Kit, primer probe composition and method for jointly detecting multiple enteroviruses

A technology for joint detection and enterovirus, applied in the field of rotavirus and human astrovirus kits, Zaru virus, and norovirus joint detection, which can solve the problem of low specificity and achieve high sensitivity and good specificity. Effects of Sex and Sensitivity

Pending Publication Date: 2022-04-08
HEMOSMART MEDICAL TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some literatures provide protocols capable of detecting multiple enteroviruses, the specificity is low

Method used

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  • Kit, primer probe composition and method for jointly detecting multiple enteroviruses
  • Kit, primer probe composition and method for jointly detecting multiple enteroviruses
  • Kit, primer probe composition and method for jointly detecting multiple enteroviruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: Kit

[0063] The kit is a kit for simultaneously detecting four enteroviruses, including nucleic acid reaction solution A and nucleic acid reaction solution B. Nucleic acid reaction solution A and RT-qPCR amplification reaction solution constitute a triple detection system for norovirus / zarovirus / human internal reference; nucleic acid reaction solution B and RT-qPCR amplification reaction solution constitute rotavirus / astrovirus / Human-derived internal reference triple detection system. The RT-qPCR amplification reaction solution was purchased from Beijing Tiangen Biotechnology Co., Ltd., and specifically included the following components: Taq enzyme, buffer, and dNTPs.

[0064] The specific composition of nucleic acid reaction solution A includes:

[0065] (1) Forward primer F1, reverse primer R1 and probe P1 of Norovirus GI group, the 5' end of the probe P1 is labeled with a FAM fluorescent reporter group, and the 3' end is labeled with a BHQ1 fluores...

Embodiment 2

[0091] Embodiment 2: the detection pseudovirus result of kit of the present invention

[0092] The pseudoviruses of Norovirus (NV), Sarovirus (SaV), Rotavirus (RV) and Human Astrovirus (HAstVs) were detected using the kit of Example 1. The pseudoviruses of the above four viruses were serially diluted, and the gradients were 5×10^6 copies / mL, 5×10^5 copies / mL, 5×10^4 copies / mL, 5×10^3 copies / mL, 5×10^2 copies / mL, test the sensitivity of the kit. For test results, see Figure 1 to Figure 4 As shown, the results show that the sensitivity of Norovirus can reach 500 copies / mL, and the sensitivity of Zarovirus can reach 500 copies / mL. Rotavirus can reach 500 copies / mL, and astrovirus can reach 500 copies / mL.

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Abstract

The invention discloses a kit, a primer probe composition and a method for jointly detecting various enteroviruses. The kit comprises a first nucleic acid reaction solution and a second nucleic acid reaction solution, and in the first nucleic acid reaction solution, nucleotide sequences of a primer pair and a probe of a norovirus GI group are sequentially shown as SEQ ID NO.1-3; the nucleotide sequences of the primer pair and the probe of the norovirus G2 group are as shown in SEQ ID NO.4-6 in sequence; the nucleotide sequences of the primer pair and the probe of the fiveleaf akebia virus are shown as SEQ ID NO.7-9 in sequence; in the second nucleic acid reaction liquid, the nucleotide sequences of the primer pair and the probe of the rotavirus group A are shown as SEQ ID NO.10-12 in sequence; the nucleotide sequences of the primer pair and the probe of the rotavirus B group are sequentially shown as SEQ ID NO.13 to SEQ ID NO.15; the nucleotide sequences of the primer pair and the probe of the astrovirus are as shown in SEQ ID NO. 16-18 in sequence. The kit can be used for jointly detecting norovirus, fiveleaf virus, rotavirus and human astrovirus, and has relatively high specificity and sensitivity.

Description

technical field [0001] The invention belongs to the field of enterovirus detection kits, and relates to a kit, a primer probe composition and a method for joint detection of multiple enteroviruses, in particular to a joint detection of norovirus, zaravirus, and rotavirus and human astrovirus kits. Background technique [0002] Viral gastroenteritis, also known as viral diarrhea, is a group of acute intestinal infectious diseases caused by a variety of viruses. The clinical features are acute onset, nausea, vomiting, abdominal pain, diarrhea, watery or loose stools, fever and general malaise, etc. The course of the disease is short and the mortality rate is low. The clinical manifestations of gastroenteritis caused by various viruses are basically similar. There are many types of viruses related to acute gastroenteritis, among which the main viruses causing viral diarrhea are Norovirus (NV for short), Rotavirus (RV for short), and Sapovirus (SaV for short). , Human Astrovi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
Inventor 颜菁孙永祥曾毓磊
Owner HEMOSMART MEDICAL TECH LTD
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