血痕氨水涂膜热解离方法的改进

探讨热解离法在不同条件下的实验效果。对已知血型的血痕用改进的氨水涂膜热解离法进行 ABH血型物质测定 ,结果显示检材在 4℃冰箱中吸收 15min左右、 56℃恒温箱中解离 10min后再加入指示红细胞继续解离 5min观察效果最好。此法简单易行，适用于基层单位。     

微量血痕AB型粘附法检验

微量血痕用胶纸粘附-热解离法检验ABH血型,其方法基本同于吸附-热解离法,经改进后有操作简便、节省检材、缩短检验时间、可携带至现场直接提取、又可多次利用检验的优点。     

应用血膜热解离法检测体液斑ABO血型

血膜热解离法检验体液斑中的ABO型物质，具有操作简单、检验时间短、灵敏、特异性高等优点，而且与中和法相比具有节省检材的特点，在实际检验中取得了较好的效果。材料和方法一、实验材料1.已知血型的人唾液斑、精斑、阴道分泌物斑纱布各1（）份（从医院提取）；2抗A、抗B血清（淮北市公安局血清室提供）；3‘）型血痕纱布（采耳血涂在白绷带纱布上，室温保存，最长为1年）。二、实验方法（一）血膜热解离法检测体液斑ABt）血型l取ZCC长唾液斑、精斑、阴道分泌物斑纱线加入本理盐水3滴浸泡半小时或充分搅拌，取二满分别满于载被片两侧…     

不同吸收温度与时间对热解离法检验血型结果的影响

本文在血痕浓度等项恒定的基础上,用不同的吸收温度和时间,分别对已知A、B、O、AB型血痕进行了热解离法检验,并对检测结果进行了观察、分析、比较,从中找出了最佳的吸收温度和时间,为法医物证热解离法检验提供了捷径。     

人体组织ABO血型检测方法的研究

探讨人体组织ABO血型检测方法。对已知ABO血型尸体的不同组织,用红细胞粘连试验、吸收—抑制试验和吸收－解离试验进行ABO血型测定。12例尸体的16种组织中均检出与尸体血痕相同的ABH物质。对不同温度保存的组织块进行ABH物质检测的结果显示,4℃保存的组织ABH物质的检出时间长于室温,空腔脏器的检出时间短于实质脏器。三种方法中,红细胞粘连试验简单易行,适用于基层单位,吸收－抑制试验用于组织块的测定时优于吸收－解高试验。     

室外毛巾上腐败血痕STR分析1例报道

1材料与方法1.1案例2006年某日,深圳市某山顶发现一具高度腐败女尸,现场提取一块方形小毛巾(有灰黑色可疑血痕),送检做DNA检测。1.2方法1.2.1提取DNA方法一:常规Chelex-100法[1]提取方形小毛巾上可疑血痕。方法二:应用德国QIAGEN公司生产的BioRo-bot EZ1全自动核酸纯化仪[2](以下简称BioRobotEZ1)提取方形小毛巾上可疑血痕。(1)剪取适量检材,置0.5 ml Eppendorf管中,加入EZ1试剂盒(EZ1 DNA Tissue Kit(48)(Cat.No.953034))中试剂BufferG2 190μl,PK 10μl,56℃裂解30 min,去载体将裂解液转移置2.0 ml样本管中。(2)在BioRobo…     

红细胞酸性磷酸酶(EAP)型的分布及血痕EAP的检出

本文应用琼脂糖凝胶电泳法对辽宁地区213例汉族随机献血员的红细胞酸性磷酸酶(EAP)型进行了检测,其基因频率为EAP~(?)=0.169,EAP~b-0.831。结合文献资料分析了EAP型分布的种族差异,指出了各人种EAP型分布的特点。用琼脂糖凝胶电泳法,对红细胞溶血液EAP型的最小检出量为2μL(2×3mm滤纸条)及0.5μL(直接加样);对血痕EAP型的最小检出量为7.5μL血液制成的检样。本法在4℃保存4周以内的红细胞溶血液和室温保存25天的血痕均能正确判定EAP的型别。     

血痕Gm(2)因子检验及分布

应用抗 Gm(2)血清和抗 D-Gm(2)血清以及 Gm 标准对照血清,选择新鲜 ORh(D)红细胞,用凝集抑制试验,对北京地区269人血清或新鲜血痕进行检验。结果 Gm(2+)分布为29%,Gm(2-)分布为71%。对已确定 Gm(2)型的230份血痕,在室温放置14个月后再次检验,结果与第一次所测血型完全相同。同时对11起案件中的血痕检验了 Gm(2)因子,起到了排除嫌疑人或增加与案件相关联的作用。     

单克隆抗M、抗N抗体的研制及初步应用

用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10～15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性和高效价抗 M 和抗 N 抗体的细胞株,并建立了 GM_4H3、GM_4H_4、N_2A_3和 N_2D_(10)细胞株。这些细胞株可持续分泌免疫球蛋白 G 类抗 M、抗 N 抗体。应用这些抗体,通过血凝法、解离法和 ELISA 斑点法,可进行血及血痕的 MN 分型。在血型检验中,优于多克隆抗 M、抗N 血清。     

