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The Journal of the Japanese Society for Clinical Microbiology |
Biblioraphy Information
ArticleTitle |
Species specificity evaluation of GENECUBE reagents detecting mycobacteria |
Language |
J |
AuthorList |
Yosuke Kawashima1), Kinuyo Chikamatsu2), Akio Sugiyama3), Hiromichi Suzuki4), Satoshi Mitarai2) |
Affiliation |
1) Biotechnology Research Laboratory, TOYOBO CO., LTD.
2) Department of Mycobacterium Reference and Research, the Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association
3) Diagnostic System Department, TOYOBO CO., LTD.
4) Department of Infectious Diseases, Faculty of Medicine, University of Tsukuba |
Publication |
J.J.C.M.: 31 (3), 161-169, 2021 |
Received |
March 2, 2021 |
Accepted |
April 19, 2021 |
Abstract |
We evaluated the limit of detection (LoD) and cross-reactivity of GENECUBE MTB (G-MTB), GENECUBE MAC (G-MAC) and GENECUBE MAI (G-MAI) in 168 standard strains of Mycobacterium species. All reagents detected the target species correctly, and the LoD was found to be between McFarland No. 1×10−5 and 10−6.
All non-target species tested by G-MTB were negative, although a peak in the melting curve analysis was detected for Mycobacterium haemophilum in the low-temperature range (57°C). In the G-MAC assay, 14 non-target species showed peaks (9: false positive, 5: low temperature). In the G-MAI (M. intracellulare detection reagent, MIN), 15 non-target species showed peaks (7: false positive, 8 low temperature). In G-MAI (M. avium detection reagent, MAV), 16 non-target species showed peaks (0: false positive, 16: low temperature). Mycobacterium genavense, a culturable but very slow-growing organism even in liquid culture medium and generally reported to be culture negative, showed a unique pattern (MAC: positive, MIN: positive, MAV: low temperature). Mycobacterium lentiflavum showed a similar pattern. The difference in the LoD of M. genavense and M. lentiflavum was one thousand-fold. Subsequently, we suggest that specimens that are strongly smear-positive, culture-negative, and show this unique pattern in GENCUBE indicate an infection by M. genavense.
In this study, the characteristic detection patterns were obtained from the analysis of various Mycobacterium species with GENECUBE. This suggests that there is the possibility of exhaustive discrimination of Mycobacterium species by combination of some of Mycobacterium detection reagents using the Quenching probe. |
Keywords |
Mycobacterium tuberculosis, Non-tuberculosis mycobacterium, Cross-reactivity, GENECUBE, QProbe |
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