共查询到17条相似文献,搜索用时 343 毫秒
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用引物Y_3、Y_4和PCR方法鉴定性别的法医学应用 总被引:1,自引:0,他引:1
用Y3、Y4和Alu9.1、Alug.2两对引物和PCR方法检测陈旧血痕和毛根的性别获得成功。引物Y3、Y4扩增的靶序列位于Y染色体特异3.4Kb重复序列中,扩增产物为460bp;引物Alu9.1、Alu9.2用以扩增男女共有的Alu重复序列,扩增产物为130bP。室温保存13年之久的19例脐带血血痕(男性9冽,女性10例)和室温保存10~11个月的10例已知性别自然脱落毛根(男性6例,女性4例)的性别测定结果均正确;对一起凶杀案的血痕性别测定为定案提供了重要证据。本方法简化了样品的前处理过程。 相似文献
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作者用引物Y_3、Y_4和DNA聚合酶链式反应(PCR)作微量人类血液(痕)和毛根的性别鉴定。扩增的靶序列位于Y染色体DNA特异3.4kb重复序列中,扩增产物为460bp。检材用量为:新鲜血液0.5μl、血痕纱纤维1mm、毛根单个。20例保存4个月的血痕与2例保存6年半的血痕性别判定结果均正确,无性别记载的保存9~11年的3例血痕显现了清晰的460bpY特异DNA扩增带。15例保存20天的自然脱落毛根性别判定结果均正确。本法省略了检材处理中的酚-氯仿抽提DNA等纯化步骤,既简化了实验操作,又减少了检验过程中外源DNA的污染机会和样品DNA的损耗,使这一性别鉴定方法更符合法医学实践的需要。 相似文献
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应用 PCR 技术同时扩增人 ZFY 和 ZFX 基因特异的 DNA 序列,在男性血痕中可检测到两种扩增产物,即340bp 长的 ZFY 基因及488bp 长的 ZFX 基因特异 DNA 片段;在女性血痕中仅可检测到488bp 长的 ZFX 基因特异 DNA 片段,据此判定干血痕性别。干血痕的最小检出需要量为0.125μl 血液量的血痕。室温保存10年的血痕可以准确判定性别。ZFY 基因位于 Y 染色体短臂。本方法同时检测两条性染色体,可以避免由于扩增失败或 Y 染色体长臂变异出现的假阴性或假阳性。扩增产物经琼脂糖凝胶电泳即可区分。 相似文献
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目的 建立一种采用PCR技术对降解DNA样本进行性别鉴定的新方法。方法 采用针对amelogenin基因X染色体外显子3bp缺失设计的引物AMELU1及AMELD1,对在室温环境下放置5-15年的男、女血痕标本各50例、毛发各20例、骨骼各20例以及现场提取5--20天的男、女腐败肌肉各10例标本中提取的降解DNA样本进行扩增。用PGA(9%T,3%C)电泳、银染显带检测扩增产物。结果 所有样本均得到正确结果,男性检材表现为83bp的Y特异性及80bp的X特异性2条谱带,而女性检材仅有1条80bp的X特异性谱带。结论 用针对amelogenin基因X染色体外显子3bp缺失设计的引物AMELU1及AMELD1鉴定性别的方法灵敏、可靠、方便,是降解DNA检材性别鉴定十分理想的方法。 相似文献
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目的 建立一种采用PCR技术对降解DNA样本进行性别鉴定的新方法。 方法 采用针对amelogenin基因X染色体外显子 3bp缺失设计的引物AMELU1及AMELD1,对在室温环境下放置 5~ 15年的男、女血痕标本各 5 0例、毛发各 2 0例、骨骼各 2 0例以及现场提取 5 - - 2 0天的男、女腐败肌肉各 10例标本中提取的降解DNA样本进行扩增。用PAG( 9%T ,3 %C)电泳、银染显带检测扩增产物。 结果 所有样本均得到正确结果 ,男性检材表现为 83bp的Y特异性及 80bp的X特异性 2条谱带 ,而女性检材仅有 1条 80bp的X特异性谱带。 结论 用针对amelogenin基因X染色体外显子 3bp缺失设计的引物AMELU1及AMELD1鉴定性别的方法灵敏、可靠、方便 ,是降解DNA检材性别鉴定十分理想的方法。 相似文献
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扩增X—Y同源Amelogenin基因内含子在性别鉴定中的应用研究 总被引:2,自引:1,他引:1
本文用一对针对X-Y同源的Amelogenin基因第一内含子的引物,于同一试管中分别扩增出针对于x和Y染色体的特异性DNA片段:106bp及112bp的PCR产物,经聚丙烯酰胺凝胶电泳后用银染法观察扩增结果,取得了满意的效果.对50pgDNA模板室温放置16年的血痕及单根毛发、烟头等检材均可得到明确的性别鉴定结果.本实验证明该方法简便、灵敏、可靠、特别适用于腐败降解检材的法医学性别鉴定. 相似文献
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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。 相似文献
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ABO genotyping by polymerase chain reaction. 总被引:10,自引:0,他引:10
ABO blood group system's genotyping by polymerase chain reaction in genomic DNA level is developed. The positions of nucleotide 258 and 700 of cDNA from A transferase were used to distinguish A, B, and O alleles by restriction enzyme digestion. To identify the 258th nucleotide, a 199- or 200-bp DNA fragment was amplified by PCR and digested with Kpn I. For the 700th nucleotide, a 128-bp PCR amplified fragment was designed and digested with Alu I. By examining the DNA fragment digested patterns, ABO genotypes were easily determined. Results obtained using this method on 20 ABO-known peripheral blood samples showed that this new technique could provide accurate ABO genotype. Biologic forensic samples, such as, blood stains, saliva stains, semen stains, hair, bone tissue, and semen contaminated with vaginal secretion were also successfully typed. This rapid, sensitive and reliable method should be applicable not only in forensic identification but also in medical examination. 相似文献
