These authors contributed equally to this work.
Multiplex polymerase chain reaction with DNA pooling: a cost-effective strategy of genotyping rare blood types
Article first published online: 29 OCT 2012
© 2012 The Authors. Transfusion Medicine © 2012 British Blood Transfusion Society
Volume 23, Issue 1, pages 42–47, February 2013
How to Cite
He, Y.-L., Gao, H.-H., Ye, L.-Y., Guo, Z.-H., Wang, P. and Zhu, Z.-Y. (2013), Multiplex polymerase chain reaction with DNA pooling: a cost-effective strategy of genotyping rare blood types. Transfusion Medicine, 23: 42–47. doi: 10.1111/j.1365-3148.2012.01198.x
- Issue published online: 22 JAN 2013
- Article first published online: 29 OCT 2012
- Manuscript Accepted: 20 SEP 2012
- Manuscript Revised: 9 JUL 2012
- Manuscript Received: 20 APR 2012
- International Cooperation Program of Science and Technology Commission of Shanghai Municipality, China. Grant Number: 08410702500
- Research Foundation of Shanghai Health Bureau, China. Grant Number: 2009194
- Health Care, China. Grant Number: 201002005
- blood typing;
- DNA pool;
This work aims to develop a multiplex polymerase chain reaction combined with DNA pooling for mass screening for rare blood types.
The differences in most blood group antigens are associated with single-nucleotide polymorphisms (SNPs), which are used in detecting blood antigen expression at the molecular level. However, all existing sequence-specific primers polymerase chain reaction (PCR-SSP) assays for blood typing genotype one or several SNPs individually. DNA pooling is a way that reduces the amount of genotyping required.
A sensitive multiplex PCR-SSP assay testing pooled DNA was established to detect the rare Fyb and S alleles. It was applied to screen a total of 4490 donor samples via testing 898 DNA pools. The samples in the positive pools were further tested individually. Then the positive samples, including Fy(a−b+)/Fy(a+b+) and S+s−/S+s+ genotypes, were tested via two PCR-SSP assays for alleles Fya and s. The rare genotypes Fy(a−b+) and S+s− were verified using serologic tests and sequencing analysis.
Two hundred and fifty-four donors were tested positive for the Fyb allele, whereas 101 donors were positive for the S allele. Among the 254 Fy(b+) donors, 5 were Fy(a−b+) and 249 were Fy(a+b+). Among the 101 S+ donors, 3 were S+s− and 98 were S+s+. The rare Fyb and S alleles comprised 2·28 and 1·16%, respectively. The PCR-SSP assays were confirmed by sequencing analysis and serological test.
A multiplex PCR assay was combined with DNA pooling to reduce the number of tests required, making large-scale screening feasible.