Specific primer probe combination for detecting HLA-A*24:02 allelic genes and kit and detection method

Abstract

The invention discloses specific primer probe combinations for detecting HLA-A*24:02 allelic genes and a kit and a detection method. Specifically, on the basis of a TaqMan probe detection method, twopairs of primer probe combinations for highly specifically amplifying HLA-A*24:02 allelic genes are designed, namely an upstream primer Fp1: 5'-GGTTCTC ACACCCTCCA GATGATGGT-3', a downstream primer Rp1: 5'-CTCCAGGTATCTGCGGAGCACG-3', a probe 1: 5'-FAM-CCACTTGCGCTTGGTGATCTGAGCC-BHQ2-3', an upstream primer Fp2: 5'-CTCGTCCC CAG GCTCCCTC-3', a downstream primer Rp2: 5'-CACCGGCCTCGCTCTGGT-3', a probe 2:5'-CY5-GAAGGCCCACTCACAGACTGACCGAG-BHQ2-3'; FAM is 6-carboxyfluorescein; Cy5 is Cyanine 5; BHQ2 is Black Hole Quencher-2; the two specific primers and probes together with reference gene primers and probes are put into a same tube to implement a multiple fluorescent PCR (Polymerase Chain Reaction), and results are analyzed through fluorescent amplification curves. The specific primer probe combinations have the characteristics of being good in specificity, flexible and rapid, high in flux, free of pollution, high resolution rate, possible in real-time reaction process monitoring, and the like,and are applicable to detection on whole genome DNA (Deoxyribonucleic Acid) samples HLA-A*24:02 in human peripheral blood and saliva.

Description

technical field [0001] The invention belongs to the fields of pharmacogenomics and gene diagnosis, and in particular relates to a specific primer-probe combination for detecting HLA-A*24:02 alleles. Background technique [0002] Carbamazepine (CBZ), lamotrigine (LTG), and phenytoin (PHT) are a class of structurally similar aromatic antiepileptic drugs (AEDs) commonly used clinically, and according to statistics, about 1% of the world's population uses them . AEDs are the most common drugs that are likely to cause drug eruption reactions. The incidence rates of drug eruptions caused by CBZ, LTG, and PHT were 3.7%, 4.8%, and 5.9%, respectively. Severe drug eruption reactions such as Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN), with a fatality rate of up to 20-50% [1,2] . In recent years, studies have found that drug eruption reactions caused by aromatic antiepileptic drugs are closely related to individual genetic susceptibility, among which the gene...

AI Technical Summary

Problems solved by technology

[0006] However, since there are more than 3,000 known subtypes of HLA-A alleles, and the sequences of HLA alleles have strong diversity and high homology, it is difficult to obtain them simply by using primer design software. Specific primer and probe sets for HLA-A*24:02 gene

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