of EV71 viral load during the establishment of hand-foot-mouth disease (HFMD) model in rhesus monkeys. The model of hand-foot-mouth macaque was established by injecting newborn macaques infected with the virus venom, and the dynamic changes of virus content in blood samples from 0 to 14 days after infection were monitored by real-time quantitative RT-PCR. The linear range of the standard curve was between 103 and 108 copies/µL, and the PCR amplification rate was 99.37%. The results showed that the established method could detect a minimum of 102 copies of viral RNA, 100 times higher than the conventional RT-PCR method. It has the characteristics of strong specificity, high sensitivity and good stability. The results of blood sample analysis showed that the three groups of rhesus monkey models were successfully infected with EV71, and the copy number of virus proliferation was correlated with time and challenge dose. The results showed that real-time quantitative PCR could be used for quantitative detection of EV71 viral load.