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Diarrhea-related virus multi-gene detection system, and kit and application thereof

A genetic detection and system 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 control the digestive flora of diarrhea in time, and cannot accurately identify various pathogens, etc., to shorten the detection time and cost Low cost and good convenience

Active Publication Date: 2017-09-15
HUADONG HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current conventional 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 empirical treatment in clinical practice , causing problems such as low curative effect, failure to control diarrhea and digestive flora disorder in time
[0007] In summary, current methods for the detection and diagnosis of diarrheal pathogens cannot meet the clinical needs, and the development of new technologies is urgently needed

Method used

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  • Diarrhea-related virus multi-gene detection system, and kit and application thereof
  • Diarrhea-related virus multi-gene detection system, and kit and application thereof
  • Diarrhea-related virus multi-gene detection system, and kit and application thereof

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Embodiment 1

[0043] 1. The composition of the kit

[0044] 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.

[0045] PCR buffer, hot-start DNA polymerase and reverse transcriptase mix were all from Qiagen (Catalog No.: 210212).

[0046] 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.

[0047] The positive control solution is a plasmid mix that includes all gene targets of interest.

[0048] The negative control solution was nuclease-free ultrapure water.

[0049] The primer mixture includes: the nucleotide sequence of the forward primer and the n...

Embodiment 2

[0099] The rest of the diarrhea pathogen multiple gene detection kit of the present embodiment is the same as that of Example 1, except that the primer mixture only includes: respectively targeting human astrovirus, norovirus II, and human enteric adenovirus , rotavirus A, rotavirus B and rotavirus C detection forward and reverse PCR amplification primers, and forward and reverse PCR amplification primers for detection of human RNA internal reference, human DNA internal reference and system quality control internal reference . It is used for the simultaneous detection of 6 infectious diarrhea-associated viruses.

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Abstract

The invention relates to a diarrhea-related virus multi-gene detection system, and a kit and an application thereof. The detection system comprises nine pairs of primers, and the nine pairs of primers include six pairs of diarrhea virus primers, two pairs of human genome internal reference primers and one pair of system quality control internal reference primers. Diarrhea pathogens are directed against human astrovirus, norovirus II, human intestinal adenovirus and rotavirus. The diarrhea pathogen multiple detection system and the kit thereof can realize the simultaneous detection and analysis of multiple diarrhea-related viruses of a night soil sample in a same reaction system without adopting routine detection, so the deficiencies of low flux and long time of routine detection methods are made up, complete, accurate and cheap etiologic diagnosis is clinically provided the first time, and an important reference is improved for individualized medication and accurate medical care.

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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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q2600/16C12Q2600/166
Inventor 张艳梅赵虎吴勇陈敏杨丽华保志军李敏杨长青李冬陈洁
Owner HUADONG HOSPITAL
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