扩增Amelogenin基因用于生物检材种属鉴定

目的扩增Amelogenin基因测定血痕或组织的种属,以确定在法医学检验中的应用价值。方法收集猪、羊、马等10几种常见动物的血痕或肌肉组织,应用PCR扩增Amelogenin基因,PAGE电泳,银染后观察结果。结果哺乳动物猪、牛、狗、羊、马、驴动物血痕扩增产物为1条带,片段长度102bp。猕猴检见106bp和112bp两条带,与人血痕没有区别。兔、猫、鼠血痕未检出特异性片段。其它常见物种鳝鱼、青蛙、鸭、鹅、鸽、鹌鹑、麻雀等均未见扩增产物。结论扩增Amelogenin基因进行种属鉴定,方法简单,灵敏度高,可应用于法医检案。     

Isolation of eugenol from biological material

The optimal conditions for isolation of eugenol from biological material by ethyl acetate were determined. Possible purification of the compound from coextractive substances of biomaterial on the colon with silica gel L 40/100 micro is demonstrated. Methods of thin-layer and high-yield liquid chromatography for identification and quantitative determination of eugenol are proposed.     

Isolation of lambda-cyhalothrin from biological material

Dioxan is proposed as an isolating agent for isolating lambda-cytalotrin (LC) from biological material. Optimal conditions for isolating LC from cadaveric hepatic tissue with dioxan were determined and the results of the isolation were quantified.     

Isolation of nitrobenzene from biological material

Toluene is recommended as an agent for extraction of nitrobenzene from biological material. Optimal conditions for nitrobenzene isolation from cadaveric human liver are defined and the results of isolation are quantitatively evaluated.     

Species identification from dried snake venom

Illegal trade in snake parts has increased enormously. In spite of strict protection under wildlife act, a large number of snakes are being killed ruthlessly in India for venom and skin. Here, an interesting case involving confiscation of crystallized dried snake venom and subsequent DNA-based species identification is reported. The analysis using the universal primers for cytochrome b region of the mitochondrial DNA revealed that the venom was extracted from an Indian cobra (Naja naja). On the basis of this report, the forwarding authority booked a case in the court of law against the accused for illegal hunting of an endangered venomous snake and smuggling of snake venom. This approach thus has immense potential for rapid identification of snake species facing endangerment because of illegal trade. This is also the first report of DNA isolation from dried snake venom for species identification.     

Characteristics of dipyridamol isolation from biological material

Acetone is proposed as an isolating agent for extraction of dipyridamolum from biological material. Optimal conditions of isolating dipyridamolum from human cadaveric liver tissue with acetone are determined and quantitative estimation of isolation results is provided.     

Characteristics of alphacypermethrin isolation from biological material

Methanol is proposed as an isolating agent for isolation of alphacypermethrin from biological material. Optimal conditions of isolating alphacipermetrin from human cadaveric liver tissue with methanol are determined and quantitative estimation of isolation results is provided.     

The identification investigation of the material evidence of biological origin after multifactorial exposure

The possibility of investigating pieces of material evidence of biological origin after exposure to various factors is evaluated. The possibility of detecting proteins of liquid media of human organism by electrophoresis in polyacrylamide denatured gel is investigated. The method is intended for identification of biological material in a state of grave destruction. Methodology of such studies is proposed. The data indicate that the structural integrity and qualitative composition of the spectrum of main serum proteins are retained after combined exposure to damaging factors and complete destruction of blood cells.     

扩增TP53内含子8用于生物检材的种属鉴定

目的扩增常见动物TP53内含子8片段,确定其在法医学生物检材种属鉴定中的应用价值。方法收集标本为包括人在内的15种常见动物的血痕或肌肉组织,提取DNA后定量,应用PCR扩增TP53内含子8,PAGE电泳,银染后观察结果。结果人和猕猴都扩增出一条长度为460bp的片段,鳝鱼、鲢鱼、青蛙、鸭、兔、猫、小白鼠、豚鼠、猪、牛、羊虽有扩增产物,但不在分型区内,鸡、狗未见扩增产物。结论扩增TP53内含子8进行种属鉴定,方法简单,灵敏度较高。     

Purification of extracts from biological material using aluminum oxide

An effective, rapid, and safe operation is suggested for purification of extracts from biological material from co-extractive substances during testing for barbiturates, benzophenones, and nichlofolane. The purification is carried out on a column packed with aluminum oxide.     

混合血迹中不同个体成份逐一分离识别的研究

目的研究法庭科学混合血迹物证中不同个体成份逐一分离、识别的问题,建立适合混合血迹个体识别的分析技术。方法采用PCR-SSCP及测序技术,选择m tDNA D-loop区的HVI 16030~16481区域452 bp片段作为分析目标,对中国汉族两无关个体、三无关个体混合血迹进行分析。结果100份两个体混合血迹样品m tDNA 452bp的PCR产物经SSCP电泳分离,结果有95份样品完全分离开,分离成功率达95%;30份三个体混合血迹样品452 bp片段经SSCP电泳分离,结果有26份样品有1~3个个体完全分离开,分离成功率达84%。对其中3份两个体混合血样、2份三个体混合血样SSCP电泳分离后的谱带进行回收、测序分析,两个体混合血样每一份均可准确获得其中单一个体序列及以另一个体主要成份(峰值比达4∶1以上)的序列结果;三个体混合血迹中不同个体成份可以达到初步分离,1份可准确确定单一个体序列。对两个体不同比例混合样品SSCP分析,结果可以检测到较少成份的最低比例为20∶80。结论本研究建立的PCR-SSCP及测序分析混合血迹综合技术,是对混合血迹中不同个体成份逐一分离、识别的一种有效技术手段。     

Sex identification of white-tailed deer using frozen venison

Fluids from frozen muscle tissue taken from 449 white-tailed deer (Odocoileus virginianus) were analyzed using a radioimmunoassay technique for testosterone. Comparing 309 deer sampled in early November, antlered males had significantly higher tissue testosterone levels (range = 0.18 to 16.00 ng/mL) than females (range = 0.00 to 0.18 ng/mL) while antlerless (fawn) males overlapped both groups (range = 0.00 to 0.68 ng/mL). Samples taken from 132 deer from January through August showed no differences among ages or sexes. Antlered males can be differentiated from females during the breeding season.     

DNA profiles from biological trace material contained in underwear and toothbrushes

There are special cases where it is extremely difficult to find a close relative for obtaining reference samples, such as in adoption cases or foreigners without relatives in the country. In these cases, it is possible to use personal objects from the missing person. We developed a protocol for obtaining DNA profiles from toothbrushes and underwear and evaluated two DNA extraction methods (Silica and magnetic particles), comparing the DNA quantity and the quality of the obtained profiles. Results showed that for underwear, silica in suspension recovered higher DNA quantity but magnetic particles (PrepFiler®) generated more complete profiles. For toothbrushes PrepFiler® recovered a higher DNA quantity and generated more complete DNA profiles. Statistical analyses showed there were no significant differences between both methods. Higher DNA quantities were recovered from underwear samples, however toothbrushes generated higher quality in DNA profiles, without obtaining mixed profiles, concluding that toothbrushes are a better item than underwear. Considering that silica in suspension method is three-times cheaper than PrepFiler®, we propose it is possible to successfully obtain DNA profiles from personal objects using a low-cost DNA extract method.