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Diagn Microbiol Infect Dis ; 68(2): 123-31, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20846584

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.


Subject(s)
DNA, Bacterial/analysis , Molecular Diagnostic Techniques/methods , Mycobacterium Infections/diagnosis , Mycobacterium/classification , Mycobacterium/isolation & purification , Polymerase Chain Reaction/methods , Bacterial Typing Techniques , Cross Reactions , DNA, Ribosomal/genetics , Humans , Mycobacterium/genetics , Mycobacterium avium/isolation & purification , Mycobacterium avium Complex/isolation & purification , Mycobacterium kansasii/isolation & purification , Mycobacterium marinum/isolation & purification , Mycobacterium tuberculosis/isolation & purification , Nontuberculous Mycobacteria/isolation & purification , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 23S/genetics , Reagent Kits, Diagnostic , Spain , Species Specificity
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