ArticleApplied and environmental microbiology2009
Optimization of a phage amplification assay to permit accurate enumeration of viable Mycobacterium avium subsp. paratuberculosis cells.
Article in Applied and environmental microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 53 citations in OpenAlex.
- Optimizing Bacteriophage Screening and Isolation Methods for Microbial Samples Derived from Different Body Sites of Cattle.Microorganisms · 2026Article
- Reduction of Salmonella gallinarum in poultry with phage-based interventions.Scientific reports · 2025Article
- Optimizing bacteriophage screening and isolation methods for microbial samples derived from different body sites of cattle.bioRxiv : the preprint server for biology · 2025Article
- Advances in detection methods for viable Salmonella spp.: current applications and challenges.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2023Review
- Utilisation of Actiphage in combination with ISJournal of veterinary research · 2023Article
- Mycobacteriophages in diagnosis and alternative treatment of mycobacterial infections.Frontiers in microbiology · 2023Review
- A Review on Mycobacteriophages: From Classification to Applications.Pathogens (Basel, Switzerland) · 2022Review
- MAP, Johne's disease and the microbiome; current knowledge and future considerations.Animal microbiome · 2021Review
- Article
- Phage Amplification Assay for Detection of Mycobacterial Infection: A Review.Microorganisms · 2021Review
- Application of the Actiphage® Assay to Detect ViableFrontiers in veterinary science · 2021Article
- Bacteriophage-Based Methods for Detection of ViableFrontiers in veterinary science · 2021Review
- Presence of Infection byMicroorganisms · 2020Article
- A novel one-day phage-based test for rapid detection and enumeration of viable Mycobacterium avium subsp. paratuberculosis in cows' milk.Applied microbiology and biotechnology · 2020Article
- Methods for detection of viable foodborne pathogens: current state-of-art and future prospects.Applied microbiology and biotechnology · 2020Review
- Crohn's disease: failure of a proprietary fluorescent in situ hybridization assay to detect M. avium subspecies paratuberculosis in archived frozen intestine from patients with Crohn's disease.BMC research notes · 2020Article
- Estimation of Performance Characteristics of Analytical Methods forFrontiers in microbiology · 2019Article
- In vitro inactivation of Mycobacterium avium subsp. paratuberculosis (MAP) by use of copper ions.BMC microbiology · 2018Article
- Failure to detect M. avium subspecies paratuberculosis in Johne's disease using a proprietary fluorescent in situ hybridization assay.BMC research notes · 2018Article
- Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A commercially available phage amplification assay, FASTPlaqueTB (Biotec Laboratories, Ipswich, United Kingdom), when used according to the manufacturer's instructions, does not permit accurate enumeration of Mycobacterium avium subsp. paratuberculosis. The aim of this study was to optimize the phage amplification assay conditions to permit accurate quantification of viable M. avium subsp. paratuberculosis cells. The burst time for M. avium subsp. paratuberculosis was initially determined to inform decisions about optimal incubation time before plating, and then other test parameters were altered to evaluate how the correlation between plaque and colony counts was affected. The D29 mycobacteriophage replicates more slowly in M. avium subsp. paratuberculosis than in Mycobacterium smegmatis (used to optimize the commercial test originally), and the mean burst time for four M. avium subsp. paratuberulosis strains was 210 +/- 36.8 min at 37 degrees C compared to 63 +/- 17.5 min for M. smegmatis mc(2) 155. To achieve 100% correlation between plaque and colony counts, the optimized phage assay includes the following: (i) resuspension of the samples to be tested in Middlebrook 7H9 broth containing 10% oleic acid-albumin-dextrose-catalase and 2 mM calcium chloride, followed by overnight incubation at 37 degrees C before performance of the phage assay; (ii) a 2-h incubation of the sample with D29 mycobacteriophage before viricide treatment; and (iii) a further 90-min incubation after viricide treatment and neutralization up to the burst time (total incubation time, 210 min) before plating with M. smegmatis mc(2) 155 in 7H9 agar. The optimized phage amplification assay was able to detect 1 to 10 CFU/ml of M. avium subsp. paratuberculosis in spiked milk or broth within 48 h, as demonstrated by the results of several blind trials.
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