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Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by sulfasalazine

A technology of sulfasalazine and leukocyte antigen, which is applied in biological testing, microbial measurement/testing, material inspection products, etc., and can solve problems such as unseen kit reports

Active Publication Date: 2018-09-21
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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Method used

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  • Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by sulfasalazine
  • Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by sulfasalazine
  • Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by sulfasalazine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Collection and extraction of genes

[0047] Sulfasalazine-induced DRESS patients were from Huashan Hospital Affiliated to Fudan University, Shanghai, China.

[0048] The diagnostic criteria were determined by Bocquet et al. (Bocquet, H., M. Bagot and J.C. Roujeau, Drug-induced pseudolymphoma and drug hypersensitivity syndrome (Drug Rash with Eosinophilia and Systemic Symptoms: DRESS). Semin Cutan Med Surg, 1996.15(4): p.250 -7.) proposes that skin rash, hematological abnormal changes (eosinophilia greater than 1500 / mm3 and abnormal lymphocytes) and system involvement symptoms (lymph node enlargement, diameter greater than 2cm or hepatitis transaminase are normal) More than 2 times, interstitial nephritis, interstitial pneumonia or carditis) three conditions were diagnosed as DRESS. Under the premise of informed consent, participants participated in the research of this subject, collected blood, and signed an informed consent form. All enrolled patients were...

Embodiment 2

[0050] Example 2: Detection of HLA genotyping

[0051] The present invention adopts PCR-SSO method (recommended Kit-One Lambda, CA, USA) for HLA genotyping. Its principle is to amplify the polymorphic region of HLA first, carry out isotopic or non-isotopic labeling on the PCR products during the amplification process, and then design a series of oligonucleotide probes for the PCR amplification products according to the principle of base pairing and immobilize them in Finally, the PCR product was hybridized with the probe on the membrane, and autoradiography was used to judge the result according to the signal. HLA genotyping was performed according to the standard steps of the kit. (i.e. DNA samples, substrates, Taq enzymes, primers are mixed and mixed, added to the amplification plate, amplified according to the conditions in the kit instructions, and the amplified products are hybridized, stained and read). Results were analyzed by HLA Fusion software (Onelambda, CA, USA...

Embodiment 3

[0052] Example 3: HLA-B*13:01 is associated with DRESS caused by sulfasalazine

[0053] Statistics: SPSS16.0 was used to calculate Odds Ratio (OR) and its 95% confidence interval. If necessary, Haldane's correction was used, and chi-square test was used for statistical analysis. The statistical significance level was set at P less than 0.05.

[0054] Results: A total of 6 patients with DRESS caused by sulfasalazine were collected (Table 1), and 30 patients were in the sulfasalazine-resistant group. The positive ratio of HLA-B*13:01 in sulfasalazine-induced DRESS patients was significantly different from that in normal control group and sulfasalazine-resistant group (Table 2). HLA-B*13:01 binding model with sulfasalazine see figure 1 (Schematic diagram of the binding model of sulfasalazine to HLA-B*13:01).

[0055] Table 1. Clinical features and HLA types of DRESS induced by sulfasalazine

[0056]

[0057] Table 2. HLA-B*13:01 positive ratio of sulfasalazine-induced DRESS...

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Abstract

The invention provides a human leukocyte antigen gene detection kit for screening skin drug adverse reaction caused by salazosulfapyridine, belongs to the technical field of biomedicine, and relates to a detection kit of human leukocyte antigen gene HLA-B*13:01, wherein the HLA-B*13:01 gene can be adopted as the marker gene of salazosulfapyridine-induced DRESS. According to the present invention, DNA is extracted from the peripheral blood of a patient, and the HLA-B*13:01 gene is detected by using the current method, such as PCR-SSO (sequence-specific oligonucleotide probe) method; and the HLA-B*13:01 gene detection kit of the present invention can be used for screening before the salazosulfapyridine use and screening the target drug for treatment of salazosulfapyridine-induced DRESS, wherein the screening before the salazosulfapyridine use can guide the clinical medication so as to reduce the occurrence of the salazosulfapyridine-induced DRESS, and the drug screening mainly act on the HLA-B*13:01 molecules in a targeting manner so as to block the interaction between the HLA-B*13:01 molecules and the salazosulfapyridine or the metabolites thereof in the pathogenesis.

Description

technical field [0001] The invention belongs to the field of biomedicine and reagent detection, and relates to a human leukocyte antigen gene detection kit. Specifically, it involves the human leukocyte antigen gene--HLA-B*13:01 gene, which is related to the pathogenesis of sulfasalazine-induced skin adverse drug reactions with eosinophilia and systemic symptoms. Background technique [0002] Sulfasalazine is a commonly used clinical drug, which is mainly used for inflammatory bowel disease and rheumatoid arthritis (Mottonen, T., et al., Comparison of combination therapy with single-drug therapy in early rheumatoid arthritis: a randomized trial. FIN-RACo trialgroup.Lancet,1999.353(9164):p.1568-73.Van Hees,P.A.,et al.,Effect of sulphasalazine in patients with active Crohn's disease:a controlled double-blind study.Gut,1981.22(5): p.404-9. Dissanayake, A.S. and S.C. Truelove, A controlled therapeutic trial of long-term maintenance treatment of ulcerative colitis with sulphazal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68
Inventor 邢清和骆肖群徐金华杨凡萍
Owner FUDAN UNIV
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