汗潜指印的STR分型检测

目的探索汗潜指印的荧光STR复合扩增检测的方法。方法采用Chelex-100和Microco-100浓缩柱,提取汗潜指印中DNA,STR复合扩增荧光电泳检测。结果 105例汗潜指纹STR分型可明确判读5个以上基因座的占30.3％,个体之间的差异、捺印指印时用力大小以及指印遗留在客体上时间的长短均影响检测成功率。结论该汗潜指印的DNA提取方法步骤简单,方法较为稳定,使单枚汗潜指印可望获得DNA分型。     

汗潜指印的STR分型检测在案件中的应用

目的探索案例中所涉及的汗潜指印DNA的提取和检验方法。方法采用Chlex-100法提取DNA,进行STR复合扩增,通过毛细管电泳检测荧光信号。结果案例中涉及的汗潜指印在ProfilerPlus试剂盒10个基因座的分型检测均获成功。结论含有汗潜指印的检材的发现和正确提取对最终检测成功至关重要。     

3种提取胶带粘面汗潜指印中DNA的方法比较

目的比较胶带粘面汗潜指印中DNA提取的方法。方法分别采用硅珠法、QIAMicrokit法、硅珠-QIAMicrokit法提取胶带粘面的汗潜指印中DNA,STR复合扩增,荧光电泳检测。结果用QIAMicrokit法、硅珠-QIAMicrokit法提取胶带粘面汗潜指印中DNA,检测成功率分别为21%和36%。硅珠法检测未获成功。结论硅珠-QIAMicrokit法提取胶带粘面汗潜指印中的DNA比QIAMicrokit法,检验时间更短,检测成功率更高。     

8种方法显现的汗潜指印STR分型研究

目的研究常见指印显现方法对指印STR检验的影响。方法采用Invisorb spin forensic试剂盒提取纯化人汗潜指印DNA,低拷贝模板(LCN)STR复合扩增,荧光电泳检验。结果用铜粉、铝粉、荧光粉、黑磁粉、"502"胶、茚三酮、磺酸双三嗪荧光显色液显现的玻片、纸张和胶带纸粘面上的汗潜指印可成功进行STR分型。结论常见指印显现方法不影响指印STR检验。     

“502”熏显后掌纹的STR检验2例

随着STR基因座复合扩增系统在法庭科学中的广泛应用，汗潜指印的STR分型检验已成为可能。本文对2枚“502”熏显后的掌纹进行了STR检验，现报道如下。     

短波紫外照射对汗潜手印DNA检测的影响初探

目的探测短波紫外灯的照射是否会对汗潜手印DNA检测产生影响。方法由每名志愿者在纸张上捺印4枚拇指指印使每枚指印的脱落细胞量保持基本相同,抽取每名志愿者所捺印的一枚指印作为一组,共有四组,将三组指印置于短波紫外灯的照射下,照射时间分别设置为10min﹑30min和1h,还有一组不照射,然后用磁珠法对所有指印提取DNA并进行定量。结果短波紫外灯的照射会对汗潜手印中的DNA造成减损,照射时间越长,减损得越多。结论尽量减少短波紫外灯对汗潜手印照射的时间(可控制在10min内),以保证汗潜手印有足够量的DNA而能被用于STR分型检测。     

茚三酮显现汗潜指印的三种操作方法对DNA检测的影响

目的探讨并比较茚三酮显现汗潜指印的3种操作方法,即溶液浸泡法、涂抹法和喷雾法对后续DNA检测带来的影响。方法取16名志愿者的指印分4组,分为茚三酮浸泡法、涂抹法和喷雾法以及空白组,然后提取DlNA进行定量和STR分型检测。结果3种操作方法都会减少汗潜指印DNA的量,喷雾法的损失量最大,浸泡法和涂抹法结果比较接近。结论现场可疑指印检材,应根据检验需要决定指印显现和DNA提取的先后顺序。     

