title: Improved detection of Mycobacterium bovis infection in bovine lymph node tissue using immunomagnetic separation (IMS)-based methods.
abstract: Immunomagnetic separation (IMS) can selectively isolate and concentrate Mycobacterium bovis cells from lymph node tissue to facilitate subsequent detection by PCR (IMS-PCR) or culture (IMS-MGIT). This study describes application of these novel IMS-based methods to test for M. bovis in a survey of 280 bovine lymph nodes (206 visibly lesioned (VL), 74 non-visibly lesioned (NVL)) collected at slaughter as part of the Northern Ireland bovine TB eradication programme. Their performance was evaluated relative to culture. Overall, 174 (62.1%) lymph node samples tested positive by culture, 162 (57.8%) by IMS-PCR (targeting IS6110), and 191 (68.2%) by IMS-MGIT culture. Twelve (6.9%) of the 174 culture positive lymph node samples were not detected by either of the IMS-based methods. However, an additional 79 M. bovis positive lymph node samples (27 (13.1%) VL and 52 (70.3%) NVL) were detected by the IMS-based methods and not by culture. When low numbers of viable M. bovis are present in lymph nodes (e.g. in NVLs of skin test reactor cattle) decontamination prior to culture may adversely affect viability, leading to false negative culture results. In contrast, IMS specifically captures whole M. bovis cells (live, dead or potentially dormant) which are not subject to any deleterious treatment before detection by PCR or MGIT culture. During this study only 2.7% of NVL lymph nodes tested culture positive, whereas 70.3% of the same samples tested M. bovis positive by the IMS-based tests. Results clearly demonstrate that not only are the IMS-based methods more rapid but they have greater detection sensitivity than the culture approach currently used for the detection of M. bovis infection in cattle. Adoption of the IMS-based methods for lymph node testing would have the potential to improve M. bovis detection in clinical samples.
authors.affiliation: Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, Northern Ireland, United Kingdom.
authors.forename: Linda D
authors.initials: LD
authors.lastname: Stewart
authors.forename: James
authors.initials: J
authors.lastname: McNair
authors.forename: Lyanne
authors.initials: L
authors.lastname: McCallan
authors.forename: Alan
authors.initials: A
authors.lastname: Gordon
authors.forename: Irene R
authors.initials: IR
authors.lastname: Grant
mesh_terms: D000818:Animals; D002417:Cattle; D003470:Culture Media; D018189:Immunomagnetic Separation / Q000379:methods* / Q000662:veterinary*; D008198:Lymph Nodes / Q000382:microbiology; D009163:Mycobacterium bovis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014380:Tuberculosis, Bovine / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D003470:Culture Media