1995 Fiscal Year Final Research Report Summary
Cloning and nucleotide sequence of the drug resistant genes and its application to rapid identification.
| Project/Area Number |
05670378
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Public health/Health science
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| Research Institution | Osaka Prefectural Institute for Public Health |
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Principal Investigator |
KATSUKAWA Chihiro Osaka Prefectural Institute for Public Health Dept. Microbiol., Cheaf resercher, 微生物課, 主任研究員 (20183725)
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| Co-Investigator(Kenkyū-buntansha) |
MAKINO Masanao Osaka Prefectural Institute for Public Health Dept. Microbiol., Head of Dept., 微生物課, 微生物課長 (00116097)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Mycobacterium tuberculosis / Rifampicin / Streptomycin / Drug resistance / rpoB gene / rpsL gene / rrs gone / Mutation |
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| Research Abstract |
The molecular mechanisms of resistance to rifampicin and streptomycin in Mycobacterium tuberculosis clinical isolates have been examined. We have sequenced genes encoding the beta subunit of RNA polymerase (rpoB), the ribosomal S12 protein (rpsL) and 16SrRNA (rrs) by using PCR technique. Following conclusions have been obtained.
1, The rpoB genes of 64 Japanese clinical isolates have been sequenced. More than 90% of RFP resistant strain can be detected by genetical method.
2, The rpsL gene of 173 Japanese clinical isolates have been amplified by PCR and classified by Mbo II restriction digestion into two groups. 60.0% of SM-resistant (200mug/ml) isolates did not carry Mbo II site, whereas SM-resistant (20mug/ml) isolates and SM-sensitive isolates carried it. Another mutation was found on the aa88 by sequencing. The rrs gene of 47 Japanese clinical isolates have been amplified by PCR and sequenced. 35.1% of SM-resistant isolates carried a point mutation or frame shift insertion. More than 75% of SM resistant strain can be detected by genetical method.
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