Sperling F A, Anderson G S, Hickey D A
Department of Biology, University of Ottawa, Ontario, Canada.
J Forensic Sci. 1994 Mar;39(2):418-27.
Insect larvae found on a corpse can be used for estimating postmortem intervals. Here, we describe a molecular method for rapid identification of these insects. Specific insect DNA fragments were amplified using the polymerase chain reaction (PCR), followed by direct DNA sequencing of the amplification products. We sequenced 2300 base pairs of mitochondrial DNA from each of three blowfly species: Phormia regina, Phaenicia sericata and Lucilia illustris. All three species are important in forensic entomology. We found 118 nucleotide differences between the L. illustris and P. sericata sequences, 186 between L. illustris and P. regina, and 192 between P. sericata and P. regina. Based on these abundant DNA sequence differences, we can unambiguously identify the immature larval stages of these insects. These DNA sequence differences were also used to predict species-specific, diagnostic restriction sites in the amplified DNA, and these predictions were verified by digestion with nine restriction enzymes. The DNA sequences reported here encode the mitochondrial COI, COII and tRNA-leucine genes.
在尸体上发现的昆虫幼虫可用于估计死后间隔时间。在此，我们描述一种用于快速鉴定这些昆虫的分子方法。使用聚合酶链反应（PCR）扩增特定昆虫DNA片段，随后对扩增产物进行直接DNA测序。我们对三种丽蝇的线粒体DNA各2300个碱基对进行了测序，这三种丽蝇分别是：黑须亚麻蝇（Phormia regina）、丝光绿蝇（Phaenicia sericata）和亮绿蝇（Lucilia illustris）。这三个物种在法医昆虫学中都很重要。我们发现亮绿蝇和丝光绿蝇序列之间有118个核苷酸差异，亮绿蝇和黑须亚麻蝇之间有186个差异，丝光绿蝇和黑须亚麻蝇之间有192个差异。基于这些丰富的DNA序列差异，我们能够明确鉴定这些昆虫的未成熟幼虫阶段。这些DNA序列差异还被用于预测扩增DNA中物种特异性的诊断性限制性酶切位点，并且通过用九种限制性酶进行酶切验证了这些预测。此处报道的DNA序列编码线粒体细胞色素氧化酶亚基I（COI）、细胞色素氧化酶亚基II（COII）和亮氨酸转运RNA基因。
