自制HLA—B27基因检测试剂盒的应用

Application of the Self-made Human Leukocyte Antigen HLA-B27 Test Kits

目的检验自制HLA-B27基因检测试剂盒的可靠性。方法根据序列特异性引物聚合酶链反应（SSP—PCR）原理，通过对HLA数据库上HLA—B位点基因序列的比对，分析选择出B27特异序列合成引物，进而对PCR反应体系和条件进行摸索优化，自制HLA-B27基因检测试剂盒，同时以北京思而成B27基因检测试剂盒为对照试剂盒，对254例疑似强直性脊椎炎、急性葡萄膜炎等患者EDTA抗凝血的B27基因进行检测，结果和对照试剂盒进行比较，分析其一致性。结果自制试剂盒疑似患者的HLA-B27阳性检出率为35．8％，对照试剂盒的检出率36．6％，经配对x^2检验，x^2=0．125，P=0．727〉0．05，两试剂盒阳性检出率差异无统计学意义。结论自制试剂盒的成本低，具有一定的实验室研究应用价值。

Objective To verify the accuracy of the self-made human leukocyte antigen B27 （HLA-B27） detect kit. Methods According to the principle of sequence specific primer polymerase chain reaction （SSP-PCR） ,matched the HLA sequence on database of HLA, designed the B27 primer sequences to specific amplification of B27 fragment, and thus to optimize the PCR conditions and reaction system,self-made B27 test kit to detect B27 gene from EDTA anticoagulant blood of 254 cases of suspected ankylosing spondylitis, acute uveitis, the test results was compared with control kits, and analysed the consistency of them. Results No significant differences were found between two kind kits（x2 =0. 125,P=0. 727〉0.05）. Conclusion A new simple, fast and low cost kit is recommended in clinical application in the future.