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Primer probe composition and kit for synchronously detecting 29 pathogens related to digestive tract infection of children

A technology for the simultaneous detection of digestive tract infections, applied in the direction of recombinant DNA technology, microbial detection/inspection, resistance to vector-borne diseases, etc.

Inactive Publication Date: 2021-03-19
爱科睿特生物医疗科技(南京)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, there is no product that can simultaneously detect bacteria, viruses, fungi, and parasitic pathogens of the above-mentioned children's digestive tract infections, and an effective method to ensure the accuracy and specificity of the detection

Method used

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  • Primer probe composition and kit for synchronously detecting 29 pathogens related to digestive tract infection of children
  • Primer probe composition and kit for synchronously detecting 29 pathogens related to digestive tract infection of children
  • Primer probe composition and kit for synchronously detecting 29 pathogens related to digestive tract infection of children

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Embodiment Construction

[0037] Use positive controls of bacteria, viruses, fungi, and parasitic pathogens as samples. The kit consists of primer-probe mixture, RT-PCRbuffer, mixed enzyme solution, positive quality control and RNase-free water; RT-PCRbuffer includes PCR buffer, dATP, dUTP, dCTP, dGTP and MgCl 2 ; The mixed enzyme solution includes HotStartTaq enzyme, reverse transcriptase and UNG enzyme; the positive quality control contains a plasmid corresponding to the amplified gene sequence of the gastrointestinal pathogen.

[0038] The final concentration of the primer in the amplification system is 100-1000nM; the final concentration of the probe in the amplification system is 50-500nM.

[0039] The reaction system of the kit is 30 μL, specifically: 7 μL of nucleic acid template, 20 μL of RT-PCR buffer, 1 μL of mixed enzyme solution, and 2 μL of primer-probe mixed solution.

[0040] Kit operation and result determination:

[0041] (1) Add 7 μL each of the sample DNA co-extraction template (ex...

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Abstract

The invention relates to a primer probe composition and a kit for synchronously detecting 29 pathogens related to digestive tract infection of children. According to the primer probe composition and the kit, a primer group, a probe group and the kit can realize simultaneous detection of a plurality of common bacteria, viruses, fungi and parasitic pathogens causing digestive system infection by onereaction, the method is the most comprehensive children digestive tract infection detection method at present, the efficiency is high, the specificity is strong, and an auxiliary diagnosis basis is provided for clinical examination. By means of the technical scheme, the technical problem that in the prior art, diagnosis and research on pathogens related to digestive tract infection of children are still insufficient can be solved.

Description

technical field [0001] The invention belongs to the field of molecular biology of pathogenic microorganisms, and in particular relates to a combination of primers and probes and a kit for serial detection and differentiation of bacteria, viruses, fungi and parasite pathogens in children with digestive tract infection. Background technique [0002] Gastrointestinal infection is a common disease, which is especially common in clinical pediatrics. The incidence rate is increasing year by year. It is second only to respiratory system infection and ranks the second most common disease in pediatrics. It is mostly caused by bacteria, viruses, fungi or parasites, etc. Caused by pathogenic infection. Bacteria, viruses, fungi, and parasitic pathogens are common throughout the year, including Campylobacter jejuni (CJE), Clostridium difficile (Cdiff), Salmonella typhimurium (STYP), and Vibrio parahaemolyticus (VP) , Yersinia enterocolitica (Yent), adhesive Escherichia coli (EAEC), toxi...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6893C12Q1/6895C12Q1/70C12Q1/04C12N15/11
CPCC12Q1/689C12Q1/6893C12Q1/6895C12Q1/701C12Q2600/16Y02A50/30
Inventor 董晓艳熊永清王洪
Owner 爱科睿特生物医疗科技(南京)有限公司
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