title: Routine use of PCR-reverse cross-blot hybridization assay for rapid identification of Mycobacterium species growing in liquid media.
abstract: A PCR-reverse cross-blot hybridization assay procedure that is able to rapidly identify 13 species of clinically relevant mycobacteria was evaluated for routine use in the identification of acid-fast isolates growing in BACTEC 460 TB (12B and 13A) and BACTEC 9000 MB (Myco/F) liquid media. Eight of the probes used were already described by Kox et al. (L. F. F. Kox et al., J. Clin. Microbiol. 33:3225-3233, 1995). In addition, we used six other probes specific for M. chelonae, M. malmoense or M. szulgai, M. genavense, M. gordonae, M. terrae, and M. marinum/M. ulcerans that we designed ourselves. This procedure allowed us to identify 459 mycobacterial species directly from broth cultures of 5,466 clinical samples collected over 1 year and processed with the radiometric or nonradiometric BACTEC system. Our results were in agreement with those obtained by conventional identification methods and also with those obtained by mycolic acid analysis by high-performance liquid chromatography. This assay seems to be a reliable procedure for the routine identification of mycobacteria, providing an accurate identification of mycobacterial isolates more rapidly than conventional tests, with remarkable implications for an efficacious specific antimycobacterial therapy.
authors.affiliation: Istituto di Microbiologia, Universitá Cattolica del Sacro Cuore, Rome, Italy. ibimb@rm.unieatt.it
authors.forename: M
authors.initials: M
authors.lastname: Sanguinetti
authors.forename: B
authors.initials: B
authors.lastname: Posteraro
authors.forename: F
authors.initials: F
authors.lastname: Ardito
authors.forename: S
authors.initials: S
authors.lastname: Zanetti
authors.forename: A
authors.initials: A
authors.lastname: Cingolani
authors.forename: L
authors.initials: L
authors.lastname: Sechi
authors.forename: A
authors.initials: A
authors.lastname: De Luca
authors.forename: L
authors.initials: L
authors.lastname: Ortona
authors.forename: G
authors.initials: G
authors.lastname: Fadda
mesh_terms: D002851:Chromatography, High Pressure Liquid; D015169:Colony Count, Microbial; D003470:Culture Media; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D009161:Mycobacterium / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D009171:Mycolic Acids / Q000032:analysis; D009170:Nontuberculous Mycobacteria / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D003470:Culture Media; D015342:DNA Probes; D004269:DNA, Bacterial; D009171:Mycolic Acids