title: Use of a macrophage cell line for rapid detection of Mycobacterium bovis in diagnostic samples.
abstract: Mycobacterium bovis isolation on bacteriological media from suspected cases of bovine tuberculosis (TB) demands laborious and time-consuming procedures. Even polymerase chain reaction (PCR) and radiometric analyses are secondary procedures and not alternatives to bacteriological procedures. Therefore, there is a need to develop new techniques aimed at rapid M. bovis detection in diagnostic samples. The human macrophage cell line THP-1 was thus investigated in experiments of M. bovis propagation and isolation from reference lymph node suspensions. THP-1 cells were shown to support a high-titered propagation within 48h of minute amounts of both M. bovis BCG and fully pathogenic M. bovis strain 503. A semi-nested PCR for TB-complex-specific insertion sequence IS6110 revealed M. bovis infection in THP-1 cells. The same was true of a flow cytometry (FC) assay for expression of M. bovis chaperonin 10 in infected cells. The reduced time for isolation and identification of M. bovis (48-72h) and the consistency of the test results make the use of macrophage cell cultures attractive and cost-effective for veterinary laboratories involved in TB surveillance.
authors.affiliation: Department of Immunoprophylaxis and Animal Welfare, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, via A. Bianchi 9, 25124, Brescia, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Ritelli
authors.forename: M
authors.initials: M
authors.lastname: Amadori
authors.forename: S
authors.initials: S
authors.lastname: Tagliabue
authors.forename: M L
authors.initials: ML
authors.lastname: Pacciarini
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002417:Cattle; D005434:Flow Cytometry / Q000662:veterinary; D006801:Humans; D008198:Lymph Nodes / Q000382:microbiology; D008264:Macrophages / Q000382:microbiology*; D009163:Mycobacterium bovis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D014380:Tuberculosis, Bovine / Q000175:diagnosis* / Q000382:microbiology; D014407:Tumor Cells, Cultured