样品成份对“咖啡环”法DNA检测的影响

目的调查样品中常见成份对"咖啡环"法DNA检测的影响。方法将含有不同浓度SDS、Na Cl等的DNA-EB滴加到载玻片上,室温自然蒸发,用凝胶成像系统观察荧光沉积形貌(Deposition Pattern,DP)、计算积分光密度(Integrated Optical Density,IOD),分析量效关系。结果低浓度非DNA成份存在时,DP仍以环状沉积为主,但有年轮环、放射纹等成份特异性细微差异。浓度高时,DP因成份而异,可为环+散在光点型(Na Cl)、中央光斑+弱小环型(SDS)、同心或年轮状多环型(Triton X-100)、环+斑点型(H^+)等。非DNA成份对DNA含量估计有不同程度影响,但估计值多数在0.5~2倍以内。结论样品成份对DNA的DP和IOD均有影响,但不影响DNA含量的粗略估计,且能提供样品纯度和残留物信息。     

保存温度与时间对生物样品中氯胺酮稳定性的影响

目的研究不同温度和时间条件保存时生物样品中氯胺酮的稳定性。方法家兔以氯胺酮150mg/kg灌胃,30min后处死,取其血、肝、肾、脑,分别在室温（18～24℃）和冷冻（-20℃）条件下保存,并用气相色谱-质谱法定性分析、气相色谱-氮磷检测器法测定不同时间各样品中氯胺酮含量。结果血、肝、肾、脑冷冻保存至第30天氯胺酮含量均降低（P〈0.05）;室温条件下各样品中氯胺酮含量自第5天起均升高（P〈0.05）。结论生物样品在冷冻条件下保存时氯胺酮稳定性较好,怀疑氯胺酮中毒或死亡的检材应冷冻保存,尽快检测。     

Alteration of the Death Scene After Self‐stabbing: A Case of Sharp Force Suicide Disguised by the Victim as a Homicide?

This paper reports a case of a 72‐year‐old woman who was found dead in her bedroom with a 4 cm vertical stab wound in the abdomen. A bloodstained knife was found in the top drawer of her bedside table. The clothes worn by the victim showed no damage. A bloodstained vest and a sweater with frontal incisions were found far from the victim, in the bathroom and in the bedroom respectively. Several bloodstains were found in every room of the apartment. The evidence found during the forensic examination and, in particular, the Bloodstain Pattern Analysis, led the investigators to determine the manner of death, being consistent with a suicide with a long‐lasting physical activity after self‐stabbing. This report describes an unusual case of “disguised suicide,” in which the victim tried to cover‐up the suicide by changing her clothes and concealing the weapon, in the last minutes of her life.     

The analysis of biological samples from crime scene for a future human DNA profile confrontation. Effects of presumptive test reagents on the ability to obtain STR profiles for human identification

Because of the increase of evidence of blood stains, that have been washed or cleaned in an attempt to mask the analysis of DNA profiles, there is also an increase in the use of presumptive tests on samples sent to laboratories. Some of the presumptive tests, used to identify blood and semen stains, could potentially affect the recovery of high molecular weight DNA from the samples, or extinguish them, especially those already present in small quantities. After the presumptive tests, often these samples are discarded. This study aimed to examine the possibility of obtaining a DNA profile from samples submitted for presumptive testing and cleaned with bleaches with and without chlorine. Two different protocols were conducted: (a) A unique sample of human blood in natura (5 μL), already typed through the DNA techniques with the genetic profile previously known (control), was distributed onto cotton fabrics and dried at room temperature. Four samples of fabric were macerated in saline solution and Coombs serum and then stored for three months (room temperature and freezer −20 °C). (b) Another sample of human blood, type A, in natura, already typed through the techniques of DNA (control) was used. Aliquots of 200 μL were distributed in: cotton, denim and synthetic fabric. The samples were dried at room temperature for 24 h. The blood stains in those fabrics (cotton, denim and synthetic) were then divided into three groups: unwashed, cleaned with chlorine bleach and cleaned with chlorine bleach and soap powder. The samples were again dried at room temperature for 24 h, before the use of luminol. The DNA were extracted with Chelex 100 and amplified with the Identifiler Kit (Applied Biosystems). The blood stains exposed to saline and Coombs serum had DNA profiles consistent with untreated samples (controls). This result shows that the experts should keep and store the samples treated with saline and Coombs serum for future DNA confrontation when necessary. Also discussed in this paper the pattern of blood stains after washing with bleaching solutions, as well as the quantity of DNA obtained from these samples.     

The effect of luminol on presumptive tests and DNA analysis using the polymerase chain reaction.