不同染色组织切片及不同组织固定方法对检测STR基因座的影响

目的探讨不同染色组织切片及不同组织固定方法对DNA检验的影响。方法采用Chelex100法及浓缩纯化方法提取DNA,PCR扩增后310型遗传分析仪检测。结果福尔马林固定4天以内或70%乙醇、无水乙醇分别固定1年及15年的HE染色切片可以检见Amel及9个STR基因座。PTAH等5种特殊染色未能检出相应基因座DNA谱带。结论70%乙醇或无水乙醇固定组织,石腊包埋组织及HE染色切片可以作为DNA?STR检验鉴定的样本材料。     

小偏角定向反射法拍摄汗液指印1例

2002年11月25日凌晨,武汉市汉阳区一出租车司机被杀,现场勘查发现出租车内后排右侧座位扶手上有一枚汗潜指印。该扶手为浅灰色塑料,表面较粗糙,指印遗留处微有弧度。指印为汗潜指印,侧光观察隐约可见,但与背景客体之间反差极弱。拍摄方法:在暗室内采用小偏角定向反射配光检验(偏角控制在5°左右),通过取景器观察并调整光源位置,至最佳效果后拍照固定,提取到一枚反差良好,具备鉴定条件的指印(图1、2)。配光原理:扶手表面粗糙,呈漫反射性质。指印纹线物质相对光滑,呈混合反射性质,而混合反射光在与入射角相等的反射角方向上强度最大。因此,指…     

DNA提取方法和PCR循环次数对STR扩增成功率的影响

目的探讨常见载体上的微量血痕DNA的提取方法、PCR循环次数对STR扩增成功率的影响。方法分别应用Chelex-100法及Chelex-100结合纯化法对8种载体上不同大小的血痕样本进行DNA提取,并采用28次、30次及34次PCR循环进行STR扩增,分别观察其扩增成功率。结果经28次、30次及34次PCR循环,Chelex-100法提取DNA后的STR扩增成功率分别为0.2917,0.3333,0.4583,Chelex-100结合纯化法的STR扩增成功率分别为0.3750,0.4583,0.8750。结论用Chelex-100结合纯化法提取DNA,用34次循环扩增可提高STR基因座的检测成功率。     

Evaluation of DNA Extraction Methods for Processing Fingerprint Powder-Coated Forensic Evidence

In unison, fingerprinting and DNA analysis have played a pivotal role in forensic investigations. Fingerprint powders that are available on the market can come in a range of colors and with specific properties. This study evaluated the efficiency of DNA extraction from samples coated with 3 brands of fingerprint powders: Lightning, Sirchie, and SupraNano, covering a range of colors and properties. A total of 23 fingerprint powders were tested using the Chelex, Promega DNA IQ™, and Applied Biosystems™ PrepFiler™ DNA extraction protocols. The DNA IQ™ and PrepFiler™ methods extracted higher yields of DNA in comparison to Chelex, which also accounted for better quality of PowerPlex x00AE; 21 DNA profiles recovered. There were no signs of degradation or inhibition in the quantification data, indicating that samples returning low DNA yield was due to interference during DNA extraction and not PCR inhibition. DNA profiles were recovered from the majority of fingerprint powders with only a single powder, Sirchie Magnetic Silver, failing to produce a profile using any of the methods tested. A link was observed between the DNA extraction chemistry, fingerprint powder property, that is, nonmagnetic, magnetic and aqueous, and the brand of fingerprint powder. Overall, the DNA IQ™ method was favorable for nonmagnetic fingerprint powders, while magnetic fingerprint powders produced more DNA profiles when extracted with the PrepFiler™ chemistry. This study highlights the importance of screening DNA extraction chemistries for the type of fingerprint powder used, as there is not a single DNA extraction method that suits all fingerprint powder brands and properties.     

