Application of a simplified PCR-SSP method to detect A4GALT*01 and A4GALT*02 typing among Thai blood donors.

OBJECTIVES

An intronic A4GALT single nucleotide variant, rs5751348:G>T, P or A4GALT02 allele has a lower level of the enzyme-encoding A4GALT transcripts than the P individuals. Here, we first develop and validate a simple inhouse PCR-SSP method to detect A4GALT01 and A4GALT*02 alleles, and second, apply this method to compare the allele frequencies between Thai and other populations.

MATERIAL AND METHODS

The conventional test tube technique was used to detect the P1 antigen in 222 blood samples from Thai blood donors at Thammasat University Hospital. A PCR-SSP method was optimized and validated for reproducibility and specificity to identify these alleles and was subsequently tested on 1,840 DNA samples of unknown phenotypes obtained from central, northern and southern Thais. In addition, allele frequencies of central Thais were compared with those of other populations.

RESULTS

In the tested cohort (n = 222), P and P phenotypes were typed in 26.13 and 73.87% of donors, respectively. The developed PCR-SSP was successfully optimized, and the outcomes were consistent with those of serological phenotyping and DNA sequencing results, demonstrating its validity for predicting P/P phenotype. For central, northern and southern Thais, the A4GALT01 frequency was 0.1579 (430/2,724), 0.1183 (71/600), and 0.2575 (206/800), whereas the A4GALT02 frequency was 0.8421 (2,294/2,724), 0.8817 (529/600), and 0.7425 (594/800), respectively. Their observed frequencies among central Thais significantly differed from those in other populations (p < 0.05).

CONCLUSION

Our study has successfully developed a simple, precise, and reliable method to genotype A4GALT01 and A4GALT02 using inhouse developed PCR-SSP for predicting P/P status.