Lawrence Livermore National Laboratory, 7000 East Avenue (L-452), Livermore, CA 94550, United States.
J Microbiol Methods. 2010 May;81(2):200-2. doi: 10.1016/j.mimet.2010.02.011. Epub 2010 Mar 1.
A comparison of Most-Probable-Number Rapid Viability (MPN RV) PCR and traditional culture methods for the quantification of Bacillus anthracis Sterne spores in macrofoam swabs from a multi-center validation study was performed. The purpose of the study was to compare environmental swab processing methods for recovery, detection, and quantification of viable B. anthracis spores from surfaces. Results show that spore numbers provided by the MPN RV-PCR method were typically within 1-log of the values from a plate count method for all three levels of spores tested (3.1x10(4), 400, and 40 spores sampled from surfaces with swabs) even in the presence of debris. The MPN method tended to overestimate the expected result, especially at lower spore levels. Blind negative samples were correctly identified using both methods showing a lack of cross contamination. In addition to detecting low levels of spores in environmental conditions, the MPN RV-PCR method is specific, and compatible with automated high-throughput sample processing and analysis protocols, enhancing its utility for characterization and clearance following a biothreat agent release.
进行了最可能数快速活力（MPN RV）PCR 与传统培养方法对炭疽杆菌 Sterne 孢子在多中心验证研究中的宏观泡沫拭子中的定量比较。研究的目的是比较环境拭子处理方法，以从表面回收、检测和定量有活力的炭疽杆菌孢子。结果表明，对于所有三种测试的孢子水平（从表面用拭子取样的 3.1x10(4)、400 和 40 个孢子），MPN RV-PCR 方法提供的孢子数量通常与平板计数方法提供的数值相差 1 个对数级，即使存在碎片。MPN 方法往往会高估预期结果，尤其是在较低的孢子水平下。两种方法均正确识别盲阴性样本，表明不存在交叉污染。除了在环境条件下检测低水平的孢子外，MPN RV-PCR 方法具有特异性，并且与自动化高通量样品处理和分析协议兼容，增强了其在生物威胁剂释放后的特征描述和清除方面的实用性。
