密实骨和骨小梁中的 DNA 保存

IF 3.2 2区 医学 Q2 GENETICS & HEREDITY Forensic Science International-Genetics Pub Date : 2024-06-01 DOI:10.1016/j.fsigen.2024.103067
Irena Zupanič Pajnič, Nika Kovačič
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引用次数: 0

摘要

紧密骨和骨小梁的分子结构存在显著差异。在密实骨中,骨粉需要完全溶解才能有效地从羟基磷灰石中释放 DNA。而在保留了软组织的骨小梁中,我们认为不需要完全溶解骨粉就能从胶原蛋白中释放 DNA。为了研究这个问题,我们对 45 对第二次世界大战期间的干骺端(密实骨)-干骺端(骨小梁)股骨进行了研究,每对股骨都采用了完全溶解(FD)和部分溶解(PD)提取方法。使用 qPCR PowerQuant 分析评估了 DNA 的质量和数量,并对常染色体 STR 进行了分型,以确认分离 DNA 的真实性。我们的研究结果支持密实骨和骨小梁中不同的 DNA 保存机制,因为仅在密实骨中,FD 方法比 PD 方法更有效,而在骨小梁中未观察到 DNA 产量的差异,这表明在处理骨小梁组织时无需完全溶解骨粉。此外,在应用 PD 法时,紧密骨和骨小梁的 DNA 产量有显著差异,骨小梁提取的 DNA 多于紧密骨。在对密实骨和骨小梁进行处理时,用 FD 方法分离出的 DNA 数量相似,这也证明了用 PD 方法处理骨小梁非常合适。此外,用 FD 法提取密实骨和用 PD 法提取骨小梁时,分离出的 DNA 数量也相似。在对骸骨进行常规鉴定时,用 PD 法处理骨小梁可缩短提取过程并简化研磨过程。
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DNA preservation in compact and trabecular bone

Significant variation exists in the molecular structure of compact and trabecular bone. In compact bone full dissolution of the bone powder is required to efficiently release the DNA from hydroxyapatite. In trabecular bone where soft tissues are preserved, we assume that full dissolution of the bone powder is not required to release the DNA from collagen. To investigate this issue, research was performed on 45 Second World War diaphysis (compact bone)–epiphysis (trabecular bone) femur pairs, each processed with a full dissolution (FD) and partial dissolution (PD) extraction method. DNA quality and quantity were assessed using qPCR PowerQuant analyses, and autosomal STRs were typed to confirm the authenticity of isolated DNA. Our results support different mechanisms of DNA preservation in compact and trabecular bone because FD method was more efficient than PD method only in compact bone, and no difference in DNA yield was observed in trabecular bone, showing no need for full dissolution of the bone powder when trabecular bone tissue is processed. In addition, a significant difference in DNA yield was observed between compact and trabecular bone when PD was applied, with more DNA extracted from trabecular bone than compact bone. High suitability of trabecular bone processed with PD method is also supported by the similar quantities of DNA isolated by FD method when applied to both compact and trabecular bone. Additionally similar quantities of DNA were isolated when compact bone was extracted with FD method and trabecular bone was extracted with PD method. Processing trabecular bone with PD method in routine identification of skeletonized human remains shortens the extraction procedure and simplifies the grinding process.

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来源期刊
CiteScore
7.50
自引率
32.30%
发文量
132
审稿时长
11.3 weeks
期刊介绍: Forensic Science International: Genetics is the premier journal in the field of Forensic Genetics. This branch of Forensic Science can be defined as the application of genetics to human and non-human material (in the sense of a science with the purpose of studying inherited characteristics for the analysis of inter- and intra-specific variations in populations) for the resolution of legal conflicts. The scope of the journal includes: Forensic applications of human polymorphism. Testing of paternity and other family relationships, immigration cases, typing of biological stains and tissues from criminal casework, identification of human remains by DNA testing methodologies. Description of human polymorphisms of forensic interest, with special interest in DNA polymorphisms. Autosomal DNA polymorphisms, mini- and microsatellites (or short tandem repeats, STRs), single nucleotide polymorphisms (SNPs), X and Y chromosome polymorphisms, mtDNA polymorphisms, and any other type of DNA variation with potential forensic applications. Non-human DNA polymorphisms for crime scene investigation. Population genetics of human polymorphisms of forensic interest. Population data, especially from DNA polymorphisms of interest for the solution of forensic problems. DNA typing methodologies and strategies. Biostatistical methods in forensic genetics. Evaluation of DNA evidence in forensic problems (such as paternity or immigration cases, criminal casework, identification), classical and new statistical approaches. Standards in forensic genetics. Recommendations of regulatory bodies concerning methods, markers, interpretation or strategies or proposals for procedural or technical standards. Quality control. Quality control and quality assurance strategies, proficiency testing for DNA typing methodologies. Criminal DNA databases. Technical, legal and statistical issues. General ethical and legal issues related to forensic genetics.
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