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Primer and probe combination and kit for detecting 9 children digestive tract pathogens

A primer probe, pathogen technology, applied in the field of molecular biology of pathogenic microorganisms, to achieve the effect of strong 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

[0009] At present, there is no product that can perform simultaneous PCR detection of the above-mentioned multiple digestive tract infection virus pathogens 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 9 children digestive tract pathogens
  • Primer and probe combination and kit for detecting 9 children digestive tract pathogens
  • Primer and probe combination and kit for detecting 9 children digestive tract pathogens

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Experimental program
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Embodiment Construction

[0028] Use a virus positive control as a sample. The kit consists of primer-probe mixture, RT-PCR buffer, mixed enzyme solution, positive quality control and RNase-free water; RT-PCR buffer includes PCR buffer, dATP, dUTP, dCTP, dGTP and MgCl 2 ; The mixed enzyme solution includes Hot Start Taq enzyme, reverse transcriptase and UNG enzyme; the positive quality control contains the plasmid or RAN particle corresponding to the amplified gene sequence of the gastrointestinal pathogen.

[0029] 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.

[0030] 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 mixture.

[0031] Kit operation and result determination:

[0032] (1) Add 7 μL each of the sample DNA / RNA co-extraction template (extracted from the stool of ...

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Abstract

The invention relates to a primer and probe combination and kit for detecting 9 children digestive tract pathogens. A primer group, a probe group and the kit can realize simultaneous detection of a plurality of common viruses causing digestive system infection by one reaction, and a method is the most comprehensive detection method for children digestive tract virus infection at present, and is high in efficiency and strong in specificity, 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 virus pathogens related to children digestive tract infection 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 multiple fluorescence quantitative PCR kit for detecting and distinguishing nucleic acid series of children's digestive tract infection virus, and a detection method of the kit. 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. It can occur throughout the year, and the most common viruses are adenovirus (AdV), astrovirus (AsV), norovirus (NV), rotavirus (RV), sacrovirus (SaV), cytomegalovirus Virus (CMV), Coxsackie virus A (CoxA16), enterovirus 71 (EV71), human coronavirus (Hcov), etc. The symptoms caused by ga...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/686C12Q1/701C12Q1/705C12Q2600/16C12Q2537/143C12Q2561/101C12Q2521/107C12Q2563/107
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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