Amie M.G. Brady
Christopher M. Kephart
Vicente Gallardo
Rebecca N. Bushon
2021
<div id="abstracts" class="Abstracts u-font-serif"><div id="ab0005" class="abstract author" lang="en"><div id="as0005"><p id="sp0060">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<span> </span><i>Bacillus</i><span> 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 </span><i>Bacillus</i><span> </span>spores in small- and large-volume water samples and for producing results in less time than traditional culture methods.</p></div></div></div>
application/pdf
10.1016/j.mimet.2021.106293
en
Elsevier
Evaluation of a modified rapid viability-polymerase chain reaction method for Bacillus atrophaeus spores in water matrices
article