A rapid method that provides information on the viability of organisms is needed to protect public health and ensure that remediation efforts following a release of a biological agent are effective. The rapid viability-polymerase chain reaction (RV-PCR) method combines broth culture and molecular methods to provide results on whether viable organisms are present in less than 15 h. In this study, a modified RV-PCR (mRV-PCR) method was compared to a membrane-filtration culture method for the detection of viable Bacillus spores in water matrices. Samples included small and large volumes of chlorine and non‑chlorine treated tap water. Large volume water samples (up to 100 L), were processed by ultrafiltration using a semi-automated waterborne pathogen concentrator, followed by centrifugation as a secondary concentration technique. The concentrated samples were analyzed by mRV-PCR and culture methods. The overall agreement between the mRV-PCR and culture methods when seed concentrations were greater than 10 spores per sample volume analyzed was 96%. The total time from the start of sample processing to the final sample result for the mRV-PCR method was decreased by approximately 2 h, in comparison to the previously published RV-PCR method because of the incorporation of shorter, more efficient primary and secondary concentration steps and a shorter DNA extraction technique. Overall, this study confirmed that RV-PCR is a promising approach for identifying viable Bacillus spores in small- and large-volume water samples and for producing results in less time than traditional culture methods.

需要一种提供有关生物体活力信息的快速方法，以保护公众健康并确保生物制剂释放后的补救措施有效。快速活力-聚合酶链反应 (RV-PCR) 方法结合了肉汤培养和分子方法，可在不到 15 小时内提供有关是否存在活菌的结果。在这项研究中，将改进的 RV-PCR (mRV-PCR) 方法与膜过滤培养方法用于检测活芽孢杆菌水基质中的孢子。样品包括少量和大量氯和未经氯处理的自来水。使用半自动水传播病原体浓缩器通过超滤处理大体积水样（高达 100 L），然后将离心作为二次浓缩技术。通过mRV-PCR和培养方法分析浓缩样品。当种子浓度大于每分析样品体积 10 个孢子时，mRV-PCR 和培养方法之间的总体一致性为 96%。与先前公布的 RV-PCR 方法相比，mRV-PCR 方法从样品处理开始到最终样品结果的总时间减少了大约 2 小时，因为结合了更短、更有效的初级和次级浓度步骤和更短的DNA提取技术。总体而言，这项研究证实，RV-PCR 是一种很有前途的方法，可用于鉴定小体积和大体积水样中的活芽孢杆菌孢子，并且比传统培养方法在更短的时间内产生结果。