The GenoType® Mycobacterium CM assay (Hain Lifescience, Nehren Germany) was used to analyze NTM isolates at the species level, and the procedure described in the manual enclosed in the kit was followed to conduct the test. The assay involved DNA amplification targeting the 23S rRNA gene region, as recommended. Followed by the reverse hybridization to specific oligonucleotide probes immobilized on membrane strips, which was conducted on a shaking TwinCubator (Hain). The final result was interpreted based on the presence and absence of bands, and compared with the evaluation sheet provided with the kit. M. tuberculosis H37Rv, M. fortuitum and M. abscessus were used as appositive control while H2O Qiagen as a negative control.
Genotype mycobacterium cm assay
Manufactured by Hain Lifescience
Sourced in Germany
The GenoType® Mycobacterium CM assay is a molecular diagnostic test designed to identify and differentiate between the most common nontuberculous mycobacteria (NTM) species. The assay utilizes DNA-strip technology to provide reliable and reproducible results for the identification of Mycobacterium species.
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2 protocols using genotype mycobacterium cm assay
1
Identification of Nontuberculous Mycobacteria
2
GenoType Mycobacterium CM Assay Protocol
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GenoType Mycobacterium CM assay (Hain Life Science, Nehren, Germany) was performed to identify mycobacteria species, according to the manufacturer’s instructions. The assay was run on a GT-Blot 48 (Hain Life Science, Nehren, Germany). The volume of the PCR mixture was 50 μL, comprised of 35 μL PNM, 5 μL 10x PCR buffer for HotStarTaq DNA Polymerase (Qiagen, Germany), 2 μL 25 mM MgCl2 solution, 1 U HotStarTaq DNA Polymerase, 3 μL deionized water, and 5 μL DNA solution. PCR was performed by denaturation steps at 95℃ for 15 min, 10 cycles of 95℃ for 30s and 58℃ for 2 min, 20 cycles of 95℃ for 25s, 53℃ for 40s, and 70℃ for 40s, followed by an extension at 70℃ for 8 min. After PCR completion, the results were automatically analyzed by the integrated software.
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