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Human DNA identification and sex determination from bloodstains by duplex PCR analysis |
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| รหัสดีโอไอ | |
| Creator | Khemika Lomthaisong |
| Title | Human DNA identification and sex determination from bloodstains by duplex PCR analysis |
| Contributor | Potchaninporn Amornpan, Apichaya Petthai, Monthira Monthatong |
| Publisher | Research and Technology Transfer Affairs Division.Khon Kaen University. |
| Publication Year | 2560 |
| Journal Title | Asia-Pacific Journal of Science and Technology (APST) |
| Journal Vol. | 22 |
| Journal No. | 4 |
| Page no. | 1-10 |
| Keyword | Duplex PCR, Bloodstains, Cytochrome b, SRY |
| URL Website | https://tci-thaijo.org/index.php/APST/index |
| Website title | https://tci-thaijo.org/index.php/APST/article/view/107639 |
| ISSN | 2539-6293 |
| Abstract | This study aimed to identify DNA of human origin and determine human sex from bloodstains by duplex PCR analysis. Two pairs of primers amplifying Amelogeningene, AMEL(1) and AMEL(2) and a primer pair targeting Sex Determining Region Ygene [SRY] were compared at the preliminary stage to measure the accuracy of sex determination. Human specific region of Cytochrome b(Cyt b) gene sized 412 bpsuccessfully targeted only human-origin DNA, whileother animal DNAs (dog, cat, cow, chicken and pig) did not produce any PCR product. SRY primer showed 100% accuracy on male identification with 197 bp amplicon. Therefore, the SRY primer pair was chosen for sex identification. The optimal annealing temperature of duplex PCR analysis was 55?C, producing the most distinctive PCR products, determining male with 197 bp and 412 bp bands and female with only 412 bp. For sensitivity, the smallest blood sample that could be detected was 20 ?l. For specificity, both human origin and sex determinationcould be detected in all ratios of mixed human and dog bloods, including using 100 fold less human blood to animal blood. Determination of male blood in mixed male and female blood samples was also investigated. The results showed that male blood could bedetermined in mixed female blood sample at ratios less than 10 fold. This study demonstrated that duplex PCR analysis of Cyt band SRYis a reliable tool to investigate human DNA and sex of questioned bloodstains. |
