Chen L, Sun G, Wu M
Hua Xi Yi Ke Da Xue Xue Bao. 1994 Sep;25(3):253-8.
Sex identification of human tooth is an important aspect of individual identification in forensic science practice. This study was designed to determine the effects of environmental and physicochemical factors (EPF) on the deoxyribonucleic acid (DNA) from dental pulp and PCR result. Extracted teeth were subjected to the following treatment varying pH (2, 7, 10) solution; 10% formalin, 75% ethanol; different temperature (-20 degrees C, 4 degrees C, 25 degrees C, 37 degrees C, boiling, burning); humidity (20%, 66%, water submergence); burying the teeth outdoors; and aging (teeth stored at room temperature for 1.5, 3.5 and 8 years). DNA was extracted from the tooth followed by agarose gel electrophoresis for the purpose of DNA assay and quantity determination. Amplification was carried out using a pairs of primer Y1.1 and Y1.2. The results of the study showed that, 154 bp fragment was obtained in all male teeth, whereas no 154 bp fragment was found in a female fresh tooth. 154 bp fragment bands were not observed in case of teeth subjected to burial outdoors for 10 weeks, submergence in water for 10 weeks, soak in pH 2 solution for 10 weeks, and combustion for 10 minutes; only faint bands were observed in case of teeth subjected to 66% humidity and pH 10 solution for 10 weeks and combustion for 5 minutes; obvious bands were observed in the rest cases. In addition, the quality of DNA from dental pulp affected PCR amplification result. The integrity of DNA isolated was categorized as being high-molecular-weight (HMW), HMW with partial degraded DNA, completely degraded DNA, completely degraded and not detected.(ABSTRACT TRUNCATED AT 250 WORDS)
人类牙齿的性别鉴定是法医学实践中个体识别的一个重要方面。本研究旨在确定环境和物理化学因素(EPF)对牙髓脱氧核糖核酸(DNA)及聚合酶链反应(PCR)结果的影响。提取的牙齿接受以下处理:不同pH值(2、7、10)的溶液;10%福尔马林、75%乙醇;不同温度(-20℃、4℃、25℃、37℃、煮沸、燃烧);湿度(20%、66%、水浸);户外掩埋牙齿;以及老化(牙齿在室温下保存1.5年、3.5年和8年)。从牙齿中提取DNA,随后进行琼脂糖凝胶电泳以进行DNA检测和定量测定。使用一对引物Y1.1和Y1.2进行扩增。研究结果表明,所有男性牙齿均获得154 bp片段,而女性新鲜牙齿中未发现154 bp片段。在户外掩埋10周、水浸10周、浸泡在pH值为2的溶液中10周以及燃烧10分钟的牙齿中未观察到154 bp片段条带;在湿度为66%和pH值为10的溶液中处理10周以及燃烧5分钟的牙齿中仅观察到 faint条带;在其余情况下观察到明显条带。此外,牙髓DNA的质量影响PCR扩增结果。分离出的DNA完整性分为高分子量(HMW)、部分降解的HMW DNA、完全降解的DNA、完全降解且未检测到。(摘要截短于250字)