Open AccessA New Method for Analyzing the Duffy Blood Group Genotype by TaqMan Minor Groove Binding Probes
Author(s) -
Zhou Shihang,
Liu Ming,
An Wanxin,
Liang Xiaohua,
Yu Weijian,
Piao Fengyuan
Publication year - 2015
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.21751
Subject(s) - genotyping , taqman , genotype , microbiology and biotechnology , polymerase chain reaction , concordance , biology , allele , locus (genetics) , snp genotyping , minor groove , population , genetics , dna , medicine , gene , environmental health
Background Duffy blood group genotyping is useful to ensure transfusion safety and determine the association of Duffy blood group polymorphism with diseases, and therefore has its clinical significance. In order to improve the existing methods for genotyping of Duffy blood group, which normally require post‐PCR manipulation, a new method was developed by using 5′‐nuclease assay (NA) with TaqMan minor groove binding (MGB) probes. Methods Primers and TaqMan‐MGB probes were designed and synthesized to genotype FY*A and FY*B alleles at Duffy blood group locus on a real‐time PCR platform. A total of 120 samples were genotyped by using the new 5′‐NA and conventional polymerase chain reaction with allele‐specific primers (PCR‐ASP). The results obtained by the two methods were compared. Results There was a complete concordance of results for all samples genotyped by 5′‐NA and PCR‐ASP. The retesting results of 5′‐NA were consistent with those of the initial testing. The detection limit of 5′‐NA was determined as 100 pg per reaction. The FY*A and FY*B allelic frequencies were 93.3% and 6.7% respectively in the Chinese Han population in Dalian. Conclusions The 5′‐NA for genotyping of Duffy blood group is simple, rapid, reliable, reproducible, sensitive, and high‐throughput and is superior to PCR‐ASP used in routine genotyping.