利用MicroGEM的forensicGEM DNA提取技术从降解的911样本中检测出DNA
2023-05-16 来源:本站 点击次数:1190
2022年10月19日--来自佩斯大学、纽约市立约翰杰刑事司法学院和纽约市首席医疗官办公室（OCME）的研究小组今天公布了使用forensicGEM Universal从世贸中心袭击事件的高度降解样本中提取DNA的发现。这些发现在法医科学家东北部协会（NEAFS）2022年年会上的法医生物学/DNA部分做了介绍。
自2001年9月11日的袭击以来，OCME已经努力确认了近20,000具遗体。而从尚待鉴定的遗体中提取核酸被公认为是最具挑战性的工作，因为大部分的DNA已经由于暴露在火、热、飞机燃料、自然元素和老化而降解。
使用forensicGEM试剂盒，研究人员能够从世贸中心的样本中提取DNA，而此前使用传统的提取方法是无法检测到DNA的。
典型的法医DNA提取行业标准使用蛋白酶K，需要多次离子清洗，孵化时间长，并且需要大量的原始样本。ForensicGEM方法具有显著的优势。使用新型的“Cocktail(鸡尾酒)”嗜热酶提取方法，提取过程中不使用洗涤剂和还原剂，forensicGEM提供的DNA提取过程不需要纯化步骤，因此可以从非常少量的样本中快速提取DNA。此外，减少的处理可以保护样品的完整性。
研究人员首先比较了forensicGEM的孵化时间、酶量、骨骼制备方法和提取后的纯化，并将其与使用非世贸中心骨骼样本的传统提取方法进行了比较。
ForensicGEM协议在80%的骨质样本中检测到了DNA，而这些骨质样本以前没有检测到DNA。然后，forensicGEM被应用于世贸中心袭击事件的五个实际样本，以前没有从这些样本中提取到可检测的DNA，从这五个样本中的三个提取到了DNA，并在这些高度降解的样本中的另外两个样本产生了一个22-locus和一个15-locus的图谱。鉴于ForensicGEM的样品要求（10毫克）比传统的提取（2克）要少得多，骨骼提取的成功是值得关注的。
研究小组表示，他们未来的研究将调查刮骨与标准的研磨方法，因为这项研究使用刮骨方法取得了更高的DNA数量和更好的轮廓。
"MicroGEM的法医专家和应用科学家Jillian Conte博士说："这项研究为推动围绕骨质样本DNA提取的科学发展迈出了新的一步，允许进行以前不可能完成的鉴定。"我们很高兴与处理这些困难样本类型的研究小组合作，利用我们独特的嗜热提取能力。
MicroGEM在其最近推出的Sal6830 COVID-19诊断仪中使用了类似的核酸提取方法，这是世界上第一个在医疗点检测SARS-CoV-2的基于唾液的PCR测试，它在30分钟内得到结果。
MicroGEM的法医产品，包括forensicGEM Universal和forensicGEM Sperm，是从血液、组织、骨骼、唾液和精子等珍贵、有限的法医样本中提取DNA的理想选择。
Extraction of Challenging Forensic Samples Using the MicroGEM DNA Extraction Kit
Lauren Chwatt, Pace University, Falyn Vega, John Jay College of Criminal Justice, James Prinston & Andrew Schweighardt, NYC Office of Chief Medical Examiner
DNA extraction is an essential but sometimes tedious process in forensic investigation that may require a significant investment of time and resources. Proteinase K has been an industry standard for DNA extraction for several decades due to its reliability of protein denaturation when performing an extraction. Some of the drawbacks of proteinase K are that its use requires multiple ionic detergents and washing steps, while only being active above 65 ̊C. Here, we analyze the potential of a new enzyme being used in DNA extraction known as forensicGEM by the manufacturer MicroGEM. This novel enzyme is temperature-dependent, which enables it to be compatible with mesophilic enzymes. The forensicGEM protocol offers complete DNA extraction in about 20 minutes in a single tube, thus limiting contamination, loss of sample, and working time -- ultimately increasing efficiency. One of the main attractions of forensicGEM is its ability to extract DNA from highly degraded samples, potentially leading to more complete STR profiles in samples where a profile may have previously been poor or unattainable by conventional extraction procedures. To assess the efficiency and potential uses of forensicGEM, we collected highly degraded tissue and bone samples and extracted DNA using the MicroGEM kit, altering different parameters such as incubation times, enzyme amount, bone preparation method, and post-extraction purification. We then compared the results of samples extracted with MicroGEM to the results of the same samples extracted with a standard organic extraction to assess whether this new technology could be utilized routinely on highly degraded samples. Half of the degraded samples extracted with MicroGEM had detectable DNA. The highest success rate was observed for bone samples. One tissue sample in particular yielded higher average peak heights when extracted with MicroGEM. No statistically significant pattern was apparent with respect to identifying superior MicroGEM optimization parameters. Success with bone profiling was notable given that there was much less sample input for MicroGEM (10 mg) compared to the organic extraction (2 g). An ancillary finding of this study is that the bone reparation method of scraping yielded higher DNA quantities and better quality profiles compared to samples treated with the standard method of milling. Since the initial results were promising, this new technology was utilized on remains from the 9/11 World Trade Center attacks from which no detectable DNA had been previously extracted. Ultimately, MicroGEM was able to yield a 22-locus and a 15-locus profile on two of these highly degraded samples. Future work will focus on further investigation of the bone scraping method for universal application, and continued optimization of experimental parameters in the MicroGEM extraction protocol.
