title: Evaluation of the Speed-oligo® Mycobacteria assay for identification of Mycobacterium spp. from fresh liquid and solid cultures of human clinical samples.
abstract: We evaluated the ability of a novel DNA strip assay (Speed-oligo® Mycobacteria) to differentiate mycobacterial species. It is based on polymerase chain reaction targeting 16S rRNA and 16S-23S rRNA regions and double-reverse hybridization on a dipstick using probes bound to colloidal gold and to the membrane. We blindly tested its capacity to identify 182 acid-fast bacilli grown on fresh liquid (BacT/Alert, MGIT) and solid (Lowenstein-Jensen) cultures (from Spanish mycobacteriology laboratories), previously identified by means of Genotype(®) Mycob.CM/AS or Gen-Probe(®) AccuprobeMTC, and 11 collection strains of mycobacteria-related organisms. Discrepancies were resolved by 16S rRNA sequencing. Results were interpreted by identification of 7 specific bands for the following: Mycobacterium sp., M. fortuitum, Mycobacterium avium-intracellulare complex, Mycobacterium tuberculosis complex, Mycobacterium kansasii, M. gordonae, and M. abscessus-chelonae complex. No cross-reactivity was observed with any mycobacteria-related organism. Concordant results were obtained for 177/182 bacilli (97.2%). There was only 1 major discrepancy, misidentification of Mycobacterium marinum as M. kansasii, verified by sequencing.
authors.affiliation: Department of Microbiology, E.P.H. Costa del Sol, Marbella (Málaga), Spain.
authors.forename: Natalia Montiel
authors.initials: NM
authors.lastname: Quezel-Guerraz
authors.forename: Mercedes Marín
authors.initials: MM
authors.lastname: Arriaza
authors.forename: José Antonio Carrillo
authors.initials: JA
authors.lastname: Avila
authors.forename: Waldo E
authors.initials: WE
authors.lastname: Sánchez-Yebra Romera
authors.forename: Miguel J
authors.initials: MJ
authors.lastname: Martínez-Lirola
mesh_terms: D015373:Bacterial Typing Techniques; D003429:Cross Reactions; D004269:DNA, Bacterial / Q000032:analysis*; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D009164:Mycobacterium Infections / Q000175:diagnosis*; D009162:Mycobacterium avium / Q000302:isolation & purification; D015269:Mycobacterium avium Complex / Q000302:isolation & purification; D019909:Mycobacterium kansasii / Q000302:isolation & purification; D019910:Mycobacterium marinum / Q000302:isolation & purification; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification; D009170:Nontuberculous Mycobacteria / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D013030:Spain; D013045:Species Specificity
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D011933:Reagent Kits, Diagnostic