用焦磷酸微测序技术进行HLADRB基因型分析
焦磷酸微测序,,多态现象(遗传学);HLA抗原;焦磷酸微测序;基因型
A pyrosequencingbased method for high resolution HLADRB genotypingYUAN JianLin, WU GuoJun, XUE Li, ZHAO JinRong, BAI YuJie, YANG Fang2, WANG He, ZHANG YunTao, YANG LiJun
(1Department of Urology, Xijing Hospital, 2Institute of Gene Diagnosis, Fourth Military Medical University, Xian 710033, China)
【Abstract】 AIM: To develop a pyrosequencingbased typing (PSBT) approach for high resolution identification of HLADRB alleles. METHODS: The DNA fragments from HLADRB exon2 were obtained using allelespecific or multiplex PCR amplification. The PSBT of purified DNA was performed. RESULTS: The polymorphic residues of HLADRB genes were identified in each pyrosequencing reaction and read length up to 90 nucleotides was obtained. A blood sample containing heterozygous alleles (DRB10405 and DRB301011) was analyzed using purified DNA respectively from allelespecific or multiplex PCR reaction, the results of which were consistent with those from the traditional method. CONCLUSION: Pyrosequencing used in the analysis of HLADRB alleles has the advantage of high resolution and can be widely employed clinically in donor/recipient selection. ......
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