Chelex法和两种磁珠法提取接触DNA效果的比较

目的比较Chelex法、DNA IQ磁珠法、EQ国产磁珠法对接触DNA的提取效果。方法将稀释为10ng、100ng的标准品DNA,分别采用Chelex法、DNA IQ磁珠法、EQ国产磁珠法处理;对30例烟蒂和30例牙刷分别采用Chelex法、DNA IQ磁珠法和EQ国产磁珠法提取DNA,然后进行PCR定量和STR检测。结果Chelex法对DNA的提取无损失,DNA IQ磁珠法、EQ国产磁珠法对DNA的提取均有不同程度的损失;烟蒂、牙刷等检材采用Chelex法提取的接触DNA量和IPC CT值显著高于IQ磁珠法、EQ国产磁珠法,但STR检验成功率却低于IQ磁珠法、EQ国产磁珠法。2种磁珠法提取的DNA量、IPC CT值和STR检验成功率无显著性差异。结论污染轻、杂质少的接触DNA检材,用Chelex法提取最为方便快捷;IQ磁珠法、EQ国产磁珠法更适合污染接触DNA检材的提取及自动化操作。     

Chelating resin-based extraction of DNA from dental pulp and sex determination from incinerated teeth with Y-chromosomal alphoid repeat and short tandem repeats

A procedure utilizing Chelex 100, chelating resin, was adapted to extract DNA from dental pulp. The procedure was simple and rapid, involved no organic solvents, and did not require multiple tube transfers. The extraction of DNA from dental pulp using this method was as efficient, or more so, than using proteinase K and phenol-chloroform extraction. In this study, the Chelex method was used with amplification and typing at Y-chromosomal loci to determine the effects of temperature on the sex determination of the teeth. The extracted teeth were incinerated in a dental furnace for 2 minutes at 100 degrees C, 200 degrees C, 300 degrees C, 400 degrees C, and 500 degrees C. After the isolation of DNA from the dental pulp by the Chelex method, alphoid repeats, and short tandem repeats, the human Y chromosome (DYZ3), DYS19, SYS389, DYS390, and DYS393 could be amplified and typed in all samples incinerated at up to 300 degrees C for 2 minutes. The DYS389 locus in some samples could not be amplified at 300 degrees C for 2 minutes. An autopsy case is described in which genotypings of DYS19, DYS390, and DYS393 from dental pulp obtained from a burned body were needed. The data presented in this report suggest that Chelex 100-based DNA extraction, amplification, and typing are possible in burned teeth in forensic autopsy cases.     

用Chelex100法及有机溶剂提取法联合提取DNA扩增STR基因座的比较研究

为了比较不同方法提取DNA扩增STR基因座的成功率 ,本文用Chelex10 0法及Chelex10 0法联合有机溶剂提取法分别提取性犯罪案件 6份血斑及 6份混合斑中精子DNA ,并用PE公司ProfilerPlus系统复合扩增 ,ABI 310型基因分型仪检测。结果表明 ,常规采用Chelex10 0法提取血斑及精斑等生物检材DNA作STR基因座检验失败或所检测的位点较少时 ,应对Chelex10 0法提取的DNA上清液再用有机溶剂提取法提取 ,可极大提高DNA检验成功率。采用本文建立的Chelex10 0法联合有机溶剂提取法 ,提高了PCR扩增阳性率 ,对实际检案很有帮助 ;在PCR扩增前应常规进行DNA定量检验 ,否则对于重要检材应进行梯度扩增。     

微量口腔脱落细胞检材的DNA检验

目的寻求提高微量口腔脱落细胞检材的DNA检验成功率的简便有效的提取方法。方法对不同载体上的100份微量口腔脱落细胞检材采用小体积Chelex-100法提取DNA,在ABI7500型荧光定量PCR仪上进行定量,同时用IdentifilerTM复合扩增系统扩增,在ABI3130遗传分析仪上进行STR分型。结果从25根饮料吸管上提取的DNA量在0.72～116.7.8ng,16个水杯杯缘提取的DNA量在2.15-142.5ng,31个饮料瓶(罐)口提取的DNA量在1～34.65ng,10根筷子上提取的DNA量在3.35～26.6ng,12个果核中提取的DNA量在0.294～21.4ng,6份吃剩的骨头中提取的DNA量在0.88～5.88ng。100份检材性别及9个以上STR位点分型成功率平均为59.38%。除了使用者的个人原因外,检材的提取送检方式、检材的质地、饮料的性质对提取的DNA量有显著影响,是否加蛋白酶K对提取的DNA量无显著影响。结论采用小体积Chelex-100法可对60%左右的微量口腔脱落细胞检材提取DNA进行STR分型。     

