Primer and probe combination and kit for detecting 12 kinds of children digestive tract infection related bacteria

A technology of digestive tract infection and primer probe, applied in the directions of microorganism-based methods, microorganisms, recombinant DNA technology, etc., to achieve the effect of prevention and treatment assistance, high accuracy and high sensitivity

Active Publication Date: 2021-03-16
爱科睿特生物医疗科技(南京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, there is no product that can perform simultaneous PCR detection of the above-mentioned bacterial pathogens of digestive tract infections in children, and an effective method to ensure the accuracy and specificity of the detection

Method used

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  • Primer and probe combination and kit for detecting 12 kinds of children digestive tract infection related bacteria
  • Primer and probe combination and kit for detecting 12 kinds of children digestive tract infection related bacteria
  • Primer and probe combination and kit for detecting 12 kinds of children digestive tract infection related bacteria

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Experimental program
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Embodiment Construction

[0030] Use bacterial positive control as a sample. The kit is composed of primer probe mixed solution, RT-PCR Buffer, mixed enzyme, yang-PCR, and no RNase water; RT-PCR Buffer includes PCR buffer, DATP, DUTP, DCTP, DGTP, and MGCL 2 The mixed enzyme solution includes a HOT START TAQ enzyme, inverse transcriptase, and UNG enzymes; a plasmid containing the amplified gene sequence corresponding to the digestive pathogens in the yang nature control.

[0031] The primer in the amplification system was 100-1000 nm; the final concentration of the probe was 50-500 nm in the amplification system.

[0032] The reactant system of the kit is 30 μL, specifically: a nucleic acid template 7 μL, RT-PCR Buffer 20 μL, 1 μl of mixed enzyme solution and 2 μl of the primer probe mixture.

[0033] Operation and results of the kit

[0034] (1) Extract the sample DNA to the template (extracted from the child's feces), no RNase water, and 7 μL of the yang-nature control each additionally PCR reaction tube,...

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Abstract

The invention relates to a primer group, a probe group and a kit containing the primer group and the probe group, which are used for synchronously detecting various common digestive tract infection related bacteria of children and can be used for simultaneously detecting various common bacteria causing digestive system infection by one-time reaction. The kit is the most comprehensive children digestive tract bacterial infection detection method at present, is high in efficiency and specificity, and provides an auxiliary diagnosis basis for clinical examination. According to the technical scheme, the technical problem that in the prior art, diagnosis and research on bacterial pathogenic bacteria related to children digestive tract infection are still insufficient can be solved.

Description

Technical field [0001] The invention belongs to the field of pathogenic microbial molecular biology, and in particular to primer probe combinations and kits for detection and distinguishing between children with different bacterial pathogens in children with differential tract infections. Background technique [0002] Digestive tract infection is a common disease. It is especially more common in clinical pediatrics. The incidence rate has increased year by year, second only to respiratory system infection, the second largest number of transients, multi-cause bacteria, viruses, fungi or parasite, etc. Pathogenic bacteria infection leads. The annual average canopy, common in bacteria is the jejuno curved bacteria (CJE), CDIFF (CDIFF), murine injury (STYP), vialoxicuris (VP), small intestinal colitis Yerssen ( YENT), adhesive Escherichia coli (EAEC), ETEC, pathogenic large intestine EPEC, hemorrhagic large intestine EEHCE (EEC), invasive large intestine Epsi (EEC), Chikiochemistry (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/10C12Q1/04C12N15/11C12R1/19C12R1/145C12R1/01C12R1/42C12R1/63
CPCC12Q1/689C12Q2600/16
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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