Sara R. Bodnar MS, Coral Smith MS, Alekhlas A. Alsharji BS, Tina Moroose MS, Casper Venter PhD, Arati Iyengar PhD
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With GlobalFiler™, average autosomal STR (aSTR) profile completeness was 44.4% with 2–20 pg, 54.3% with 22–60 pg, and 95% with 64–250 pg DNA input. CODIS uploadable profiles were obtained in 2/10, 3/11, and 11/12 samples in these ranges. With ForenSeq™ DNA Signature Prep, average aSTR profile completeness was 19.7% with 1–20 pg and 45.2% with 22–47 pg but increased to 78.3% with 68–122 pg and 86.7% with 618–1000 pg DNA input. Uploadable profiles were obtained in 0/12, 4/11, 4/7, and 3/3 samples for these ranges. Results show very high sensitivity using both kits. Half-volume reactions and 30 cycles had minimal negative effect on Globalfiler™ profile quality, providing support for wider use after validation experiments to routinely improve results from LT samples. A standard protocol for the ForenSeq™ DNA Signature Prep kit was also highly successful with LT DNA obtained from CA-fumed fingerprints with additional information from isometric STR alleles and other markers.</p>","PeriodicalId":15743,"journal":{"name":"Journal of forensic sciences","volume":"69 5","pages":"1706-1717"},"PeriodicalIF":1.5000,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"DNA typing of cyanoacrylate fumed latent fingerprints using GlobalFiler™ and ForenSeq™ Signature Prep kits\",\"authors\":\"Sara R. Bodnar MS, Coral Smith MS, Alekhlas A. Alsharji BS, Tina Moroose MS, Casper Venter PhD, Arati Iyengar PhD\",\"doi\":\"10.1111/1556-4029.15566\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>DNA typing of latent fingerprints is highly desirable to increase chances of individualization. We recovered DNA from Cyanoacrylate (CA) fumed fingerprints and used both GlobalFiler™ and ForenSeq™ DNA Signature Prep kits for DNA typing. For GlobalFiler™, samples were processed using a protocol modified for Low Template (LT)-DNA samples (half-volume reactions, 30 cycles) while for ForenSeq™ DNA Signature Prep, samples were processed using a standard protocol and fluorometer-based library quantitation. We evaluated genotyping success and quality of profiles in terms of completeness, Peak Height Ratio/Allele Coverage Ratio, presence of PCR artifacts and drop-in alleles. With GlobalFiler™, average autosomal STR (aSTR) profile completeness was 44.4% with 2–20 pg, 54.3% with 22–60 pg, and 95% with 64–250 pg DNA input. CODIS uploadable profiles were obtained in 2/10, 3/11, and 11/12 samples in these ranges. 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引用次数: 0
摘要
对潜伏指纹进行 DNA 分型可增加个性化的机会。我们从氰基丙烯酸酯(CA)气相指纹中提取 DNA,并使用 GlobalFiler™ 和 ForenSeq™ DNA Signature Prep 试剂盒进行 DNA 分型。对于 GlobalFiler™,样本采用针对低模板(LT)-DNA 样本修改的方案进行处理(半量反应,30 个循环),而对于 ForenSeq™ DNA Signature Prep,样本采用标准方案和基于荧光计的文库定量进行处理。我们从完整性、峰高比/等位基因覆盖率、PCR伪影和等位基因丢失等方面评估了基因分型的成功率和图谱的质量。使用 GlobalFiler™,2-20 pg 的常染色体 STR (aSTR) 图谱平均完整率为 44.4%,22-60 pg 为 54.3%,64-250 pg DNA 输入为 95%。在这些范围内,分别有 2/10、3/11 和 11/12 份样本获得了 CODIS 可上传图谱。使用 ForenSeq™ DNA Signature Prep,1-20 pg 样品的平均 aSTR 图谱完整率为 19.7%,22-47 pg 样品的平均 aSTR 图谱完整率为 45.2%,而输入 68-122 pg DNA 样品的平均 aSTR 图谱完整率增至 78.3%,输入 618-1000 pg DNA 样品的平均 aSTR 图谱完整率增至 86.7%。在这些范围内,分别有 0/12、4/11、4/7 和 3/3 的样本获得了可上传的图谱。结果表明,这两种试剂盒的灵敏度都非常高。半量反应和 30 个循环对 Globalfiler™ 图谱质量的负面影响极小,支持在验证实验后更广泛地使用,以定期改进 LT 样品的结果。ForenSeq™ DNA Signature Prep 试剂盒的标准方案也非常成功地处理了从 CA 气相指纹中获得的 LT DNA,并从等距 STR 等位基因和其他标记物中获得了额外的信息。
DNA typing of cyanoacrylate fumed latent fingerprints using GlobalFiler™ and ForenSeq™ Signature Prep kits
DNA typing of latent fingerprints is highly desirable to increase chances of individualization. We recovered DNA from Cyanoacrylate (CA) fumed fingerprints and used both GlobalFiler™ and ForenSeq™ DNA Signature Prep kits for DNA typing. For GlobalFiler™, samples were processed using a protocol modified for Low Template (LT)-DNA samples (half-volume reactions, 30 cycles) while for ForenSeq™ DNA Signature Prep, samples were processed using a standard protocol and fluorometer-based library quantitation. We evaluated genotyping success and quality of profiles in terms of completeness, Peak Height Ratio/Allele Coverage Ratio, presence of PCR artifacts and drop-in alleles. With GlobalFiler™, average autosomal STR (aSTR) profile completeness was 44.4% with 2–20 pg, 54.3% with 22–60 pg, and 95% with 64–250 pg DNA input. CODIS uploadable profiles were obtained in 2/10, 3/11, and 11/12 samples in these ranges. With ForenSeq™ DNA Signature Prep, average aSTR profile completeness was 19.7% with 1–20 pg and 45.2% with 22–47 pg but increased to 78.3% with 68–122 pg and 86.7% with 618–1000 pg DNA input. Uploadable profiles were obtained in 0/12, 4/11, 4/7, and 3/3 samples for these ranges. Results show very high sensitivity using both kits. Half-volume reactions and 30 cycles had minimal negative effect on Globalfiler™ profile quality, providing support for wider use after validation experiments to routinely improve results from LT samples. A standard protocol for the ForenSeq™ DNA Signature Prep kit was also highly successful with LT DNA obtained from CA-fumed fingerprints with additional information from isometric STR alleles and other markers.
期刊介绍:
The Journal of Forensic Sciences (JFS) is the official publication of the American Academy of Forensic Sciences (AAFS). It is devoted to the publication of original investigations, observations, scholarly inquiries and reviews in various branches of the forensic sciences. These include anthropology, criminalistics, digital and multimedia sciences, engineering and applied sciences, pathology/biology, psychiatry and behavioral science, jurisprudence, odontology, questioned documents, and toxicology. Similar submissions dealing with forensic aspects of other sciences and the social sciences are also accepted, as are submissions dealing with scientifically sound emerging science disciplines. The content and/or views expressed in the JFS are not necessarily those of the AAFS, the JFS Editorial Board, the organizations with which authors are affiliated, or the publisher of JFS. All manuscript submissions are double-blind peer-reviewed.