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耐氟喹诺酮类药物大肠埃希氏菌基因
突变的MAMA-PCR检测
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陶文琴1,2，曾振灵1，陈杖榴1，刘健华1
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（1.华南农业大学 兽医学院 广东省兽药研制与安全评价重点实验室，广东 广州510642；
2.广州大学 生命科学学院，广东 广州510006）
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摘要：
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采用MAMA-PCR法对72株已知突变菌株的突变情况进行了检测。结果，有2种gyrA 83位突变：TCG→TTG和TCG缺失；4种gyrA 87位突变：GAC→AAC、GAC→GGC、GAC→TAC和GAC→CAC；1种parC 80位突变：AGC→ATC以及4种parC 84位突变：GAA→AAA、GAA→GCA、GAA→GGA和GAA→GTA。采用同样的方法，对38株未知突变菌株的检测结果显示，gyrA 83、gyrA 87、parC 80和parC 84位的突变检出的正确率分别为97.4%、947%、81.6%和94.7%。结果表明，MAMA-PCR有望成为监测耐氟喹诺酮类药物大肠埃希氏菌的分子生物学常规方法。 |
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关键词：
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大肠埃希氏菌；氟喹诺酮类；耐药性；突变检测；错配扩增突变分析PCR法 |
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中图分类号：S 852.612 文献标识码：
A 文章编号：1673-4696(2006)07-0534-04 |
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MAMA-PCR detection of gyrA and parC gene
mutations in fluoroquinolone-resistant Escherichia coli |
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TAO Wen-qin1,2，ZENG Zhen-ling1，CHEN
Zhang-liu1，LIU Jian-hua1 |
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（1.Guangdong Provincial Key Laboratory for Veterinary Pharmaceutics Development and Safety
Evaluation/College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China；
2.School of Life Science，Guangzhou University, Guangzhou 510006, China） |
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Abstract: The gyrA and parC gene mutations in 72 known-mutant strains of fluoroquinolone-resistant Escherichia coli isolated from animals，humans and environment were detected by mismatch amplification mutation assay PCR(MAMA-PCR). The following mutations were found in the 72 strains: TCG→TTG or deletion in gyrA gene 83rd site, GAC→AAC，GGC，TAC or CAC in gyrA gene 87th site, AGC→ATC in parC gene 80th site, and GAA→AAA，GCA，GGA or GTA in parC gene 84th site. The MAMA-PCR detection of 38 unknown-mutant strains of fluoroquinolone-resistant E.coli showed that correct ratios of mutation detection of gyrA gene 83rd and 87th sites, and parC gene 80th and 84th sites were 974%,94.7%,81.6% and 94.7%, respectively. It was concluded that MAMA-PCR may provide a molecular tool for the routine detection of gyrA and parC genes mutations in E.coli. |
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Key words: Escherichia coli；fluoroquinolone；resistance；mutation detection；MAMA-PCR |
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