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[Rapid detection of mutations related to Mycobacterium leprae drug resistance by using Hp-rPCR (hairpin primer- real time PCR) method].
- Source :
-
Nihon Hansenbyo Gakkai zasshi = Japanese journal of leprosy : official organ of the Japanese Leprosy Association [Nihon Hansenbyo Gakkai Zasshi] 2014 Mar; Vol. 83 (1), pp. 6-13. - Publication Year :
- 2014
-
Abstract
- Rapid and simple detection method of drug resistance bacteria is required. In the present study, Hp-rPCR (hairpin primer-real time PCR) was applied to Mycobacterium leprae genes to detect mutations. Target sites of the method were as follows: first base and second base on 53rd codon and second base on 55th codon infolP1 gene for dapsone resistance, first base on 441st codon and 451st codon and second base on 456th and 458th codon in rpoB gene for rifampicin resistance, and first base on 89th codon and second base on 91st codon in gyrA gene for quinolone resistance which were common mutation sites in clinical reports. The total number of the target sites was 9. Mycobacterium leprae, Thai-53, Zensho-2 and Zensho-4 were used as reference bacteria in the present study and clear, reliable results were obtained. Double-blind study was conducted using 15 samples. The number of target sites was calculated as 135 in total by 9 sites in 15 samples. There was only one misreading in the blind samples and the sensitivity was more than 99%.
- Subjects :
- Base Sequence
Codon genetics
DNA Primers genetics
DNA, Bacterial genetics
Dapsone pharmacology
Double-Blind Method
Genes, Bacterial genetics
Molecular Sequence Data
Quinolones pharmacology
Rifampin pharmacology
Anti-Bacterial Agents pharmacology
Drug Resistance, Bacterial drug effects
Leprostatic Agents pharmacology
Mutation genetics
Mycobacterium leprae drug effects
Mycobacterium leprae genetics
Real-Time Polymerase Chain Reaction methods
Subjects
Details
- Language :
- Japanese
- ISSN :
- 1342-3681
- Volume :
- 83
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nihon Hansenbyo Gakkai zasshi = Japanese journal of leprosy : official organ of the Japanese Leprosy Association
- Publication Type :
- Academic Journal
- Accession number :
- 25076760
- Full Text :
- https://doi.org/10.5025/hansen.83.6