Aly Sanaa Mohamed, Wen Jifang, Wang Xiang
Department of Forensic Science and Pathology, School of Basic Medical Sciences, Central South University, Changsha 410013, Hunan, China.
Am J Forensic Med Pathol. 2013 Jun;34(2):159-63. doi: 10.1097/PAF.0b013e31828c390e.
Entomological evidence is of great importance in forensic cases for postmortem interval calculation. The use of Sarcophagidae (Diptera) for postmortem interval estimation is limited because morphological determination is often hampered because of similar characteristics in the larval, pupal, and even adult stage. To make the species identification more accurate and reliable, DNA-based identification is considered. In this study, we assessed the use of partial mitochondrial cytochrome oxidase I and II genes for discrimination of forensically important Sarcophagidae from Egypt and China [Sarcophaga argyrostoma (Robineau-Desvoidy), Sarcophaga dux (Thomson), Sarcophaga albiceps (Meigen), and Wohlfahrtia nuba (Wiedemann)]. This region was amplified using polymerase chain reaction followed by direct sequencing of the amplification products and using restriction enzymes HinfI and MfeI. Nucleotide sequence divergences were calculated using the Kimura 2-parameter distance model, and a neighbor-joining phylogenetic tree was generated. All examined specimens were assigned to the correct species. Combinations of the restriction enzymes HinfI and MfeI provide different restriction fragment length polymorphism profiles even among 3 sympatric species that belong to the Sarcophaga genus. Therefore, this study demonstrates that the studied partial mitochondrial cytochrome oxidase I and II genes were found to be instrumental for the molecular identification of these forensically important flesh fly species.
昆虫学证据在法医案件的死亡时间推断中具有重要意义。麻蝇科(双翅目)在死亡时间估计中的应用受到限制,因为形态学鉴定常常因幼虫、蛹甚至成虫阶段的相似特征而受阻。为了使物种鉴定更加准确可靠,人们考虑采用基于DNA的鉴定方法。在本研究中,我们评估了利用线粒体细胞色素氧化酶I和II部分基因来鉴别来自埃及和中国的具有法医重要性的麻蝇科物种[银口麻蝇(Robineau-Desvoidy)、重毫麻蝇(Thomson)、白头麻蝇(Meigen)和努比亚尸食麻蝇(Wiedemann)]。使用聚合酶链反应扩增该区域,随后对扩增产物进行直接测序,并使用限制性内切酶HinfI和MfeI。使用Kimura双参数距离模型计算核苷酸序列差异,并生成邻接法系统发育树。所有检测的标本都被正确鉴定到物种。即使在属于麻蝇属的3个同域物种中,限制性内切酶HinfI和MfeI的组合也能提供不同的限制性片段长度多态性图谱。因此,本研究表明,所研究的线粒体细胞色素氧化酶I和II部分基因有助于对这些具有法医重要性的麻蝇物种进行分子鉴定。