CN102108382A

CN102108382A - Genetic testing for risk of causing serious adverse reactions of skin of carbamazepine

Info

Publication number: CN102108382A
Application number: CN2009102004862A
Authority: CN
Inventors: 冯哲民; 邹祖烨; 贺宪民
Original assignee: SHANGHAI ZHUJIAN BIOENGINEERING CO Ltd
Current assignee: SHANGHAI ZHUJIAN BIOENGINEERING CO Ltd
Priority date: 2009-12-23
Filing date: 2009-12-23
Publication date: 2011-06-29

Abstract

The invention discloses a genetic testing method for risk of causing serious adverse reactions of carbamazepine. The genetic testing method comprises the following steps: collecting oral mucosa cells of a subject, extracting genome DNA (deoxyribonucleic acid) of the oral mucosa cells, testing HLA (human leukocyte antigen)-B*1502 allelotype of the genome DNA and evaluating the risk of causing the serious adverse reactions of the skin of an anti-epileptic medicament-carbamazepine, thereby providing reference basis for clinical individual medication.

Description

Carbamzepine causes skin serious adverse reaction Genetic Detection

Technical field

The present invention relates to molecular biology and medical field, more specifically, the present invention relates to a kind of method that causes skin serious adverse reaction risk by Genetic Detection control Carbamzepine.

Background technology

Epilepsy is a kind of disease and syndrome, is feature with the intermittent central nervous system function imbalance due to the unexpected repeatedly over-discharge can of brain neuroblastoma unit.In the treatment of epileptic seizures, antiepileptic drug has special important meaning.(Carbamazepine is a clinical line antiepileptic drug commonly used CBZ) to Carbamzepine, is used for the treatment of oromaxillo-facial region trigeminal neuralgia, glossopharyngeal neuralgia and epileptic seizures.But CBZ in use easily produces various toxic side effect, even threat to life, wherein skin lesion commonly, the about 10% patient Yi Fasheng fash of taking for a long time, eczema, urticaria, dermatomyositis, allergic dermatitis, exfoliative dermatitis, toxic epidermal necrolysis (Toxic epidermal necrolysis, TEN), hebra's disease, lupus erythematosus syndrome and Stevens-Johnson syndrome (SJS) etc., the probability of occurrence of serious skin reaction is approximately 1/10000, lethality rate reaches more than 30%, and wherein the lethality rate of SJS/TEN is up to 40%.Therefore, the clinician should be prudent especially before using CBZ, assessing use rationally on its basis that causes the skin adverse reaction risk.

Human leucocyte antigen (human leukocyte antigen, HLA) system, promptly human main (the majorhistocompatibility complex of histocompatibility complex, MHC), be positioned at human No. 6 the short arm of a chromosome 6p21.31, form the about 3600kb of dna fragmentation length by the closely linked multiple allelomorphos of a group site.The HLA complex structure is very complicated, be up to now known to the most complicated genetic polymorphism system, starting and participating in playing a significant role in the identification of body specific immunity, the immunne response.The HLA complex body has identified 224 locus, and functioning gene that wherein can expression product is 128, and be divided into 3 districts by the arrangement of HLA complex body on karyomit(e) traditionally: I genoid district is positioned at the HLA complex body away from kinetochore one end; II genoid district is positioned at the nearly kinetochore of HLA complex body one end; III genoid district is positioned between the two.3 gene regions contain the dozens of locus, are called HLA-I, HLA-II, HLA-III genoid (Fig. 2).The I class comprises A, B, the classical gene of C and E, F, G, X, J, H nonclassical gene.HLA-B*1502 is 1 gene hypotype of position, HLA gene Building B.To existing more than 1000 kind of allelotrope in HLA-B seat in 2009, its polymorphism mainly was positioned at the 2nd, No. 3 exon of HLA-B gene.There is strong correlation in the Stevens-Johnson syndrome/toxic epidermal necrolysis disease (CBZ-SJS/TEN) that has report HLA-B*1502 and CBZ to cause, and is the strongest in the dependency about HLA mark and a certain disease so far.

Studies show that the HLA-B*1502 positive patient takes the risk significance that SJS/TEN appears in CBZ and strengthen, if US Basic Application Number research and development report FDA suggestion patient has the popular geographic blood lineage of HLA-B*1502, should carry out the HLA-B*1502 genotype detection before taking CBZ first, to lower the high risk of antiepileptic drug.There is no the dependency report of relevant HLA-B*1502 gene and CBZ-SJS/TEN at present in CONTINENTAL AREA OF CHINA.Therefore, carry out the research of this aspect, can further improve skin-type adverse drug reaction and HLA-B*1502 correlation research that CBZ causes, thereby provide reference frame for clinical individual medication.

Summary of the invention

The invention provides a kind of method that causes skin serious adverse reaction risk by Genetic Detection control Carbamzepine.

By gathering person under inspection's oral mucosa cell, extract its genomic dna, HLA-B*1502 allelotype to its genomic dna detects, and causes the risk of skin serious adverse reaction from gene aspect assessment antiepileptic drug Carbamzepine, thereby provides reference frame for clinical individual medication.

Embodiment

Below in conjunction with specific embodiment, further set forth the present invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, or the condition of advising according to manufacturer.

The use of embodiment detection kit

The extraction of step 1:DNA template

Genomic dna with silica gel adsorption extracting mouth epithelial cells.

Step 2: quantitative fluorescent PCR reaction

Use the quantitative fluorescent PCR suit in the detection kit, carry out the quantitative fluorescent PCR reaction, the system of reaction is cumulative volume 10 μ l, and comprising concentration is dna profiling 2 μ l, 1 μ l 10X quantitative fluorescent PCR reaction buffer, 0.1 μ l 25mM dNTP mixed solution, the 0.6 μ l 25mM MgCl of 20ng/ μ l ₂The band VIC fluorescent probe that adopted primer and antisense primer each 0.225 μ l, 10 μ M are arranged of solution, 0.025 μ l (5units/ μ l) Taq archaeal dna polymerase, 20 μ M and each 0.25 μ l of band FAM fluorescent probe, deionized water 5.325 μ l.

React on the pcr amplification instrument, reaction conditions is 50 ℃, 2 minutes, 95 ℃, 10 minutes, carries out 95 ℃ of 60 round-robin, 30 seconds, 60 ℃, 1 minute.Reaction finishes the back and read the fluorescent amount on quantitative real time PCR Instrument.

Step 3: gene type assay

According to the gene type diagram that the test kit working instructions indicate gene type assay is carried out in the SNP site.

The those skilled in the art that are familiar with fluorescent quantitative PCR technique can be by the final sample fluorescence volume that shows on the identification quantitative real time PCR Instrument, can determine the genotype in the SNP site detected according to the power of different sequence fluorescence probe signals.

Claims

1. a Carbamzepine causes skin serious adverse reaction Genetic Detection method, it is characterized in that: the oral mucosa cell of gathering the person under inspection, extract its genomic dna, HLA-B*1502 allelotype to its genomic dna detects, cause the risk of skin serious adverse reaction from gene aspect assessment antiepileptic drug Carbamzepine, thereby provide reference frame for clinical individual medication.