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A recombinant DNA probe hybridizing specifically to human repeat DNA sequence (pHY10) of which about 3000 copies are present on the Y chromosome was used for sex determination of degraded DNA samples of blood stains. Human blood stains of male and female origin were readily differentiated with the pHY10 DNA probe. This radioactive technique enabled reliable and sensitive sex determination from blood or dried blood stains greater than 20 years old. Less than 1 microliter of blood or 1 piece of 0.5 cm length thread of blood stain from cotton fabric was sufficient for the test using dot blot hybridization. Compared with the radioactive labeling method, the photobiotin labeling method showed one thirtieth to one fiftieth lower sensitivity and presented some problems which are expected to be resolvable. 相似文献
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应用‘Myo’小卫星 DNA 探针,Southern 印迹杂交技术,对血斑、精斑、同一个体不同组织进行 DNA 指纹图分析,均获得清晰的图谱。同一个体的血斑与血液、精斑与精液以及不同的组织其 DNA 指纹图谱完全相同。可以根据斑痕或组织与嫌疑个体的血液或某一组织 DNA 的指纹图谱比对以做出同一认定。50μl 血液量的血斑、5μl 精液量的精斑可以获得清晰易辨的指纹图谱。五年的精斑、两年的血斑亦可做出与同源个体新鲜精液、血液完全一致的 DNA 指纹图谱。对杀人、强奸杀人、碎尸等不同案件的血痕、精斑、不同组织碎块进行了 DNA 指纹图检验,均做出了正确的个体认定。本方法的应用为我国法医物证检验提供了新的分析手段,使个体认定得以实现。 相似文献
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Male DNA typing from 25-year-old vaginal swabs using Y chromosomal STR polymorphisms in a retrial request case. 总被引:3,自引:0,他引:3
We report here the application of Y chromosomal DNA analysis in a retrial request case, raised officially by Sapporo High Court, Japan, of a condemned criminal whose capital punishment has been suspended. DNA was extracted from mixed seminal/vaginal secretion stains collected 25 years ago from two raped and murdered victims, and Y chromosome STR loci (DYS19, 390, 393, YCAII) were amplified and sequenced to clarify the DNA type of the rapist. Alkaline proteinase and sodium hydroxide were used before phenol/chloroform extraction to achieve high quality DNA from very old samples. In addition, amplified fragments of DYS19, DYS390, and DYS393 were sequenced using an automated DNA sequencer. Four Y STR DNA types detected from vaginal swabs were found identical to those of the accused criminal and confirmed that the two rape and murder cases had been committed by the same person. Sapporo High Court accepted the results and rejected the retrial request in February 1998. 相似文献
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本文报道了对98例中国人 pMCT118位点扩增片段长度多态性的等位基因频率调查及有关法医生物物证检验问题的研究。用多聚酶链反应、小型聚丙烯酰胺凝胶电泳和银染方法,在12h 内检测低至1ngDNA 的 pMCT118位点扩增片段长度多态性。已发现22个等位基因,大小分布在340~780bp 之间,基因频率为0.005~0.3,杂合度为79%。对6个家系22名相关个体的分析符合孟德尔定律。探讨了微量检材样品制备和对混合斑、单根毛囊、唾液等生物检材的法医应用。对20个实际案例进行了检验。 相似文献
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Recombinant DNA hybridizing specifically to a 300 nucleotide repeat DNA sequence (BLUR8) of human specificity and to human repeat DNA sequence (pHY10) on the Y chromosome was used for human identification and sex determination of degraded DNA samples of blood stains, dental pulp, and bone marrow. This radioactive technique enabled reliable and sensitive human and sex determination from blood stains that were more than 80 years old. Less than 1 piece of 0.5 cm length thread of blood stain was enough for both tests. DNA from relatively fresh dental pulp and bone marrow was clearly identified. The human identification test, which could recognize up to 0.3 ng DNA correctly, was 3 to 5 times more sensitive than the sex determination test. 相似文献