摘要
目的建立15个常染色体和18个Y染色体STR基因座以及性别基因座的六色荧光标记复合PCR直扩检测体系,并评估其法医学应用价值。方法采用六色荧光标记技术,建立15个常染色体STR基因座(D3S1358、D13S317、D7S820、D16S539、TPOX、TH01、D2S1338、CSF1PO、D19S433、v WA、D21S11、D18S51、D8S1179、D5S818、FGA)和18个Y染色体STR基因座(DYS527a/b、DYS448、DYS456、DYS385a/b、DYS458、DYS391、DYS390、DYS19、DYS438、DYS393、DYS389Ⅰ、DYS439、DYS389Ⅱ、DYS392、GATA、DYS635)以及Amelogenin的复合扩增体系,收集800份无关人员血样进行基因座检测,评估所建复合扩增体系的稳定性、灵敏度、种属特异性、直扩可行性以及抗抑制性。结果本检测体系对800份无关人员血样复合扩增后,结果准确,灵敏度达0.125ng;种属特异性高;38份案件检材全部准确分型。在对照男性与女性的DNA浓度比值大于等于1:4时,男性DNA的所有基因座均能准确分型。若模板DNA中含一定浓度的已知抑制剂时,所有基因座也均准确分型。结论本文建立的复合扩增检测体系可同时检测15个常染色体和18个Y染色体基因座以及性别基因座,结果稳定准确,灵敏度高,可为法医DNA检测分析提供一个新选择。
Objective To develop a 6-dye fuorescence-labelling PCR multiplex protocol that directly amplifes 15 autosomal and 18 Y STR loci plus one Amelogenin gender indicator without DNA extraction, and evaluate its forensic applicability. Methods With 800 DNA samples of inter-irrelevant individuals, the developed protocol was tested by the samples to be amplifed locally at the 15 autosomal （D3S1358, D13S317, D7S820, D16S539, TPOX, TH01, D2S1338, CSF1PO, D19S433, vWA, D21S11, D18S51, D8S1179, D5S818, FGA） and 18 Y STR loci （DYS527a/b, DYS448, DYS456, DYS385a/b, DYS458, DYS391, DYS390, DYS19, DYS438, DYS393, DYS389Ⅰ, DYS439, DYS389Ⅱ, DYS392, GATA, DYS635） plus the gender marker Amelogenin. The protocol was evaluated on its stability, sensitivity, species specificity, viability and anti-inhibition. Results All the selected-34-locus STR profles were accurately and steadily amplifed from the 800 samples by the developed protocol, with the high species specifcity to be met and the detectable minimal DNA amount equal to or over 0.125ng. The 38 samples from real cases were correctly exposed of their genotypes. All the male STR loci had been exactly revealed when the control DNA was mixed with that of male and female in a ratio at or over 1： 4. Moreover, the anti-inhibition assay demonstrated that all the STR loci can be shown if the DNA template contains at or below certain concentrations of given PCR-inhibitors. Conclusion The developed protocol can be used to amplify the designed 15 autosomal and 18 Y STR loci plus the sex-typing marker Amelogenin with high portability, stability and sensitivity, capable of providing a new way for forensic DNA analysis.
出处
《刑事技术》
2018年第1期30-34,共5页
Forensic Science and Technology
基金
上海市科研计划项目(No.13JG0500200)
关键词
STR
复合PCR
直扩体系
六色荧光标记
法医遗传学
STR
multiplex PCR
direct amplifcation
6-dye fuorescence-labelling
forensic applicability
