用PCR-SSP分析HLA-B位点血清学分型困难标本

The analysis of ambiguous sera-typing for HLA-B locus with PCR-SSP

目的 :对血清学HLA B位点分型困难标本用PCR SSP方法分析 ,并寻找血清分型效果不佳原因。方法 :用本室建立的PCR SSP方法对 10 0株标准细胞及 91份血清学分型困难标本分别作HLA B位点分析 ,前者作为PCR SSP方法的参考对照品。结果 :10 0株标准细胞分析结果与已知结果一致 ;91份疑问标本均顺利地用PCR SSP分型 ,结果共有 70种等位基因格局 ,其中 36种与血清学完全不同 ,占 5 2 %。PCR SSP还检出了一些原先认为上海人中空白的基因 ,如B49。结论 :从PCR SSP分型结果推断血清学部分标本分型结果不佳的原因是缺少单价抗血清 ;同一交叉反应组内错检 ;反应强度不当 ,导致结果难辨 ;操作中跨孔污染。而PCR SSP就没有这方面的问题。证实了本室建立的PCR SSP法可作为准确可靠的HLA

Objective:To analyse HLAB locus of 91 ambiguous seratyping samples with PCRSSP method and find out what cause seratyping ambiguous.Methods:HLAB locus of 91 ambiguous seratyping samples and 100 reference cell lines as reference were analysed with PCRSSP method which was established in the lab.Results:91 samples were successfully typed with PCRSSP,totally 70 different result patterns were gotten,among which 36 patterns were different from those of seratyping,counting for 52%.Besides,PCRSSP can detect some alleles which were formerly thought as blank in shanghaiese ,for example B49.Conclusion:The reasons for the differences were deduced as:no monospecific antisera was available.Mistyping among cross reactive group antigens.The too strong or weak reactions between antigens and antisera interfered with the determination of results.The cross over within different wells in operating.PCRSSP is free from these problems listed above and the accuracy of PCRSSP was demonstrated by the completely result coincidence with reference cell lines.PCRSSP is confirmed as a more accurate and reliable method for HLAB typing,satisfying the clinical needs.