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Evaluation of a modified rapid viability-polymerase chain reaction method for Bacillus atrophaeus spores in water matrices
- Source :
- J Microbiol Methods
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- 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.
- Subjects :
- DNA, Bacterial
Microbiology (medical)
Sample (material)
Ultrafiltration
chemistry.chemical_element
Bacillus
Polymerase Chain Reaction
Sensitivity and Specificity
Microbiology
Article
Tap water
Chlorine
Centrifugation
Molecular Biology
Spores, Bacterial
Bacteriological Techniques
Microbial Viability
Chromatography
biology
Water
biology.organism_classification
DNA extraction
Spore
chemistry
Bacillus atrophaeus
Water Microbiology
Subjects
Details
- ISSN :
- 01677012
- Volume :
- 188
- Database :
- OpenAIRE
- Journal :
- Journal of Microbiological Methods
- Accession number :
- edsair.doi.dedup.....084883672d19de8a448bc2fbefd46364
- Full Text :
- https://doi.org/10.1016/j.mimet.2021.106293