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

A detection kit, RT-PCR technology, used in fluorescence/phosphorescence, microbial determination/inspection, biochemical equipment and methods, etc., can solve problems such as cumbersome and time-consuming, not suitable for early diagnosis

Inactive Publication Date: 2008-12-24
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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  • Abstract
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  • Claims
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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, ra...

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

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Embodiment 1

[0041] 1 Materials and methods

[0042] 1.1 Virus strains and clinical specimens:

[0043] Enterovirus EV71, Coxsackie group A (CA) CA4, CA16, CA21, Coxsackie B5, E6, E30, POLIO type 1 and other virus strains were obtained from Chinese Academy of Medical Sciences, Beijing Institute of Biological Products and Zhejiang Province Disease Prevention Control center isolates. The clinical samples come from the cerebrospinal fluid, herpes fluid, feces, etc. of the suspected patients of the recent outbreak of hand, foot and mouth disease in Zhejiang Province. After the samples are collected, they are transported to the laboratory with ice.

[0044] 1.2 Primers and probes

[0045] Download the enterovirus EV71 gene sequences of different years and regions from GenBank, use biological software for homology comparison, and design specific primers and TaqMan probes in the VP1 region. The sequence is:

[0046] EV71YG F1: 5'-TGATTGAGACACGC / GTGTGTT / CCTTA-3',

[0047] EV71YG R1: 5'-CCC GCT...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 卢亦愚严菊英徐昌平冯燕陈寅
Owner ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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