人體各類骨骸DNA檢出情形之研究

依文獻資料顯示，人體各種不同部位骨骼DNA量及DNA型別檢出率之結果有明顯差異。影響骨骼DNA鑑定結果的因素包括骨骼腐敗程度、骨骼的種類、死者年齡、DNA萃取法、PCR抑制物、檢體是否污染及DNA鑑定試劑套組等。依實務經驗認為未能成功檢出骨骼DNA型別的主要原因之一，乃是採集骨骼的DNA含量太少。此外，採用何種DNA萃取法亦是影響因素之一。本研究以改良式Amicon萃取法評估各類骨骸DNA含量及DNA型別檢出率。採集99件無名屍骨骸（包含8組頭顱骨計48件），依形態分為長骨、不規則骨及扁平骨（腦顱骨），以Quantifiler® Human Kit進行DNA定量，分析各類骨骼DNA量、STR及Y-STR DNA型別，比較各類骨骼之DNA型別檢出率。研究結果說明如下：一、DNA型別檢出率以長骨最高（92～100%），不規則骨次之（88～94%），扁平骨最低（55～83%）；二、分析頭顱的上頷骨、下頷骨（面顱骨）及扁平骨（腦顱骨）之STR DNA檢出率分別為93%、88%及68%；三、分析各種腦顱骨之STR DNA型別檢出率由高至低依序為顳骨、枕骨、頂骨及額骨，分別為83%、69%、64%及55%。綜合各種骨骼DNA量、DNA型別檢出率、研究案例及本實驗室先前法醫牙齒之DNA鑑定研究結果，如為一具完整骨骸，建議優先採集長骨及牙齒，不規則骨次之，頭顱骨再次之。如僅存頭顱1顆，建議採檢牙齒及上、下頷骨（面顱骨）。而腦顱骨以顳骨較佳。研究成果可提供第一線法醫人員採集骨骼檢體優先順序之參考，避免重新採樣，以提升鑑定效率。

關鍵字

骨骼；頭顱； Amicon萃取法； DNA定量；牙齒

並列摘要

The research shows that there are evident differences of DNA quantity and DNA profile detection rate for various parts of skeletons. The factors which have effect on DNA identification of skeleton include degree of decomposition, categories of skeleton, age-of-death, DNA extraction method, PCR inhibitors, contamination, and also used DNA identification kits. Based on our practical experience, the failure at DNA profile detection in human skeleton is not only mainly caused by deficit DNA quantity in selected bones but also the applied DNA extraction method. Our study used the modified Amicon extraction method, Quantifiler® Human Kit, STR and Y-STR DNA profiles to evaluate DNA quantity and DNA profile detection rates on 99 different parts of skeletons from unidentified bodies cases. The results show that (1) among different kinds of bones, the highest DNA profile detection rates was found in long bone (92-100%), followed by irregular bone (88-94%), and the lowest was flat bone (55-83%); (2) for the skull, DNA profile detection rates of maxilla, mandible (facial bone), and cranial skeletons (flat bone) were 93%, 88%, and 68%, respectively; (3) the DNA profile detection rates for cranial skeletons were analyzed and found decreased in following order: temporal (83%), occipital (69%), frontal (64%), and parietal (55%) bone. To sum up the results of DNA quantity and DNA profile detection rats in various categories of skeleton, practical experience, along with our previous research about DNA identification for teeth, the long bones and teeth are suggested to be given priority, followed by irregular bones and skulls as full body skeleton are available. If only skull is able to obtained, the teeth, maxilla, and mandible bone should be considered first. When only cranium are founded, the temporal bone should be the first choice. This study provids an order of precedence among different human skeletal remains for sampling so as to avoid time-wasting repeated sampling and improve the efficiency of forensic DNA identification.

並列關鍵字

Skeleton ； Skull ； Amicon Extraction ； DNA Quantification ； Teeth

參考文獻

黃純英、蔡華生、鍾芳君等：以Amicon濃縮管萃取骨骸DNA之研究。台灣法醫學誌2018;10:4-14. doi:10.6134/tjfm.201812_10(2).0002

Amory S, Huel R, Bilić A, Loreille O, Parsons TJ: Automatable full demineralization DNA extraction procedure from degraded skeletal remains. Forensic Sci Int Genet. 2012;6:398-406. doi:10.1016/j.fsigen.2011.08.004

Kuś M, Ossowski A, Zielińska G: Comparison of three different DNA extraction methods from a highly degraded biological material. J Forensic Leg Med 2016;40:47-53. doi:10.1016/j.jflm.2016.03.002

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人體各類骨骸DNA檢出情形之研究

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