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[结核分枝杆菌菌株链霉素敏感性与rpsL突变之间的关系]

[Relationship between streptomycin susceptibility and rpsL mutations of Mycobacterium tuberculosis strains].

作者信息

Fukuda M, Koga H, Ohno H, Ogawa K, Yang B, Miyamoto J, Tomono K, Kohno S

机构信息

Second Department of Internal Medicine, Nagasaki University School of Medicine, Japan.

出版信息

Kekkaku. 1997 Sep;72(9):507-13.

PMID:9364810
Abstract

The relationship between streptomycin (SM) susceptibility and rpsL mutations of Mycobacterium tuberculosis strains was studied. Of 18 clinically isolated SM-resistant M.tuberculosis strains, mutation was suspected in 9 strains (50%) with SM MICs of > or = 256 micrograms/ml by PCR-single strand conformation polymorphism targeting rpsL gene. On the other hand, using PCR-direct sequence method, amino acid substitution caused by single nucleotide point mutation in rpsL gene was demonstrated in 11 out of 18 strains (61%). The same amino acid substitution at codon 43 (Lys-->Arg) was observed in all 11 strains with SM MICs of > or = 256 micrograms/ml. In addition, PCR products obtained from these 11 strains could not be cut by a restriction enzyme, Mbo II, while H37Rv strain and the other 32 strains with SM MICs of < 256 micrograms/ml were cut into 2 fragments. In conclusion, our results suggest that highly SM-resistant M.tuberculosis strains with MICs of > or = 256 micrograms/ml could be rapidly and easily detected by the restriction enzymatic method.

摘要

研究了结核分枝杆菌菌株的链霉素(SM)敏感性与rpsL基因突变之间的关系。在18株临床分离的对SM耐药的结核分枝杆菌菌株中,通过针对rpsL基因的PCR-单链构象多态性分析,怀疑9株(50%)菌株发生了突变,这些菌株的SM MICs≥256微克/毫升。另一方面,使用PCR直接测序法,在18株中的11株(61%)中证实了rpsL基因中的单核苷酸点突变导致氨基酸替代。在所有11株SM MICs≥256微克/毫升的菌株中均观察到密码子43处相同的氨基酸替代(赖氨酸→精氨酸)。此外,从这11株菌株获得的PCR产物不能被限制性内切酶Mbo II切割,而H37Rv菌株和其他32株SM MICs<256微克/毫升的菌株被切割成2个片段。总之,我们的结果表明,通过限制性酶切方法可以快速、简便地检测出MICs≥256微克/毫升的高度耐SM结核分枝杆菌菌株。

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