Method for qualitatively detecting HLA-B*1502 gene with PCR-SSP method and clinical kit

Abstract

The invention belongs to the field of biotechnology, and particularly relates to a method for qualitatively detecting HLA-B*1502 gene with a PCR-SSP method and a clinical detection kit. The method comprises the following steps of: finding 6 specific areas capable of effectively identifying HLA-B*1502 allele type; designing 6 specific primers covering the 6 specific areas, and screening out a high-specificity primer pair applicable to PCR-SSP; and adding dUTP and UDG enzyme (uracil-DNA glycosylase) into a reaction system to solve the problem of PCR cross pollution and further improve the detection reliability. An HLA-B*1502 quick and convenient qualitative detection kit is researched and developed accordingly. The method and the clinical detection kit provided by the invention have the advantages of convenience in operation, short time, strong specificity, high accuracy, low cost and the like, and is suitable for realizing individual safe and reasonable drug use through HLA-B*1502 genotype detection before the Chinese or Asian epileptics take carbamazepine and phenytoin sodium.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for qualitatively detecting HLA-B*1502 gene and a corresponding clinical detection kit. Background technique [0002] Epilepsy is a neurological disease with a high incidence, and the number of cases abroad is 40-70 / 100,000 people every year, and the number in China is about 30 / 100,000 [1] , The main drugs for the treatment of epilepsy include conventional drugs such as phenytoin sodium, carbamazepine, ethosuximide, and sodium valproate, and new drugs such as lamotrigine, oxcarbazepine, and topiramate. Carbamazepine and phenytoin are commonly used first-line antiepileptic drugs at present, and have significant curative effect on complex partial seizure epilepsy. The side effects of antiepileptic drugs include headache, dizziness, unsteady walking, anorexia, nausea, and vomiting. Long-term use of antiepileptic drugs can cause great harm to the memory, movement sp...

AI Technical Summary

Problems solved by technology

At present, the PCR-SSP-based single-specific primer detection kit for HLA-B*1502 in the market is the PG1502 kit [10] from Taiwan’s Pharmigene Company, and its specificity is 98.35% (616 sample), due to the high price, it is inconvenient to promote in the mainland market

In addition, its detection primers cannot distinguish HLA-B*15:13, 15:31, 15:55, 15:88, 15:89, 18:20, 95:12, 95:21, 95:44, 95:70 Waiting for HLA-B alleles, will produce false positives

Moreover, the kit does not take measures to prevent PCR pollution, and it is also easy to cause false positives in clinical testing.

False-positive results prevent patients from choosing the right drug, with consequences particularly severe for children with epilepsy

Compared with other drugs, carbamazepine has less side effects in children, sodium valproate can cause major damage to the liver, and phenobarbital can cause mental decline in children

If there are false positives in the test, children will not be able to choose carbamazepine correctly and take other drugs, causing possible damage

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