HLA-DR抗原标准血清学分型与基因分型的比较研究

HLA-DR typing by standard serology and PCR-amplification with sequence-specific primers: A comparative study

目的:比较研究顺序特异引物聚合酶链反应(PCR-SSP)方法与标准血清学方法对人类白细胞抗原-DR(HIA-DR)分型的精确性和临床应用价值.方法:选择肾移植受者101例,供者61例,采用PCR-SSP和血清学方法同步行HLA-DR分裂,比较其检测时间、敏感性、特异性和临床实用性.结果:162份样本,PCR-SSP分型均获成功,检出DR等位基因总数308个,分型时间5小时,特异性和重复性100%.血清学分型耗时20小时,8个位点不肯定,29个分型错误,35个空白位点中20个存在第二个位点,总误差率30.2%.结论:PCR-SSP方法用于HLA-DR分型较血清学方法快速、精确,试剂与临床样本易得,适合于临床应用.

Objective DMA tying for HLA-DR by polymerase chain reacion with sequence-specific primers (PCR-SSP) compared with standard serology to evaluate the reliability and clinical practicability. Methods Double-blind typing for HLA-DR alleles was carried out using DNA typing by PCR-SSP and standard serology by microlymphocytotoxicity technique in 61 donors and 101 recipients of cadeveric transplantation. Matching time, sensitivity, specificity and clinical practicability were compared according to typing results by both methods. Results All 162 samples were able to be typed by PCR-SEP. A total of 308 alleles were detected(16 DR blank). The results of matching were confirmed by analysis with restriction endonucleases and Southem hybridization. The specificity and reproducibility were 100%. HLA-DR typing was performed in 5 hours by PCR-SSP or 20 hours by serology. The discrepancy rate between PCR-SSP and serological HLA-DR typing was 30.296(35.6% for kidney recipients, 21.3% for donors). The discrepancies consisted of 8 loci being doubtful, 29 antigens being incorrectly interpreted by serology and 20 of serological blanks turning out to be definable alleles by the DNA method. Conclustons Genotyping for HLA-DR by PCR-SSP offers the advantages of better reagent and sample availability, more rapid and greater accuracy, all of which would warrant that this approach was suitable for clinical Practice in organ transplantation.