自2001年9月11日的袭击以来，OCME已经努力确认了近20,000具遗体。而从尚待鉴定的遗体中提取核酸被公认为是最具挑战性的工作，因为大部分的DNA已经由于暴露在火、热、飞机燃料、自然元素和老化而降解。
使用forensicGEM试剂盒，研究人员能够从世贸中心的样本中提取DNA，而此前使用传统的提取方法是无法检测到DNA的。
典型的法医DNA提取行业标准使用蛋白酶K，需要多次离子清洗，孵化时间长，并且需要大量的原始样本。ForensicGEM方法具有显著的优势。使用新型的“Cocktail(鸡尾酒)”嗜热酶提取方法，提取过程中不使用洗涤剂和还原剂，forensicGEM提供的DNA提取过程不需要纯化步骤，因此可以从非常少量的样本中快速提取DNA。此外，减少的处理可以保护样品的完整性。
研究人员首先比较了forensicGEM的孵化时间、酶量、骨骼制备方法和提取后的纯化，并将其与使用非世贸中心骨骼样本的传统提取方法进行了比较。
ForensicGEM协议在80%的骨质样本中检测到了DNA，而这些骨质样本以前没有检测到DNA。然后，forensicGEM被应用于世贸中心袭击事件的五个实际样本，以前没有从这些样本中提取到可检测的DNA，从这五个样本中的三个提取到了DNA，并在这些高度降解的样本中的另外两个样本产生了一个22-locus和一个15-locus的图谱。鉴于ForensicGEM的样品要求（10毫克）比传统的提取（2克）要少得多，骨骼提取的成功是值得关注的。
研究小组表示，他们未来的研究将调查刮骨与标准的研磨方法，因为这项研究使用刮骨方法取得了更高的DNA数量和更好的轮廓。
"MicroGEM的法医专家和应用科学家Jillian Conte博士说："这项研究为推动围绕骨质样本DNA提取的科学发展迈出了新的一步，允许进行以前不可能完成的鉴定。"我们很高兴与处理这些困难样本类型的研究小组合作，利用我们独特的嗜热提取能力。
MicroGEM在其最近推出的Sal6830 COVID-19诊断仪中使用了类似的核酸提取方法，这是世界上第一个在医疗点检测SARS-CoV-2的基于唾液的PCR测试，它在30分钟内得到结果。
MicroGEM的法医产品，包括forensicGEM Universal和forensicGEM Sperm，是从血液、组织、骨骼、唾液和精子等珍贵、有限的法医样本中提取DNA的理想选择。
Extraction of Challenging Forensic Samples Using the MicroGEM DNA Extraction Kit
Lauren Chwatt, Pace University, Falyn Vega, John Jay College of Criminal Justice, James Prinston & Andrew Schweighardt, NYC Office of Chief Medical Examiner
DNA extraction is an essential but sometimes tedious process in forensic investigation that may require a significant investment of time and resources. Proteinase K has been an industry standard for DNA extraction for several decades due to its reliability of protein denaturation when performing an extraction. Some of the drawbacks of proteinase K are that its use requires multiple ionic detergents and washing steps, while only being active above 65 ̊C. Here, we analyze the potential of a new enzyme being used in DNA extraction known as forensicGEM by the manufacturer MicroGEM. This novel enzyme is temperature-dependent, which enables it to be compatible with mesophilic enzymes. The forensicGEM protocol offers complete DNA extraction in about 20 minutes in a single tube, thus limiting contamination, loss of sample, and working time -- ultimately increasing efficiency. One of the main attractions of forensicGEM is its ability to extract DNA from highly degraded samples, potentially leading to more complete STR profiles in samples where a profile may have previously been poor or unattainable by conventional extraction procedures. To assess the efficiency and potential uses of forensicGEM, we collected highly degraded tissue and bone samples and extracted DNA using the MicroGEM kit, altering different parameters such as incubation times, enzyme amount, bone preparation method, and post-extraction purification. We then compared the results of samples extracted with MicroGEM to the results of the same samples extracted with a standard organic extraction to assess whether this new technology could be utilized routinely on highly degraded samples. Half of the degraded samples extracted with MicroGEM had detectable DNA. The highest success rate was observed for bone samples. One tissue sample in particular yielded higher average peak heights when extracted with MicroGEM. No statistically significant pattern was apparent with respect to identifying superior MicroGEM optimization parameters. Success with bone profiling was notable given that there was much less sample input for MicroGEM (10 mg) compared to the organic extraction (2 g). An ancillary finding of this study is that the bone reparation method of scraping yielded higher DNA quantities and better quality profiles compared to samples treated with the standard method of milling. Since the initial results were promising, this new technology was utilized on remains from the 9/11 World Trade Center attacks from which no detectable DNA had been previously extracted. Ultimately, MicroGEM was able to yield a 22-locus and a 15-locus profile on two of these highly degraded samples. Future work will focus on further investigation of the bone scraping method for universal application, and continued optimization of experimental parameters in the MicroGEM extraction protocol.
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