Cloning and nucleotide sequence of the drug resistant genes and its application to rapid identification.

耐药基因的克隆、核苷酸序列及其快速鉴定应用。

• 批准号: 05670378
• 负责人: KATSUKAWA Chihiro
• 金额: $ 1.47万
• 依托单位: Osaka Prefectural Institute for Public Health
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670378/
• 关键词: Mycobacterium tuberculosis; Rifampicin; Streptomycin; Drug resistance; rpoB gene; rpsL gene; rrs gone; Mutation; 耐性; 30Sリボソーム; rrsL遺伝子; -

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.

结核分枝杆菌临床分离株对利福平和链霉素耐药的分子机制已被研究。我们用PCR技术测定了RNA聚合酶β亚基（rpoB）、核糖体S12蛋白（rpsL）和16 SrRNA（rrs）的基因序列。主要结论如下：1、对64株日本临床分离株的rpoB基因进行了全序列测定。遗传学方法可检出90%以上的RFP耐药株。2、采用PCR扩增173株日本临床分离株rpsL基因，并经Mbo Ⅱ酶切鉴定，将其分为两组。60.0%的SM耐药株（200 μ g/ml）不携带Mbo Ⅱ位点，而SM耐药株（20 μ g/ml）和SM敏感株则携带Mbo Ⅱ位点。对47株日本临床分离株的rrs基因进行了PCR扩增和序列测定。35.1%的SM耐药菌株携带点突变或移码插入。遗传学方法可检测出75%以上的SM抗性菌株。

项目成果

Yasuhiko Suzuki, Yue Ping Yin, Chihiro Katsukawa, Masanao Makino: "Overproduction of Recombinant 85 Complex of M.leprae in E.coli and Their Application for Serodiagnosis of Leprosy" Proceedings of 29th Joint Research Conference on Tuberculosis and Leprosy

Yasuhiko Suzuki、Yue Ping Yin、Chihiro Katsukawa、Masanao Makino：“大肠杆菌中麻风杆菌重组85复合物的过量生产及其在麻风病血清诊断中的应用”第29届结核病与麻风病联合研究会议论文集

Y. Suzuki, C. Katsukawa, K. Inoue, M. Tasaka, Y. P. Yin, N Ueba and M. Makino: "Mutations in rpo B Gene of Rifampicin Resistant Clinical Isolates of Mycobacterium" 感染症学雑誌. 69. 413-419 (1995)

Y. Suzuki、C. Katsukawa、K. Inoue、M. Tasaka、Y. P. Yin、N Ueba 和 M. Makino：“利福平耐药临床分离株的 rpo B 基因突变”《传染病杂志》69。413-419。 (1995)

Chihiro Katsukawa: "Cloning and Nucleotide Sequence of Type 3 M protein Gene(emm3)Consisting of an N-terminal Variable Portion and C-terminal Conserved C Repeat Regions" 感染症学雑誌. 68. 698-705 (1994)

Chihiro Katsukawa：“由 N 末端可变部分和 C 末端保守 C 重复区域组成的 3 型 M 蛋白基因 (emm3) 的克隆和核苷酸序列”传染病杂志 68. 698-705 (1994)。

Yasuhiko Suzuki, Yue Ping Yin, Chihiro Katsukawa and Masanao Makino: "Application of Recombinant 85 Complex of M. leprae for serodiagnosis of Leprosy" Proceedings of Thirtieth Joint Conference on Tuberculosis and Leprosy. U. S. Japan Cooperative Medical S

Yasuhiko Suzuki、Yue Ping Yin、Chihiro Katsukawa 和 Masanao Makino：“麻风分枝杆菌重组 85 复合物在麻风病血清诊断中的应用”第三十届结核病和麻风病联席会议论文集。

Chihiro Katsukawa, Aki Tamaru, Yasuhiko Suzuki, Masanao Makino: "Mutations in rpsL and rrs Gene of Streptomycin Resistant Isolates of Mycobacterium tuberculosis in Japan" The Journal of the Japanese Association for Infectious Diseases.

Chihiro Katsukawa、Aki Tamaru、Yasuhiko Suzuki、Masanao Makino：“日本结核分枝杆菌链霉素抗性分离株的 rpsL 和 rrs 基因突变”日本传染病协会杂志。