Establishment of Q-PCR method for qualitative testing of the presence or absence of KIR genes
10.13303/j.cjbt.issn.1004-549x.2024.06.009
- VernacularTitle:应用Q-PCR定性检测KIR基因有无方法的建立
- Author:
Yunan LI
1
;
Jianxin ZHEN
;
Shuang LIANG
;
Qiong YU
;
Zhihui DENG
Author Information
1. 深圳市血液中心 输血医学研究所,广东 深圳 518040
- Keywords:
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)
- From:
Chinese Journal of Blood Transfusion
2024;37(6):660-665
- CountryChina
- Language:Chinese
-
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.
