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H7N9 avian influenza illness probability detection kit

A detection kit, avian influenza technology, applied in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc., can solve problems such as systemic inflammatory response, cytokine storm, and unclear pathogenesis

Active Publication Date: 2014-12-17
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] H7N9 avian influenza virus binds to sialic acid receptors located on the surface of human upper respiratory epithelial cells, and can induce cytokine storm at the same time, leading to systemic inflammatory response, ARDS, shock and multiple organ failure, but the exact pathogenesis is not yet known clear

Method used

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  • H7N9 avian influenza illness probability detection kit
  • H7N9 avian influenza illness probability detection kit
  • H7N9 avian influenza illness probability detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1: Sample preparation and sanger sequencing verification

[0027] Detect 5 genes and 10 loci of 8 H7N9 avian influenza patients respectively, divide these 10 loci into 6 fragments (as shown in Table 2), and design primers for the exon sequence of each fragment , Obtain the sequence of each fragment by PCR amplification, product purification, and sanger sequencing, and compare whether it belongs to the mutant type or the wild type according to the sequence determination results, and verify the correlation between each gene and H7N9 avian influenza. The specific method steps are as follows:

[0028] Step 1: DNA Extraction:

[0029] The peripheral blood of 8 patients with H7N9 avian influenza was collected, and the genomic DNA in the peripheral blood leukocytes was extracted using the sQ Blood DNA Kit II produced by omegabiotek (article number: D0714-250), and the concentration of genomic DNA was detected by NanoDrop2000 and agarose gel electrophoresis and purity...

Embodiment 2

[0038] Example 2: H7N9 Avian Influenza Disease Probability Detection Kit and Preparation Method

[0039] 一种H7N9禽流感患病概率检测试剂盒,包含对于CPT2-chr1:53676448&53676401、CPT2-chr1:53679229&53679028、FCGR2A-chr1:161479745、IFITM3-chr11:320772、RPAIN-chr17:5326145和TLR3-chr4:187004074&187004217这 6 pairs of PCR primers for detection of 9 sites in 5 genes.

[0040] The sequences of 6 pairs of PCR primers for detecting these 9 sites are as follows:

[0041]

[0042] According to the method described in step 1 in embodiment 1, extract the DNA of the person to be tested, use the extracted DNA as a template and the above primers to carry out PCR reaction according to the PCR conditions described in step 2 in embodiment 1, and perform PCR according to conventional methods in the art The product was purified, and the purified product was subjected to sanger sequencing. Observe whether the sequence obtained by sequencing has mutations at these 9 sites. If the mutations of these 9 sites are included, ...

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Abstract

The invention provides an H7N9 avian influenza illness probability detection kit. The H7N9 avian influenza illness probability detection kit is characterized by comprising six pairs of PCR (Polymerase Chain Reaction) primers for detecting nine loci of five genes of CPT2-chr1: 53676448 & 53676401, CPT2-chr1: 53679229 & 53679028, FCGR2A-chr1: 161479745, IFITM3-chr11: 320772, RPAIN-chr17: 5326145 and TLR3-chr4: 187004074 & 187004217. The H7N9 avian influenza illness probability detection kit can find mutations at nine loci in total for an H7N9 patient and is applicable to detection for patients infected by H7N9 avian influenza and high-risk groups, thereby providing a certain value for prevention and control of H7N9 infection epidemic. Virulence genes identified by the H7N9 avian influenza illness probability detection kit may play a role in the pathogenesis of an H7N9 avian influenza infection process by entering epithelial cells in a respiratory tract in H7N9 infection and may also be susceptible genes of H7N9 avian influenza.

Description

technical field [0001] The invention relates to a kit for detecting the prevalence probability of H7N9 avian influenza. Background technique [0002] On March 31, 2013, 3 confirmed cases of human infection with H7N9 avian influenza virus occurred in Shanghai and Anhui, of which 2 died and 1 was critically ill. Since then, the epidemic has spread rapidly. As of June 2013, a total of 132 confirmed cases have been reported nationwide. The patients were mainly characterized by severe pneumonia and acute respiratory distress syndrome (ARDS), which has resulted in 43 deaths. Definite evidence of human-to-human transmission was found. [0003] The H7N9 avian influenza virus belongs to the Orthomyxoviridae influenza A virus, which is a single negative-strand RNA virus with an envelope and segments. Avian influenza viruses can be divided into 16 H subtypes (H1-H16) and 9 N (N1-N9) subtypes based on the antigenicity of their outer membrane hemagglutinin (H) and neuraminidase (N) pro...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12R1/93C12Q1/70
CPCC12Q1/6883C12Q2600/156
Inventor 宋元林周文浩王明帮陈翠翠杨琳卢水华王琴唐薪竣白春学樊晓梅席秀红李涛
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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