This study was designed to test the following factors involved with processing luminol treated bloodstained evidence: 1) The reactivity of other presumptive chemical color tests, phenolphthalin (PT) and tetramethylbenzidine (TMB), following the application of the light emitting luminol presumptive test. 2) The effect of different cleanings of various bloody substrates on the luminol test. 3) The effect of different cleanings of various bloody substrates on the ability to obtain DNA suitable for PCR testing. 4) The ability to extract DNA from luminol treated bloodstained substrates using three extraction techniques. 5) The effect of spraying washed and unwashed bloodstains on various substrates with luminol on the ability to correctly type the DNA using PCR. Our findings indicated that luminol did not adversely effect the PCR testing and did not interfere with the PT and TMB presumptive tests for blood. It was determined that the substrate and the method of cleaning were the major factors affecting DNA yield and the ability to type the bloodstains using PCR based technologies.     

Micromethod for MN antigen grouping of dried bloodstains

A micromethod based on the absorption elution technique was shown to be applicable to the detection of M and N blood groups of dried bloodstains on cotton cloth. Each antigen M and N was tested using two different types of antisera. Two hundred different bloodstains, stored up to six months, were analyzed. Conclusive results were obtained for M typing on 2.5-mm-long bloodstained threads. For N typing, some cross-reactivity of homozygous M stains with anti-N sera was observed. This may be explained by the structure of the M and N antigens on the red cell membrane.     

基于超低温环境下的DFO荧光手印增强技术

目的研究超低温环境下DFO荧光手印增强技术。方法采用二级复迭式机械制冷，在超低温环境下增加DFO显现手印的荧光强度。结果温度对荧光物质的荧光强度有显著影响，当温度降至-40℃以后荧光强度随激发波长的加长而加强。结论本技术方法对常用纸张上DFO显现的手印荧光效果明显。     

广州人群红细胞GLOI表型的分布调查及血痕GLOI的检测

本文报道了广州地区人群的GLOI表型分布,基因频率为:GLOI~1=0.1716,GLOI~2=0.8284。室温下(25-30℃)保存的血痕20天内可全部正确分型;日晒8小时,露天过夜,4℃保存105天的血痕均可正确分型;流水冲洗2小时的血痕不能分型;室内腐败血标本9天内检测结果可靠。在实际应用时应考虑这些因素。     

氯胺酮在急性中毒大鼠体内的死后再分布研究

目的探索氯胺酮在大鼠体内的死后再分布变化规律及温度对再分布的影响。方法48只雄性SD大鼠随机分为2个实验组（室温组24只、冷藏组18只）和1个对照组（6只）,实验组大鼠以氯胺酮290mg／kg灌胃,45min后缺氧处死,分别置于室温（24℃）和冷藏（4℃）条件下,于死后不同时间（0、12、24、48h）取心血、外周血、肝、肺、肾、心肌、大脑,检测其中氯胺酮含量;对照组大鼠以生理盐水灌胃,各对应组织器官样品为空白对照。血和组织样品中加入内标物SKF。。后碱化,乙酸乙酯萃取,GC／MS全扫描定性,内标法、工作曲线法气相色谱定量分析。结果室温条件下,大鼠死后48h内随着死亡时间延长,心血、肺、肝中氯胺酮的浓度呈升高趋势（P〈0．05）,肾脏中氯胺酮的浓度先升高后下降（P〈0．05）,外周血、心肌和脑中氯胺酮的浓度无显著性变化（P〉0．05）。冷藏条件下,血液及组织中氯胺酮浓度变化无显著性差异（P〉0．05）,除心肌外,各样本浓度均低于相应时段室温条件保存的样本。结论氯胺酮在大鼠体内存在死后再分布现象。温度对大鼠死后血液及组织中氯胺酮浓度变化有较明显的影响。     

The simultaneous electrophoretic analysis of esterase D and phosphoglucomutase subtyping in fresh blood and in dried bloodstains

Phosphoglucomutase1 (PGM) subtyping and esterase D phenotyping were simultaneously performed by electrophoresis of bloodstained fibers using agarose and a Tris-maleic acid buffer system , pH 5.4. This method reduces anodal gel shrinkage and shortens development time when compared to the conventional electrophoretic technique for PGM subtyping which is performed at pH 7.4 using an agarose-starch substrate.     

龙胆紫染色法在显微捕获单细胞分离技术中的应用

目的建立用于显微捕获单细胞技术的龙胆紫染色法并评价其应用效果。方法制作20枚口腔拭子脱落细包悬液,配制0．05g／mL龙胆紫染液。取100μL口腔细胞悬液加入0．15、0．25、0．5、1．0、2．0μL染色液,考察最佳染色浓度;争别染色5、10、30、60min,考察最佳染色时间;用最佳条件染色后抓捕3个细胞,采用联合LV-PCR技术扩增并进行DNA分驯佥测,进行重复试验20次,并对同一检材未经染色的抓捕细胞进行检测,用于比对STR分型成功率。将龙胆紫染色法用于案例检材的口腔上皮细胞分离检验。结果1001μL细胞悬液加入0．5止0．05g／mL龙胆紫染液,染色5min,胞核呈紫工色,与胞浆对比明显;染色时间延长不影响染色效果及细胞分离检验结果。优化后的龙胆紫染色法对低体积扩增无归显抑制（P〉0．05）。应用此染色法于案例检材,胞核标识清晰,STR分型结果达同一认定标准。结论龙胆紫染色法刚于细胞核的发现,有助于提高显微捕获单细胞技术的检测效能。