[Rapid detection of mutations related to Mycobacterium leprae drug resistance by using Hp-rPCR (hairpin primer- real time PCR) method].

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Title[Rapid detection of mutations related to Mycobacterium leprae drug resistance by using Hp-rPCR (hairpin primer- real time PCR) method].
Publication TypeJournal Article
AuthorsKai M
Abbrev. JournalNihon Hansenbyo Gakkai Zasshi
JournalNihon Hansenbyō Gakkai zasshi = Japanese journal of leprosy : official organ of the Japanese Leprosy Association
Year of Publication2014
Volume83
Issue1
Pagination6-13
Publication Languagejpn
KeywordsAnti-Bacterial Agents, Base Sequence, Codon, Dapsone, DNA Primers, DNA, Bacterial, Double-Blind Method, Drug Resistance, Bacterial, Genes, Bacterial, Leprostatic Agents, Molecular Sequence Data, Mutation, Mycobacterium leprae, Quinolones, Real-Time Polymerase Chain Reaction, Rifampin
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%.

PubMed URL

http://www.ncbi.nlm.nih.gov/pubmed/25076760?dopt=Abstract

Link to full texthttps://www.jstage.jst.go.jp/article/hansen/83/1/83_6/_pdf

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