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DNA profiling of microdissected spermatozoa |
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| รหัสดีโอไอ | |
| Creator | Adcharee Kongruang, Jittima Shotivaranon, Prapatsorn Areesirisuk, Anna Wongkularb, Budsaba Rerkamnuaychoke |
| Title | DNA profiling of microdissected spermatozoa |
| Contributor | Adcharee Kongruang, Jittima Shotivaranon, Prapatsorn Areesirisuk, Anna Wongkularb, Budsaba Rerkamnuaychoke |
| Publisher | Genetics Society of Thailand |
| Publication Year | 2564 |
| Journal Title | Genomics and Genetics |
| Journal Vol. | 14 |
| Journal No. | 1 |
| Page no. | 9-17 |
| Keyword | DNA profile, laser microdissection, spermatozoa, sexual assault |
| URL Website | https://li01.tci-thaijo.org/index.php/gst/issue/view/17009 |
| Website title | https://li01.tci-thaijo.org/index.php/gst/article/view/243350 |
| ISSN | 24655198 |
| Abstract | In sexual assaultcases, DNA profiling for perpetrator identification requires isolation of pure spermatozoa because evidences are often mixture between the sperm cells and the victim's cells. Laser microdissection is a powerful tool that directly separates and collects the sperm cells without contaminationfromanother cell. The objectivesof this report areto study DNA profiling of laser microdissected spermatozoa for personal identification and to study different factors that may affect the interpretation of DNA profilesfrom laser microdissected spermatozoa. First, we demonstratedthe ability of laser microdissection to identify and genotype spermatozoa. Second, three staining reagents; modified Hematoxylin & Eosin (H&E), eosin and sperm HY-LITERฎ, were evaluated on quality of DNA profilesthat derived from laser microdissected spermatozoa. Finally, STR analysis was performed on laser microdissected cell from stained slides that stored in various time.The results showed that at least 150 sperm cells were required to obtain a complete DNAprofile, whereas, a partial DNA profile was obtained from 25-75 haploid sperm cells. In staining comparisonresult, modified H&E providedthe best result of DNA profilesand suitableto apply forlaser microdissection. In addition, we obtained complete DNA profilesfrom stained slidesstored for 1 month whereas the stained slidesstored for 12 monthsyielded poor DNA profiles. It showed that storagetime of the stained slides hadan effect to DNA quality and DNA analysis.In conclusion, laser microdissection is a useful tool to isolate sperm cells and makes a chance to generate genetic profile from low number of sperm cells. However, number of starting sperm cells, staining and duration of storage time are factors that affect the quality of the DNA profile derived from laser microdissected spermatozoa. In addition,to obtain reliable DNA profiles, it is recommended toavoid the factors that may havenegative effect on DNA typing process. |
