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

Autor: Benjamin M. C. Swift, Nathan Meade, Elsa Sandoval Barron, Malcolm Bennett, Tania Perehenic, Valerie Hughes, Karen Stevenson, Catherine E. D. Rees
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: Microbial Biotechnology, Vol 13, Iss 3, Pp 738-746 (2020)
Druh dokumentu: article
ISSN: 1751-7915
79487491
DOI: 10.1111/1751-7915.13518
Popis: 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.
Databáze: Directory of Open Access Journals
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