成都汉族群体10个STR基因座的遗传多态性

目的获得成都汉族群体10个短串联重复序列（short tandem repeat,STR）基因座的群体遗传学数据,评估其法医学应用价值。并对其种属特异性进行研究。方法Chelex法提取100名无亲缘关系的成都汉族个体血液样本的DNA,PCR扩增,非变性聚丙烯酰胺凝胶电泳,硝酸银显色分型。选择10种常见的动物样本进行种属特异性研究,对上述10个基因座的种属特异性进行评价。结果D1S2145、D3S2433、D5S1507、D5S2502、D8S2319、D9S926、D16S767、D17S2181、GATA140E03和GATA196B10基因座在成都汉族群体中等位基因个数分别为6、5、8、5、6、7、7、5、7和7,基因型个数分别为17、14、28、15、16、18、15、14、19和21;等位基因和基因型频率分布均符合Hardy-Weinberg平衡定律。经与10种动物样本比较,D3S2433、D5S1507、D5S2502、D8S2319和CATA196B10有较好的种属特异性,其余基因座分型区内少部分动物有扩增产物。结论上述10个STR基因座在成都汉族群体中具有较好的遗传多态性,在个体识别和亲权鉴定中具有一定的应用价值。     

Y-chromosomal short tandem repeats haplotyping from vaginal swabs using a chelating resin-based DNA extraction method and a dual-round polymerase chain reaction

Reported are 2 autopsy cases in which Y-chromosomal microsatellite short tandem repeats DYS19, DYS389I and II, DYS390, and DYS393 could be haplotyped with vaginal swabs by using a Chelex 100-based DNA extraction method and dual-round polymerase chain reaction. The extraction of DNA from vaginal swabs by using this method was as efficient or more efficient than using proteinase K and phenol-chloroform extraction or the alkaline lysis methods. Y-chromosomal microsatellite short tandem repeats haplotyping based on the dual-round polymerase chain reaction method provided genotypes from all the loci determined. Although amplification of Y-chromosomal microsatellite short tandem repeats loci is not directly involved in the existence of spermatozoa, it is considerably advantageous for male individualization from body fluid mixture stains in criminal cases.     

p33.4位点扩增片段长度多态性及其法医学应用的研究

以聚合酶链反应（PCR）、聚丙烯酰胺凝胶垂直电泳和银染法对中国100名无关个体小卫星区域p33．4位点的扩增片段长度多态性（Amp－FLPs）进行了研究，检出了8个等位基因。通过BIOTRAC系统进行数据处理．各等位基因重复单位的数目分别为7、10到15，其中在13～14之间发现一差值不足一个重复单位长度的罕见等位基因。片段长度分布于603～1115bP之间，基因频率分布于0.5～33．5％间，杂合度为64％，DP值为84．5％。对5个家庭25名相关个体进行分析，符合孟德尔遗传定律；对同一个体不同组织的DNA进行P33.4位点的分型研究表明，该技术适宜于法医物证检验。此外，本研究以Chelex处理不同检材制备DNA模板用于扩增，率先建立了比常规方法更快、更简便、更为实用的检验方法。     

烟蒂DNA分型的研究

目的 研究烟蒂中DNA提取及其检验。方法 用Chelex-100法提取170枚烟蒂样本的DNA,进行PCR扩增及STR检验。结果 除1名志愿者提供的21枚烟蒂外层纸未能检出STR基因分型外,其余烟蒂外层纸均得到分型结果。加入少许烟丝的样本未能检出STR分型,与口唇接触的海绵有时可检出基因型,6个月内的烟蒂可检出小片段基因座。结论 烟蒂能进行DNA分型,在法医检案中具有应用价值。