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Method for inspecting infectious gene MBL related to SARS CoV infection and MBL infectious gene

A genotype and gene technology, applied in the field of individual susceptibility and its genetics detection, can solve the problems of differences in susceptibility to SARS-CoV infection, affecting the expression level of MBL, etc.

Inactive Publication Date: 2007-01-17
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
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  • Application Information

AI Technical Summary

Problems solved by technology

Polymorphisms in the MBL gene affect MBL expression levels in serum, leading to genotype-dependent differences in susceptibility to SARS-CoV infection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] This embodiment designs and synthesizes 2 pairs of primers, and on the basis of this primer pair, designs a kind of G+230A site that detects the MBL gene associated with SARS-CoV infection susceptibility, and by G-550C (H / L), G-221C(Y / X) and G+230A(A / B), the susceptible haplotype combination LXA / LXA, HYA / LYB, LYA / LYB, LXA / LYB Or the LYB / LYB approach.

[0045] 1. Genomic DNA extraction

[0046] Genomic DNA of peripheral blood leukocytes was extracted by Miller's modified salting-out method, which is conventional in the field.

[0047] 1) Take sodium citrate or EDTA-Na 2 5ml of anticoagulated whole blood (try not to use heparin for anticoagulation), put in a 50ml capped centrifuge tube;

[0048] 2) Add 3-5 times the volume of cold distilled water, invert and mix repeatedly, place in ice for 5 minutes, and centrifuge at 2000rpm at 4°C for 20 minutes;

[0049] 3) Slowly decant the supernatant, add pre-cooled 0.1% Triton-X100 ((volume as above) to the precipitate, gentl...

Embodiment 2

[0061] This embodiment uses the method that embodiment 1 sets up, to the G-550C (H / L), G-221C (Y / X) and G+230A (A) of the MBL gene of 352 routine SARS patients, 392 routine healthy control individuals / B) The genotypes of these 3 loci were analyzed.

[0062] 1. Test object:

[0063] From May 2003 to July 2003, 352 SARS patients meeting the World Health Organization standards were collected from the wards of Beijing Xiaotangshan Hospital. Among the 352 patients, 52 patients had some basic diseases that may increase the risk of SARS, such as diabetes, hypertension, heart disease, tuberculosis, asthma and tumors and other important diseases (for PI group); another 20 were patients with severe SARS who were admitted to the intensive care unit (ICU) or died (for P II group); the remaining 280 cases were P III Group. Another 392 healthy individuals matched in age, gender and race were collected as the control group. The sample collection process was carried out in accordance wit...

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Abstract

The present invention relates to the PCR product sequencing assay of mannose-binding lectin gene (MBL) correlative with the susceptibility of SARS-CoV infection. The present invention also relates to the assay of the haplotypes correlative with the susceptibility of SARS-CoV infection that is composed of three single nucleotide polymorphism sites(SNPs): MBL gene G-550C,G-221C and G+230A. The said assay is to determine the genotype of the three SNPs by sequencing the PCR product first and then to calculate the haplotypes with PHASE programme. The present invention further relates to the detection kit of the above SNPs and the usage. Furthermore, the present invention combines the G+230A polymorphism sites and the haplotype composed of G-550C,G-221C and G+230 with SARS-CoV susceptibility, confirms that MBL is a new susceptibility gene included in SARS-CoV infection, and provides new genetic counseling content for the prevention and control of SARS-CoV infection of the population.

Description

technical field [0001] The invention relates to the individual susceptibility and genetic detection method in the prevention of severe acute respiratory syndrome (Severe Acute Respiratory Syndrome, SARS) population. Specifically, the present invention relates to a genotype detection method of the susceptibility gene mannose-binding lectin (mannose-binding lectin, MBL) gene associated with SARS coronavirus (SARS Coronavirus, SARS-CoV) infection. The present invention further relates to a detection kit for the above-mentioned susceptibility gene genotype, and the application of the kit. In addition, the present invention also relates to the application of the above detection method and susceptibility gene in the prevention and control of SARS and genetic counseling. Background technique [0002] SARS is a newly identified human infectious disease caused by a novel coronavirus, SARS-CoV. As of the end of June 2003, more than 8,000 SARS cases were reported in more than 25 coun...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 贺福初周钢桥张红星翟芸智联腾杨灏
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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