The Use of Phage for Detection, Antibiotic Sensitivity Testing and Enumeration

Rees, Catherine; Botsaris, George

doi:10.5772/29734

Cited by 9 publications

(12 citation statements)

References 43 publications

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“… Number of cells added to each sample was determined using the phage amplification enumeration method described by Rees and Botsaris (). …”

Section: Resultsmentioning

confidence: 99%

“…Enumeration of mycobacterial cells present in laboratory cultures was performed using a modification of the phage amplification assay which included serial dilution of the samples prior to plating, and the number of viable mycobacteria cells detected was reported as pfu ml ‐1 . For detailed schematics, see Rees and Botsaris ().…”

Section: Methodsmentioning

confidence: 99%

“…Bacteriophage (phage) are viruses that are natural predators of bacteria, and phage‐based detection methods for mycobacteria were first described for the detection of M. tuberculosis in humans (Rees and Botsaris, ). One such assay has been termed phage amplification and results in the formation of plaques in a lawn of the fast‐growing non‐pathogen M. smegmatis (Stewart et al , ) .…”

Section: Introductionmentioning

confidence: 99%

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The development and use of Actiphage^® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

Swift

Meade

Barron

et al. 2019

Microbial Biotechnology

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Summary Here, we describe the development of a method that exploits bacteriophage D29 as a lysis agent for efficient DNA extraction from low numbers of mycobacterial cells. This method (Actiphage®) used in combination with PCR achieved rapid and sensitive (LOD ≤ 10 cell ml−1) detection and identification of viable, pathogenic mycobacteria in blood samples within 6 h. We demonstrate that mycobacteriophage D29 can be used to detect a range of mycobacteria from clinical blood samples including both Mycobacterium tuberculosis complex and Mycobacterium avium subsp. paratuberculosis without the need for culture and confirms our earlier observations that a low‐level bacteraemia is associated with these infections in cattle. In a study of M. bovis‐infected cattle (n = 41), the sensitivity of the Actiphage® method was 95 % (95 % CI; 0.84–0.99) and specificity was 100 % (95% CI; 0.92–1). We further used Actiphage® to demonstrate viable Mycobacterium avium subsp. paratuberculosis is present in the blood of Johne’s infected cattle. This method provides a revolutionary new tool for the study of infections caused by these difficult to grow pathogens.

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“… Number of cells added to each sample was determined using the phage amplification enumeration method described by Rees and Botsaris (). …”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The development and use of Actiphage^® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

Swift

Meade

Barron

et al. 2019

Microbial Biotechnology

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“…Standard FPTB assay positive and negative controls were used each time the assay was performed. Enumeration of MAP cells in inocula was determined using the modification of the FPTB assay as described by Rees and Botsaris (2012) which involves diluting samples until countable numbers of plaques are obtained (data reported as pfu ml − 1 ).…”

Section: Methodsmentioning

confidence: 99%

Development of a rapid phage-based method for the detection of viable Mycobacterium avium subsp. paratuberculosis in blood within 48 h

Swift

Denton

Mahendran

et al. 2013

Journal of Microbiological Methods

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The aim of this study was to develop a methodology to rapidly detect viable Mycobacterium avium subsp. paratuberculosis (MAP) in clinical blood samples. MAP cells spiked into commercially available blood were recovered using optimised peptide-mediated magnetic separation (PMMS) and detected using a phage-based method, and the identity of the cells detected confirmed using nested-PCR amplification of MAP signature sequences (IS900). The limit of detection was determined to be 10 MAP cells per ml of blood and was used to detect MAP present in clinical bovine blood samples. Using the PMMS-phage method there was no difference when detecting MAP from whole blood or from isolated buffy coat. MAP was detected in animals that were milk-ELISA positive (15 animals) by PMMS-phage and no MAP was detected in blood samples from an accredited Johne's disease free herd (5 animals). In a set of samples from one herd (10 animals) that came from animals with variable milk ELISA status, the PMMS-phage results agreed with the positive milk-ELISA results in all but one case. These results show that the PMMS-phage method can detect MAP present in naturally infected blood. Total assay time is 48 h and, unlike PCR-based detection tests, only viable cells are detected. A rapid method for detecting MAP in blood could further the understanding of disseminated infection in animals with Johne's disease.

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“…Detection and enumeration testing was carried out according to Rees and Botsaris (). Briefly, to perform the phage amplification assay, cell culture samples (200 μL) was mixed with 800‐μL Middlebrook 7H9 broth supplemented with ADC and CaCl 2 (2 mmol/L).…”

Section: Methodsmentioning

confidence: 99%

Intracellular delivery of nano‐formulated antituberculosis drugs enhances bactericidal activity

Donnellan

Stone

Johnston

et al. 2017

J of Interdiscip Nanomed

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Tuberculosis kills more people worldwide than any other infectious disease. Treatment requires multiple drug therapy administered over long periods (6-24 months). The emergence of multidrug-resistant strains is a major problem, and with few new drugs in the pipeline, a novel modus operandi is urgently required. Solid drug nanoparticles (SDNs), a new development in nanomedicine, offer a fresh therapeutic approach. Here, we show that SDNs are more effective (50-fold) at killing pathogenic mycobacteria than aqueous forms of the same drug and can target mycobacteria internalised by macrophages, where bacilli reside. We demonstrate synthesis of dual and triple drug loaded SDNs, facilitating combination tuberculosis therapy. Our results suggest that by employing SDNs of existing antibiotics, it may be possible to improve drug delivery and therefore reduce drug dosage to lessen side effects and fight drug resistance.

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The Use of Phage for Detection, Antibiotic Sensitivity Testing and Enumeration

Cited by 9 publications

References 43 publications

The development and use of Actiphage^® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

The development and use of Actiphage^® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

Development of a rapid phage-based method for the detection of viable Mycobacterium avium subsp. paratuberculosis in blood within 48 h

Intracellular delivery of nano‐formulated antituberculosis drugs enhances bactericidal activity

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The Use of Phage for Detection, Antibiotic Sensitivity Testing and Enumeration

Cited by 9 publications

References 43 publications

The development and use of Actiphage® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

The development and use of Actiphage® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

Development of a rapid phage-based method for the detection of viable Mycobacterium avium subsp. paratuberculosis in blood within 48 h

Intracellular delivery of nano‐formulated antituberculosis drugs enhances bactericidal activity

Contact Info

Product

Resources

About

The development and use of Actiphage^® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals

The development and use of Actiphage^® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals