| A Study of HLA-DQA Genotyping of Hair DNA Using the PCR Method |
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Jae Hong You1, Keon Su Lee1, Jong Woo Park2 |
1Department of Pediatrics, Chungnam National University College of Medicine Taejon, Korea
2Department of Clinicopathology, Chungnam National University College of Medicine Taejon, Korea |
| 모발에서의 중합효소 연쇄반응을 이용한 HLA-DQA 유전자형 판별에 관한 연구 |
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유재홍1, 이건수1, 박종우2 |
1충남대학교 의과대학 소아과학교실
2충남대학교 의과대학 임상병리학교실 |
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| Abstract |
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The characterization of genetic variation at the level of DNA has generated significant advances in gene mapping and disease diagnosis, and forensic identification of individuals. It is now possible to identify individual DNA from various tissue specimens, like hair, using the PCR and oligonucleotide probes.
To date, however, the number of hairs needed, the preservation conditions, and the kinds of hair suitable for DNA extraction have not been well known. We performed DNA extraction using hairs from different body sites, using different numbers of hairs, under various different preservation conditions to investigate the acquisition conditions of DNA data from hair using PCR and specific HLA-DQA probe. HLA-DQA genotyping of DNA extracted from peripheral blood was performed to compare the results of hair and blood HLA-DQA genotyping from individuals.
The results are as follow:
1) The concentration of DNA extracted from a single strand of hair is 5.23±0.54ug/ml. It is possible to extract sufficient DNA for HLA-DQA genotyping from a single strand of hair.
2) DNA concentration is different according to body site. Concentrations are 7.01±0.33ug/ml in scalp hair, 6.28±0.29ug/ml in axillary hair, and 6.10±0.24ug/ml in pubic hair.
3) There is no difference between the electrophortic bands resulting from DNA extracted from the hair of an individual preserved under different conditions, such as room temperature, exposure to sunlight, exposure to low temperature (+40℃), or exposure to moisture.
4) There is no difference between the electrophoretic bands resulting from DNA extracted from hair of a single individual preserved for different lengths of time.
5) In an individual, the HLA-DQA genotype obtained from peripheral blood is identical to that obtained from hair.
Even though the amout of DNA obtained from hair is limited, it is possible to identify the HLA-DQA genotype of an individual using a single strand of hair. This requires adequate extraction of DNA ofr PCR analysis using an allele specific probe. We believe that HLA-DQA genotyping using the PCR method on DNA extracted from hair is useful for disease diagnosis and forensic science. |
| Key Words:
HLA-DQA genotyping, Hair, PCR |
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