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Published on: September 11, 2022
[Detection for KIR gene with use of SYBR green I real-time PCR]
Lian-hong Xu1, Yong-fang Wang, Chuan-ping Qi
1Department of Clinical Laboratory, Affiliated Wujin People's Hospital of Jiangsu University, Changzhou 213002, China. lotus2104@yahoo.com.cn
Aim:
To develope a novel real-time PCR method for KIR genotyping.
Methods:
KIR genotyping is performed using 16 real-time PCR reactions, each containing two-three KIR-specific primers, a pair of internal positive control primers and a fluorescence dye, SYBR Green I. By the analysis of the Tm and the characteristic of the melting curve of the amplified products, we identified the presence or absence of 16 KIR genes. A variety of dilution folds were made to detect the sensitivity of this method.
Results:
KIR genes were effectively genotyped by the analysis of the melting curve. This method can be used to detect KIR genes even from 0.1 ng of DNA. The feasibility of this method was tested by genotyping 10 DNA samples from Peripheral blood and 10 DNA samples from cervical cell.
Conclusion:
We developed a novel real-time PCR assay with SYBR Green I, which is a simple, rapid, sensitive, real-time and environmental method. It provides the possibility of the automated KIR genotyping.
