Cattaneo C, DiMartino S, Scali S, Craig O E, Grandi M, Sokol R J
Istituto di Medicina Legale e delle Assicurazioni, Università degli Studi di Milano, Italy.
Forensic Sci Int. 1999 Jun 28;102(2-3):181-91. doi: 10.1016/s0379-0738(99)00059-6.
In situations where badly burnt fragments of bone are found, identification of their human or non-human origin may be impossible by gross morphology alone and other techniques have to be employed. In order to determine whether histological methods were redundant and should be superseded by biomolecular analyses, small fragments of artificially burnt bone (human and non-human) were examined by quantitative and standard light microscopy, and the findings compared with newer biomolecular analyses based on identifying specific human albumin by ELISA and amplifying human mitochondrial DNA by PCR. For quantitative microscopy, reference data were first created using burnt bones from 15 human and 20 common domestic and farm animals. Measured osteon and Haversian canal parameters were analysed using multivariate statistical methods. Highly significant differences were found between values for human and non-human bone, and a canonical discriminant function equation was derived, giving a predicted correct classification of 79%. For the main study, samples of cortical bone were taken from three fresh cadavers, six human skeletons and ten freshly slaughtered animals and burnt by exposure to temperatures ranging from 800 to 1200 degrees C; charred fragments of human cortical bone from two forensic cases were also tested. Quantitative microscopy and canonical discriminant function gave the correct origin of every sample. Standard microscopy falsely assigned burnt bone from one human skeleton and one forensic case to a non-human source, but otherwise gave correct results. Human albumin was identified in five individuals, including one of the forensic cases, but mitochondrial DNA could not be amplified from any of the human bone. No false positive test results were seen with either biomolecular method; and human albumin and mitochondrial DNA were correctly identified in all unburnt control specimens. It was concluded that histological methods were not redundant and that quantitative microscopy provided an accurate and consistent means of determining the human or non-human origin of burnt bone and was more reliable than standard microscopy or the newer immunological and DNA techniques tested here.
在发现严重烧焦的骨碎片的情况下，仅通过大体形态学可能无法确定其人类或非人类来源，因此必须采用其他技术。为了确定组织学方法是否多余，是否应被生物分子分析所取代，对人工烧焦的骨碎片（人类和非人类）的小碎片进行了定量和标准光学显微镜检查，并将结果与基于酶联免疫吸附测定法鉴定特定人类白蛋白和通过聚合酶链反应扩增人类线粒体DNA的更新的生物分子分析结果进行比较。对于定量显微镜检查，首先使用来自15个人类以及20种常见家畜和农场动物的烧焦骨头创建参考数据。使用多元统计方法分析测量的骨单位和哈弗斯管参数。发现人类和非人类骨头的值之间存在高度显著差异，并得出了一个典型判别函数方程，预测正确分类率为79%。在主要研究中，从三具新鲜尸体、六具人类骨骼和十只刚宰杀的动物身上采集皮质骨样本，并通过暴露在800至1200摄氏度的温度下进行焚烧；还测试了来自两起法医案件的人类皮质骨烧焦碎片。定量显微镜检查和典型判别函数给出了每个样本的正确来源。标准显微镜检查错误地将一具人类骨骼和一起法医案件中的烧焦骨头归为非人类来源，但其他情况下结果正确。在五个人中鉴定出了人类白蛋白，包括一起法医案件中的一个，但无法从任何人类骨头中扩增出线粒体DNA。两种生物分子方法均未出现假阳性检测结果；并且在所有未焚烧的对照标本中均正确鉴定出了人类白蛋白和线粒体DNA。得出的结论是，组织学方法并非多余，定量显微镜检查提供了一种准确且一致的方法来确定烧焦骨头的人类或非人类来源，并且比标准显微镜检查或此处测试的更新的免疫和DNA技术更可靠。
