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目的建立检测ＨＬＡ－Ｂ２７等位基因的聚合酶链反应／顺序特异的寡核苷酸探针方法，并分析其实用性。方法采用４对引物Ｅ４０ｓ、Ｅ９０ａｓ；Ｅ９１ａｓ、Ｅ１３６ａｓ；Ｅ９１ｂｓ、Ｅ１８１ａｓ；Ｅ４０ｓ、Ｅ１３０ａｓ分别扩增靶ＤＮＡ，用１０个探针ＣＬ－１～９和ＰＡＮ－Ｂ２７分别进行杂交，探针用地高辛系统标记和检测，可检定Ｂ＊２７０１～２７０８共８个等位基因。结果通过对Ｂ２７等位基因已明确的８个阳性对照ＤＮＡ、５例Ｂ２７阴性标本及７５例阳性标本的检测，表明本方法技术稳定结果可靠，不会与其它ＨＬＡⅠ类基因产生交叉杂交而影响结果的指定。结论本研究为ＨＬＡ－Ｂ２７多态性和强直性脊椎炎关联研究提供了一种有效方法，也为ＨＬＡⅠ类分子基因分型研究积累了经验。
Objective To establish a polymerase chain reaction / sequence specific oligonucleotide probe for detection of HLA-B27 alleles and to analyze its practicability. Methods Four pairs of primers E40s, E90as, E91as, E136as, E91bs, E181as, E40s and E130as were used to amplify the target DNA. Hybridization was carried out with 10 probes CL-1-9 and PAN-B27 respectively. Marking and detection, can be detected B * 2701 ~ 2708 a total of eight alleles. Results Eight positive control DNA, five B27 negative samples and 75 positive samples were identified by the B27 allele. The results showed that the method was reliable in technique stability and could not affect the results by cross-hybridization with other HLA class I genes The designation. Conclusion This study provides an effective method for the association between HLA-B27 polymorphism and ankylosing spondylitis, and also accumulates experience in the genotyping of HLA class I molecules.