Concordance of identifying the presence or absence of KIR genes by Flow-rSSO and PCR-SBT methods: a comparative study
10.13303/j.cjbt.issn.1004-549x.2023.07.002
- VernacularTitle:Flow-rSSO及PCR-SBT方法检测KIR基因有无的对比研究
- Author:
Fuling ZHONG
1
;
Zhichao YANG
1
;
Hao CHEN
1
;
Zhihui DENG
1
Author Information
1. Shenzhen Blood Center, Shenzhen 518035, China
- Publication Type:Journal Article
- Keywords:
KIR genes;
sequencing-based typing-PCR (PCR-SBT);
flow reverse sequence specific oligonucleotide probe (Flow-rSSO);
sequence specific primers-PCR (PCR-SSP)
- From:
Chinese Journal of Blood Transfusion
2023;36(7):563-566
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To study the concordance of identifying the presence or absence of KIR genes using flow reverse sequence-specific oligonucleotide probe (Flow-rSSO) hybridization and sequencing based typing-PCR (PCR-SBT) methods. 【Methods】 A total number of 131 cases of DNA samples from Han population were subjected to identify the presence or absence of all 16 KIR genes by Flow-rSSO method, and then sequenced at coding sequence for all 14 functional KIR genes using our in-house KIR PCR-SBT assay. The concordance of identifying the presence or absence of all functional KIR genes by Flow-rSSO and PCR-SBT was analyzed. Samples with inconsistent initial results were re-tested using the Flow-rSSO commercial kits with different Lot number, and further tested using the PCR-SSP commercial kit. 【Results】 The presence or absence of 14 functional KIR genes for 129 of 131 samples were completely in accordance via the PCR-SBT and Flow-rSSO methods. Two samples, one with 3DL1 negative, the other with both 2DS3 and 2DS5 negative initially-identified by Flow-rSSO, were actually all positive tested by PCR-SBT. Further retest by Flow-rSSO commercial kits with different Lot number and PCR-SSP commercial kit indicated that the two samples were all positive, which agreed well with PCR-SBT results. 【Conclusion】 In this paper, the initial test results of the presence or absence of KIR genes identified by Flow-rSSO for 2 samples were wrong, which indicated the importance of carrying out the quality control for reagents in KIR gene testing.
