Open AccessThe development and use of Actiphage ® to detect viable mycobacteria from bovine tuberculosis and Johne’s disease‐infected animals
Author(s) -
Swift Benjamin M. C.,
Meade Nathan,
Barron Elsa Sandoval,
Bennett Malcolm,
Perehenic Tania,
Hughes Valerie,
Stevenson Karen,
Rees Catherine E. D.
Publication year - 2020
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.13518
Subject(s) - mycobacterium bovis , mycobacterium , paratuberculosis , biology , microbiology and biotechnology , tuberculosis , virology , mycobacterium tuberculosis , lysis , bacteria , immunology , medicine , genetics , pathology
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. paratuberculosi s 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.