Diarrhea pathogen multi-gene detection system as well as kit and application thereof

A gene detection and pathogen technology, applied in the biological field, can solve the problems that the detection and diagnosis methods of diarrhea pathogens cannot meet the clinical needs, cannot accurately identify multiple pathogens, and cannot diagnose the basis of pathogens, etc., to achieve shortened detection time, low cost, and convenience good sex effect

Active Publication Date: 2017-10-13
HUADONG HOSPITAL +1
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AI Technical Summary

Problems solved by technology

However, the current routine detection methods have the disadvantages of low detection rate, long time-consuming, especially the inability to accurately identify multiple pathogens at the same time, and cannot provide timely, comprehensive and accurate pathogen diagnosis basis for clinical practice, leading to the widespread use of broad-spectrum Empirical treatme

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  • Diarrhea pathogen multi-gene detection system as well as kit and application thereof
  • Diarrhea pathogen multi-gene detection system as well as kit and application thereof
  • Diarrhea pathogen multi-gene detection system as well as kit and application thereof

Examples

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Example Embodiment

[0058] Example

[0059] 1. The composition of the kit

[0060] The diarrhea pathogen multiple gene detection kit of the present embodiment detection kit comprises: primer mixture, PCR buffer (5×PCR Buffer), MgCl 2 solution, dNTPs, hot-start DNA polymerase and reverse transcriptase mix, UDG enzyme, positive and negative controls.

[0061] PCR buffer, hot-start DNA polymerase and reverse transcriptase mix were all from Qiagen (Catalog No.: 210212).

[0062] The hot-start DNA polymerase and reverse transcriptase are mixed together to form a mixed solution, wherein the concentration of the hot-start DNA polymerase in the reaction system is 0.1U / μL, and the concentration of the reverse transcriptase in the mixed solution is 0.1U / μL.

[0063] The positive control solution is a plasmid mix that includes all gene targets of interest.

[0064] The negative control solution was nuclease-free ultrapure water.

[0065] The primer mixture includes: the nucleotide sequence of the forwar...

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Abstract

The invention relates to a diarrhea pathogen multi-gene detection system as well as a kit and application thereof. The detection system comprises 22 pairs of primers, wherein the 22 pairs of primers comprise 19 pairs of diarrhea pathogens, 2 pairs of human genome internal references and 1 pair of system quality control internal reference primers. The diarrhea pathogens are used for treating campylobacter jejuni, shigella, clostridium difficile, salmonella enteritidis, salmonella typhimurium, enterotoxin of escherichia coli, escherichia coli O157, vibrio, yersinia enterocolitica, human astrovirus, norovirus II, human enteric adenovirus, rotavirus and the like. According to the diarrhea pathogen multi-gene detection system and the kit thereof, steps including conventional culture and the like do not need to be adopted and synchronous detection and analysis of a plurality of types of the diarrhea pathogens can be directly carried out on an excrement sample in the same reaction system; the disadvantages of a conventional detection method that the flux is low, the consumed time is long, the detection rate is low and the like are made up; comprehensive, precision and low-cost pathogen diagnosis is provided for clinic at the first time and important reference is provided for individualized drug administration and precision medical treatment.

Description

technical field [0001] The invention relates to a multiple gene detection product and a detection system used in the product, belonging to the field of biotechnology. Background technique [0002] Infectious diarrhea is an intestinal infectious disease caused by a variety of pathogens, and is one of the most common digestive system infectious diseases worldwide. There are about 2 billion diarrheal diseases in the world every year, and about 800 million people suffer from diarrhea every year in my country. Infectious diarrhea has become the second largest infectious disease affecting human health after respiratory tract infection. [0003] Etiological diagnosis is of great significance to the prevention and treatment of infectious diarrhea. "World Organization of Gastroenterology Global Guidelines - Acute Diarrhea in Adults and Children: Global Perspectives (2012)" and "Expert Consensus on the Diagnosis and Treatment of Acute Infectious Diarrhea in Chinese Adults (2013)" cl...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/10
CPCC12Q1/689C12Q1/701C12Q2600/16C12Q2600/166Y02A50/30
Inventor 张艳梅赵虎保志军李敏杨长青陈敏杨丽华李冬吴勇陈洁戴兆云王诗雯杨峰张景皓赵付菊方毅
Owner HUADONG HOSPITAL
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