Tan J, Xie T, Xu Q
Department of Urology, Shanghai First People's Hospital.
Zhonghua Yi Xue Za Zhi. 1997 Jan;77(1):28-30.
DNA typing for HLA-DR by polymerase chain reaction with sequence-specific primers (PCR-SSP) compared with standard serology to evaluate the reliability and clinical practicability.
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 cadaveric transplantation. Matching time, sensitivity, specificity and clinical practicability were compared according to typing results by both methods.
All 162 samples were able to be typed by PCR-SSP. A total of 308 alleles were detected (16 DR "blank"). The results of matching were confirmed by analysis with restriction endonucleases and Southern 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 30.2% (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.
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.
采用序列特异性引物聚合酶链反应(PCR-SSP)对HLA-DR进行DNA分型,并与标准血清学方法进行比较,以评估其可靠性和临床实用性。
对61例尸体供者和101例尸体移植受者,采用PCR-SSP法进行DNA分型及采用微量淋巴细胞毒技术进行标准血清学分型,进行HLA-DR等位基因的双盲分型。根据两种方法的分型结果,比较配型时间、敏感性、特异性及临床实用性。
162份样本均能用PCR-SSP法进行分型。共检测到308个等位基因(16个DR“空白”)。酶切分析及Southern杂交证实了配型结果。特异性和重复性均为100%。PCR-SSP法进行HLA-DR分型需5小时,血清学法需20小时。PCR-SSP法与血清学HLA-DR分型的差异率为30.2%(肾移植受者为35.6%,供者为21.3%)。差异包括8个位点可疑、29个抗原血清学误判及20个血清学“空白”经DNA方法检测为可确定的等位基因。
PCR-SSP法进行HLA-DR基因分型具有试剂和样本易获取、速度更快、准确性更高的优点,所有这些都表明该方法适用于器官移植的临床实践。