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对储存在乙二胺四乙酸(EDTA)、氟化钠/草酸钾和无添加剂试管中的死后全血进行DNA回收的纵向评估。

Longitudinal assessment of DNA recovery from post-mortem whole blood stored in EDTA, sodium fluoride/potassium oxalate and additive-free tubes.

作者信息

Grobbelaar Jana, Vuko Loyiso Abongile Marvin, Davies Bronwen, Pearce Brendon, Musiyandaka Fungisai Lorraine, Heathfield Laura Jane

机构信息

Division of Forensic Medicine and Toxicology, Department of Pathology, Faculty of Health Sciences, University of Cape Town, Anzio Road, Cape Town, South Africa.

Genetics Department, Faculty of Agriscience, Stellenbosch University, Van Der Bijl Street, Stellenbosch, South Africa.

出版信息

Int J Legal Med. 2025 Mar;139(2):495-507. doi: 10.1007/s00414-024-03384-z. Epub 2024 Dec 10.

Abstract

Adverse drug reactions and fatalities can result from therapeutic drug use due to genetic deficiencies in drug-metabolizing enzymes. In cases where ancillary testing may not reveal a clear cause of death, molecular autopsies can be valuable. However, forensic mortuaries do not retain DNA samples in all cases, which hinders subsequent genetic testing if it is later deemed necessary. This study aimed to evaluate whether post-mortem whole blood samples collected for toxicological analysis, could provide viable DNA for genetic testing after varying storage periods. Thirty deceased individuals were recruited with informed consent. Blood collected at autopsy from each individual was stored in two sodium fluoride/potassium oxalate (gray-top) tubes, two additive-free (red-top) tubes and one ethylenediaminetetraacetic acid (EDTA; purple-top) tube- the latter recommended for DNA analysis. Blood from one gray-top and one red-top tube were sampled for toxicological analysis prior to DNA analysis, while the remaining samples (acting as controls) underwent DNA analysis immediately. DNA analysis involved DNA extraction and DNA concentration and degradation assessment. Blood samples were stored at 4 °C and DNA extraction and analysis was repeated one year and then five years later. Toxicological sampling did not significantly influence DNA results. DNA concentration and quality significantly decreased over time for all sample types, with DNA from red-top tubes showing the greatest decline. The study showed that DNA testing for drug-metabolizing enzymes was feasible on whole blood that had been stored for five years. This finding supports the potential for retrospective genetic testing in cases of adverse drug reactions and fatalities.

摘要

由于药物代谢酶的遗传缺陷,治疗性药物使用可能导致药物不良反应和死亡。在辅助检测可能无法揭示明确死因的情况下,分子尸检可能会有帮助。然而,法医停尸房并非在所有情况下都保留DNA样本,如果之后认为有必要进行基因检测,这会阻碍后续检测。本研究旨在评估为毒理学分析采集的死后全血样本在不同储存时间后,是否能提供用于基因检测的可用DNA。在获得知情同意的情况下招募了30名死者。从每个个体尸检时采集的血液分别储存在两支含氟化钠/草酸钾(灰色帽)试管、两支无添加剂(红色帽)试管和一支乙二胺四乙酸(EDTA;紫色帽)试管中——后者推荐用于DNA分析。在进行DNA分析之前,从一支灰色帽试管和一支红色帽试管中采集血液进行毒理学分析,而其余样本(作为对照)立即进行DNA分析。DNA分析包括DNA提取以及DNA浓度和降解评估。血液样本储存在4°C,一年后再五年后重复进行DNA提取和分析。毒理学采样对DNA结果没有显著影响。所有样本类型的DNA浓度和质量均随时间显著下降,红色帽试管中的DNA下降最为明显。该研究表明,对储存五年的全血进行药物代谢酶的DNA检测是可行的。这一发现支持了在药物不良反应和死亡病例中进行回顾性基因检测的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/815f/11850465/a823341d587a/414_2024_3384_Fig1_HTML.jpg

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