Red Cell Genotyping by Multiplex PCR Identifies Antigen-Matched Blood Units for Transfusion-Dependent Thai Patients

Abstract

Background: Antigen-negative red cell transfusion is required for transfusion-dependent patients. We developed multiplex PCR for red cell genotyping and calculated the possibility of finding compatible predicted phenotypes in Thai blood donor populations according to red cell alloantibodies found among Thai patients. Methods: 600 DNA samples obtained from unrelated healthy central and northern Thai blood donors were tested with the newly developed multiplex PCR for FY*A, FY*B, JK*A, JK*B, RHCE*e, RHCE*E, DI*A and GYP*Hut, GYP*Mur, GYP*Hop, GYP*Bun, and GYP*HF allele detections. Additionally, the possibility of finding compatible predicted phenotypes in two Thai blood donor populations was calculated to estimate the minimal number of tests needed to provide compatible blood. Results: The validity of multiplex PCR using known DNA controls and the phenotyping and genotyping results obtained by serological and PCR-SSP techniques were in agreement. The possibility of finding at least one compatible blood unit for patients with multiple antibodies was comparable in Thai populations. Conclusions: The multiplex PCR for red cell genotyping simultaneously interprets 7 alleles and 1 hybrid GP group. Similar strategies can be applied in other populations depending on alloantibody frequencies in transfusion-dependent patients, especially in a country with limited resources.

Determining of JK*A and JK*B Allele Frequency Distribution among Muslim Blood Donors from Southern Thailand

“…The results of a two-tube PCR-SSP were used to distinguish between JK*A and JK*B alleles. The first and second mixes could differentiate between JK*A and JK*B alleles with an amplified product size of 301 bp, similar to the results of a related study ( 15 ). The validated genotyping results of 10 DNA controls were consistent with each other, and 20 DNA samples tested by PCR-SSP showed 100% concordance with the DNA sequencing results.…”

“…Detection of JK*A and JK*B alleles was carried out using standard PCRSSP, as previously described, with some modifications ( 15 ). In brief, 1 μL of genomic DNA (50 ng/μL) was amplified in 10 μL of total volume (1 μL of 5 μM JK-AB-Forward primer 5′-CATGCTGCCATAGGATCATTGC-3′ and 1 μL of 5 μM JK-A-Reverse primer 5′-CCAGAGTCCAAAGTAGATGTC-3′) to detect the JK*A allele.…”

Genotyping for Dombrock blood group alleles in Northern Pakistani blood donors

“…Compared with classic serologic testing methods, molecular methods have several advantages. 12 Serologic procedures are costly, and various antisera are not available commercially, 21 whereas DNA-based methods are quicker and attain a higher throughput at a lower cost. 22 Genotyping can aid in donor screening for both common and rare blood group antigens, 8 thereby increasing the probability of providing antigen-specific blood products to patients.…”

“…To perform a molecular study, we had to consider several factors such as finances; time taken per test; throughput, sensitivity, and specificity of the tests; and equipment availability. Several high-throughput DNA platforms are marketed-for example, Blood Chip, HEA Bead Chip, Luminex XMAP, and so forth-but they are very costly 21 and are thus inappropriate for a country with insufficient resources like Pakistan. Therefore, we chose PCR-SSP for our study.…”

Hybrid glycophorin and red blood cell antigen genotyping in Asian American type O blood donors with Mia phenotype

“…The multiplex ligation‐dependent probe amplification method was used in donor screening in Guangzhou, China . The multiplex SSP‐PCR method was developed for genotyping five blood groups, including MNS hybrid glycophorins in two Thai populations . The real‐time PCR melting curve method has been used to identify the GYP*Mur allele from other Mi a ‐positive phenotypes .…”