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Diarrhea virus detection kit and method

A detection method, a technology for diarrhea virus, which is applied in the field of biotechnology and medical detection, can solve the problems of long detection cycle and low accuracy, and achieve the effect of simple operation, high detection efficiency and improved detection specificity

Active Publication Date: 2013-07-24
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the embodiments of the present invention is to provide a diarrhea virus detection kit and its detection method, aiming to solve the problems of low accuracy and long detection cycle in the existing detection technology

Method used

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  • Diarrhea virus detection kit and method
  • Diarrhea virus detection kit and method

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Embodiment 1

[0081] 1. Preparation of PCR reaction solution

[0082] (1) Synthesize primers and probes according to the following sequence, in which each probe sequence is connected with a fluorescent group at the 5' end and a quencher group at the 3' end:

[0083] Primers and probes against Sarsovirus type 1:

[0084] SAP1-F: 5'-GCAAGCCCTCACGTAGCGAATGTGTCYTGTGCCATTGAGG-3' (SEQ ID NO: 1),

[0085] SAP1-R: 5'-GCAAGCCCTCACGTAGCGAAGTGTATGGAACATTGTGGCTYTC-3' (SEQ ID NO: 2),

[0086] SAP1-probe: FAM-CGCTGCATCAAYCATATGATATATGTGGCGGCAGCG-DABCYL (SEQ ID NO: 3);

[0087] Primers and probes for Saruvirus types 2 and 4:

[0088] SAP2-F: 5'-GCAAGCCCTCACGTAGCGAAATGACCAGGCTCTCGCCAC-3' (SEQ ID NO: 4),

[0089] SAP2-R: 5'-GCAAGCCCTCACGTAGCGAATGGTCCAATGGTGTCCTGDG-3' (SEQ ID NO: 5),

[0090] SAP2-probe: FAM-CCCTCCACYACCAAAATTAGTGTTTGAAATGGAGGG-DABCYL (SEQ ID NO: 6);

[0091] Primers and probes for norovirus type 1:

[0092] NV1-F: 5'-GCAAGCCCTCACGTAGCGAACGCTGGATGCGNTTCCAT-3' (SEQ ID NO: 7),

[0093]...

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Abstract

The invention is applicable to the technical field of biotechnology and medical detection and provides a diarrhea virus detection kit and method. The detection kit comprises PCR (polymerase chain reaction) liquid which comprises a PCR buffer solution, MgCl2, dNTP, DNA polymerase, primer pairs, probe sequences and Homo-tag, wherein the primer pairs and the probe sequences respectively aim at type I, type II and type IV of sapovirus, type I and type II of norovirus, rotavirus, adenovirus and astrovirus, the 5' ends of the probe sequences are connected with fluorophore, and the 3' ends of the probe sequences are connected with quencher. The detection kit provided by the invention can detect seven viruses (subtype) related to diarrhea at one time, and compared with a detection method in the prior art, the detection method provided by the invention has the advantages that operation is simple and convenient, a result is quick to read, and false positive is avoided.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and medical detection, and in particular relates to a diarrhea virus detection kit and a detection method thereof. Background technique [0002] Among the infectious diseases that endanger human health, the incidence of infectious diarrhea ranks third. Infectious diarrhea broadly refers to a group of intestinal infectious diseases caused by various pathogens (except cholera, bacillary and amoebic dysentery, typhoid fever and paratyphoid bacteria), with diarrhea as the main clinical feature. Acute onset, nausea, vomiting, abdominal pain, diarrhea, watery or loose stools, fever and general malaise may also occur. The course of the disease is short and the mortality rate is low. There are three main types of pathogens that cause infectious diarrhea: bacteria, viruses, and parasites. In my country, bacteria and viruses are the main types. Bacterial diarrhea is mainly caused by Shigella, entero...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 扈庆华姜伊祥石晓路李迎慧张海龙林一曼邱亚群
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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