Science Inventory

Evaluation of a Modified Rapid Viability-Polymerase Chain Reaction Method for Bacillus atrophaeus Spores in Water Matrices

Citation:

Bushon, R., A. Brady, C. Kephart, AND Vincent Gallardo. Evaluation of a Modified Rapid Viability-Polymerase Chain Reaction Method for Bacillus atrophaeus Spores in Water Matrices. JOURNAL OF MICROBIOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, 188:106293, (2021). https://doi.org/10.1016/j.mimet.2021.106293

Impact/Purpose:

The article describes a study where a modified rapid viability PCR (MRV-PCR) method was evaluated and compared against traditional culture method. The MRV-PCR wash shown to be a promising method for identifying viable Bacillus spores in small and large volumes of water samples and for producing results in less time than traditional culture methods.

Description:

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 techniques to provide results on whether viable organisms are present in less than 15 hours. 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 percent. The total time from the start of sample processing to the final sample result for the MRV-PCR method was decreased by approximately 2 hours because of the incorporation of a shorter, more efficient primary and secondary concentration step and a shorter DNA extraction technique. Overall, this study showed that MRV-PCR is a promising method for identifying viable Bacillus spores in small and large volumes of water samples and for producing results in less time than traditional culture methods.