中国输血杂志2024，Vol.37Issue(6)：660-665,6.DOI:10.13303/j.cjbt.issn.1004-549x.2024.06.009
应用Q-PCR定性检测KIR基因有无方法的建立
Establishment of Q-PCR method for qualitative testing of the presence or absence of KIR genes
摘要
Abstract
Objective To establish a method for qualitative detection of the presence or absence of all KIR genes by quantitative polymerase chain reaction(Q-PCR).Methods Based on the polymorphism of high-resolution level KIR alleles in Chinese population and the IPD-KIR database,KIR gene-specific primers were designed to amplify all the 16 KIR genes and 2DS4-Normal and 2DS4-Deleted subtypes by Q-PCR.Meanwhile,one negative control and one positive control specific amplifying human growth hormone(HGH)gene fragment were set to monitor the false positive and false negative results in PCR amplification,respectively.A total of 302 samples with known KIR genotype previously identified by KIR PCR-SSP commercial kit were randomly selected for blind inspection to verify the reliability of KIR Q-PCR method established by au-thors.Results The results of 300 samples detected by our KIR Q-PCR method were consistent with the known results,but two samples showed inconsistent results.One sample was negative for 2DS5 by Q-PCR but positive by PCR-SSP,another sample was positive for 2DS1 by Q-PCR but negative by PCR-SSP.The two doubtful samples were genotyped by sequencing-based typing(PCR-SBT)for 2DS5and2DS1,respectively.PCR-SBT results confirmed that the results of Q-PCR test was correct.Conclusion The KIR Q-PCR method established in this paper can provide accurate and reliable results for testing the presence or absence of KIR genes.关键词
杀伤细胞免疫球蛋白样受体(KIR)/KIR基因有无/实时荧光定量-PCR/序列特异性引物-PCR/测序分型Key words
killer-cell immunoglobulin-like receptor(KIR)/presence or absence of KIR genes/quantitative real time-PCR(Q-PCR)/sequence specific primers-PCR(PCR-SSP)/sequencing-based typing(PCR-SBT)分类
医药卫生引用本文复制引用
李宇楠,甄建新,梁爽,喻琼,邓志辉..应用Q-PCR定性检测KIR基因有无方法的建立[J].中国输血杂志,2024,37(6):660-665,6.基金项目
国家自然科学基金青年科学基金项目(82203291) （82203291）
广东省基础与应用基础研究基金委员会项目(2022A1515011045) （2022A1515011045）
深圳市科技创新委员会基础研究项目(JCYJ20190806152001762) （JCYJ20190806152001762）
深圳市医学重点学科(SZXK070) （SZXK070）
