Evaluation of the GenoType Mycobacterium Assay for Identification of Mycobacterial Species from Cultures

Abstract

A new commercially available DNA strip assay (GenoType Mycobacterium CM/AS; Hain Lifescience, Nehren, Germany) was evaluated for the ability to differentiate mycobacterial species. The test is based on a PCR technique targeting a 23S rRNA gene region, followed by reverse hybridization and line probe technology. The GenoType CM is capable of identifying 23, the GenoType AS a further 14, species either alone or in combination with one or more species. Both tests were evaluated with 156 mycobacterial strains composed of 61 validly published species including different subspecies, 6 not validly published species, and 3 strains other than mycobacterial species. All strains were precharacterized by sequencing of the 5 region of the 16S rRNA gene and biochemical tests. In total, results for 151 strains were interpretable. Concordant results were obtained for 137 (92.6%) of 148 mycobacterial strains with the CM assay and 133 (89.9%) of 148 mycobacterial strains with the AS assay, and all three non-Mycobacterium species were identified.

Clinical Laboratory Diagnostics for Mycobacterium tuberculosis

“…The overall sensitivity and specificity for species identification was 97.9% and 92.4% for the CM kit and 99.3% and 99.4% for the AS kit with all members of the Mtb complex correctly identified as Mtb complex by the CM kit and Mtb species by the AS kit. Similar performance were established in other independent studies evaluating the GenoType CM and AS kits, with the sensitivity and specificity for Mtb complex members approaching 100% when compared to 16S rRNA sequencing and biochemical testing (Makinen et al, 2006;Richter et al, 2006).…”

“…One kit, the Genotype Mycobacterium CM (Common Mycobacteria) was designed to detect the most frequently isolated mycobacteria species in clinical laboratories while the other kit, the Genotype Mycobacterium AS (Additional Species), was designed for the detection of less frequently encountered mycobacteria species (Makinen et al, 2006;Richter et al, 2006;Russo et al, 2006). Russo and colleagues tested 197 isolates including genera other than mycobacteria previously identified by conventional biochemical tests, HPLC, Inno-Lipa and 16S rRNA sequencing (Russo et al, 2006).…”

Evaluation of the GenoType® NTM DR for Subspecies Identification and Determination of Drug Resistance in Clinical M. abscessus Isolates

“…DNA was extracted as described elsewhere [16]. A senior scientist interpreted the results as per package insert.…”

Genetic Diversity and Population Structure of Mycobacterium marinum: New Insights into Host and Environmental Specificities

“…All 89 cultures (human and fish isolates) were positive by Ziehl-Neelsen staining. Based on conventional biochemical methods and the commercial multiplex line-probe assay GenoType Mycobacterium AS/CM (Hain Lifescience GmbH, Nehren, Germany), all 89 isolates were assigned to the species M. marinum (17,23).…”