Evaluation of five commercial nucleic acid extraction kits for their ability to inactivate Bacillus anthracis spores and comparison of DNA yields from spores and spiked environmental samples
Evaluation of five commercial nucleic acid extraction kits for their ability to inactivate Bacillus anthracis spores and comparison of DNA yields from spores and spiked environmental samples
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DOI：
10.1016/j.mimet.2008.09.004
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发表时间：
2009-01-01
影响因子：
2.2
通讯作者：
Bowen, Michael D.
中科院分区：
文献类型：
--
作者：
Dauphin, Leslie A.;Moser, Benjamin D.;Bowen, Michael D.
本研究评估了五种商业提取试剂盒从炭疽芽孢杆菌孢子和加标环境样品中回收 DNA 的能力。评估的试剂盒代表了市售的 DNA 或总核酸提取方法的主要类型，包括 ChargeSwitch gDNA Mini Bacteria Kit、NucliSens Isolation Kit、Puregene Genomic DNA Purification Kit、QIAamp DNA Blood Mini Kit 和 UltraClean Microbial DNA Isolation Kit。使用八种炭疽芽孢杆菌强毒菌株的孢子进行提取方法。核酸提取物的活力测试表明，UltraClean 试剂盒在去除活炭疽芽孢杆菌孢子样本方面最为有效。 TaqMan 实时 PCR 分析表明，NucliSens、QIAamp 和 Ultraclean 试剂盒对孢子悬浮液的检测水平最高。对加标拭子和三种粉末类型的处理样品进行比较表明，使用 UltraClean 试剂盒提取 DNA 可获得最一致的阳性结果和最低的检测限。这项研究表明，不同的核酸提取方法（此处以各种商业提取试剂盒为代表）在灭活活炭疽芽孢杆菌孢子的能力以及 DNA 产量和纯度方面存在差异。此外，所使用的提取方法会影响炭疽芽孢杆菌实时 PCR 检测的灵敏度。由 Elsevier B.V. 出版
This study evaluated five commercial extraction kits for their ability to recover DNA from Bacillus anthracis spores and spiked environmental samples. The kits evaluated represent the major types of methodologies which are commercially available for DNA or total nucleic acid extraction, and included the ChargeSwitch gDNA Mini Bacteria Kit, NucliSens Isolation Kit, Puregene Genomic DNA Purification Kit, QIAamp DNA Blood Mini Kit, and the UltraClean Microbial DNA Isolation Kit. Extraction methods were performed using the spores of eight virulent strains of B. anthracis. Viability testing of nucleic acid extracts showed that the UltraClean kit was the most efficient at depleting samples of live B. anthracis spores. TaqMan real-time PCR analysis revealed that the NucliSens, QIAamp and Ultraclean kits yielded the best level of detection from spore suspensions. Comparisons of processed samples from spiked swabs and three powder types indicated that DNA extraction using the UltraClean kit resulted in the most consistently positive results and the lowest limit of detection. This study demonstrated that different nucleic extraction methodologies, represented here by various commercial extraction kits, differ in their ability to inactivate live B. anthracis spores as well as DNA yield and purity. In addition, the extraction method used can influence the sensitivity of real-time PCR assays for B. anthracis. Published by Elsevier B.V.