Fluorescent quantitative RT-PCR detection reagent box and detection method of enterovirus EV71

Abstract

The invention provides an intestinal tract virus EV71 fluorescence RT-PCR detection kit and a detection method applied to the emergency detection in a hand-foot-and-mouth disease laboratory. The primer and the probe sequences of the fluorescent quantitation RT-PCR detection kit are: EV 71 YG F1:5'-TGA TTGAGACACGC / GTGTGTT / CCTTA-3'; EV71 YG R1:5'-CCCGC TCTGCTGAAGAAACT-3'; and EV71YG pb 1:5'-TCGCACAGCACA GCTGAGACCACTC-3'. The method has high degree of specificity on the detection of the intestinal tract virus EV71 and does not have cross reaction with virus such as CA16, CA4, CA21, E6, E30, CoxB5 and POLIO. The detection sensitivity of the method is up to 0.1TCID50. The method can detect the intestinal tract virus EV71 nucleic acid directly from the samples such as the ncurolymph, the herpes fluid and the night soil of the suspect patient with the hand-foot-and-mouth disease. The process from extracting the nucleic acid from the virus to completing the detection only needs three hours.

Description

(1) Technical field [0001] The invention relates to an enterovirus EV71 nucleic acid fluorescence quantitative RT-PCR detection kit and a detection method, which can be applied to laboratory emergency detection of hand-foot-mouth disease and other outbreaks. (2) Background technology [0002] Hand foot mouth disease (HFMD) is a common infectious disease in infants and young children. It is caused by enterovirus. It is clinically characterized by fever, rashes and ulcers on the hands, feet, mouth and other parts. Individual patients can cause myocarditis. , pulmonary edema, aseptic meningoencephalitis and other fatal complications. Hand, foot and mouth disease is a global infectious disease, and there are reports of the prevalence of this disease in most parts of the world. In early March this year, several hospitals in Fuyang, Anhui Province successively admitted and treated children with fever accompanied by rashes in the mouth, hands, feet and buttocks. Laboratory tests c...

AI Technical Summary

Problems solved by technology

[0003] Since HFMD is caused by a variety of human enterovirus infections, among which EV 71 and Cox Al6 are the most common types, in order to quickly find out the cause of the disease and take timely control measures, rapid laboratory diagnosis is urgently needed, but enterovirus traditional The current detection method is virus isolation and neutralization method, which is cumbersome and time-consuming, and is not suitable for early diagnosis

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