Evaluation of the Speed-oligo® Mycobacteria assay for identification of Mycobacterium spp. from fresh liquid and solid cultures of human clinical samples

Diagn Microbiol Infect Dis. 2010 Oct;68(2):123-31. doi: 10.1016/j.diagmicrobio.2010.06.006.

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Typing Techniques
  • Cross Reactions
  • DNA, Bacterial / analysis*
  • DNA, Ribosomal / genetics
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Mycobacterium / classification*
  • Mycobacterium / genetics
  • Mycobacterium / isolation & purification*
  • Mycobacterium Infections / diagnosis*
  • 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
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal, 16S / genetics
  • RNA, Ribosomal, 23S / genetics
  • Reagent Kits, Diagnostic
  • Spain
  • Species Specificity

Substances

  • DNA, Bacterial
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S
  • RNA, Ribosomal, 23S
  • Reagent Kits, Diagnostic