Development of a Multiplex PCR for Human Neutrophil Antigens-1, -3, -4, and -5 Genotyping Among Thai Blood Donors
Autor: | Wiradee Sasikarn, Kamphon Intharanut, Siriporn Nathalang, Kanokpol Siriphanthong, Oytip Nathalang |
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Rok vydání: | 2017 |
Předmět: |
Isoantigens
Time Factors Genotype Neutrophils Cost-Benefit Analysis Blood Donors 030204 cardiovascular system & hematology Biology Lung injury Neutropenia General Biochemistry Genetics and Molecular Biology DNA sequencing law.invention Workflow 03 medical and health sciences 0302 clinical medicine law Multiplex polymerase chain reaction medicine Humans Genotyping Polymerase chain reaction Human Neutrophil Antigens Reproducibility of Results Health Care Costs medicine.disease Thailand Virology Molecular biology Healthy Volunteers 030220 oncology & carcinogenesis Multiplex Polymerase Chain Reaction |
Zdroj: | Clinical laboratory. 62(11) |
ISSN: | 1433-6510 |
Popis: | Background Different polymerase chain reaction (PCR) techniques for human neutrophil antigens (HNA) genotyping have been implemented to diagnose the clinical conditions of patients with alloimmune neutropenia, febrile non-hemolytic transfusion reactions, and transfusion-related acute lung injury and to provide effective HNAmatched granulocyte transfusions. The present study aimed to develop an in-house multiplex-PCR for HNA-1, -3, -4, and -5 genotyping in the Thai population. Methods Altogether, 500 DNA samples obtained from unrelated, healthy Thai blood donors at the National Blood Centre, Thai Red Cross Society, Bangkok, Thailand were included. Three hundred DNA samples of known HNA genotyping based on PCR with sequence specific primers (PCR-SSP), as previously described, were tested with the newly developed multiplex PCR. Additionally, 200 DNA samples of unknown HNA genotyped donors were tested for HNA-1, -3, -4, and -5 genotyping using multiplex-PCR. Results Validity of HNA-1, -3, -4, and -5 genotyping by multiplex PCR using known DNA controls and the comparison of the genotyping results between PCR-SSP and multiplex PCR were in agreement. Interestingly, the rare genotype HNA-4b4b was not found in this study, similar to previous studies in Thai and other populations. Moreover, 30 samples were randomly tested twice for HNA genotyping using the multiplex-PCR and demonstrated reproducible results and were confirmed by DNA sequencing. Conclusions This study shows that the newly developed multiplex-PCR is cost effective and less time consuming compared with PCR-SSP. The multiplex PCR can be used as an alternative technique for HNA-1, -3, -4, and -5 genotyping for routine testing, especially in other developing countries due to its simplicity and accuracy. |
Databáze: | OpenAIRE |
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