鸡源性大肠埃希氏菌对氟喹诺酮类药物的耐药性研究
发布时间：2018-11-10 11:11
【摘要】： 目的利用分子生物学的方法检测太原地区鸡源性大肠埃希氏菌对氟喹诺酮类药物的耐药情况,旨在探讨喹诺酮类药物耐药决定区(QRDR)的促旋酶、拓扑异构酶以及孔道蛋白OmpC、OmpF基因变异情况。 方法采集山西省畜牧研究所种鸡场30株鸡源性大肠杆菌,用纸片法筛选出12株耐氟喹诺酮类药菌的大肠杆菌,分别用gyrA、gyrB、parC、parE、ompC、ompF基因相应的引物对12株耐氟喹诺酮类药菌的大肠杆菌进行PCR扩增,并选择其中4株有代表性菌株进行基因序列的测定;用SDS-PAGE电泳对12株耐药菌的外膜蛋白进行分析;用脉冲场凝胶电泳对12株耐药菌进行了流行情况分析。 结果所有检测株中,其中12株菌对环丙沙星耐药,占所采集标本的40%;PCR实验gyrA、gyrB、parC、parE、ompC、ompF都有清晰的目的条带;SDS-PAGE电泳结果显示鸡4、鸡5号菌株外膜蛋白OmpC的表达缺失;脉冲场凝胶电泳结果分析显示,存在小范围的流行,爆发流行的几率较小;测序结果显示,送检的测序株都存在GyrA在83或87位氨基酸的变异(Ser-83→Leu;Asp-87→Asn);鸡5号菌株ParC在80位由由丝氨酸变位亮氨酸(Ser-80→Leu),鸡6号菌株ParC在第84位由谷氨酸变为赖氨酸(Glu-84→Lys);鸡5号菌株ParE在89位丝氨酸变为丙氨酸(Ser-89→Ala)。 结论山西省畜牧研究所养鸡场的大肠杆菌对氟喹诺酮耐药的主要机制是由于促旋酶在第83氨基酸、87位氨基酸发生一点或同时发生变异造成的;另外,部分菌株伴随parC和ParE的突变可能是造成氟喹诺酮类药物高水平耐药的主要因素。部分菌株存在外膜蛋白OmpC的缺失,但是OmpC的缺失不是造成以上菌株对氟喹诺酮类抗菌素耐药的主要原因。
[Abstract]:Objective to detect the resistance of Escherichia coli from chicken to fluoroquinolones in Taiyuan area by molecular biology, and to investigate the protopolase, topoisomerase and pore protein OmpC, of quinolone drug resistance determining region (QRDR). Variation of OmpF gene. Methods 30 strains of Escherichia coli were collected from chicken farm of Shanxi Animal Husbandry Institute. Twelve strains of Escherichia coli resistant to fluoroquinolones were screened by disk method. GyrA,gyrB,parC,parE,ompC, was used respectively. The corresponding primers of ompF gene were used to amplify 12 strains of Escherichia coli resistant to fluoroquinolones and 4 representative strains were selected for gene sequencing. The outer membrane proteins of 12 strains of drug-resistant bacteria were analyzed by SDS-PAGE electrophoresis, and the epidemic situation of 12 strains of resistant bacteria was analyzed by pulsed field gel electrophoresis. Results among all the tested strains, 12 strains were resistant to ciprofloxacin, accounting for 40% of the collected samples. The results of SDS-PAGE electrophoresis showed that the expression of outer membrane protein (OmpC) of chicken strain 4 and chicken 5 was absent, the results of pulse field gel electrophoresis showed that there was a small epidemic, and the probability of outbreak was lower. The sequencing results showed that the Ser-83 Leu;Asp-87 Asn); existed in the GyrA at 83 or 87 amino acids in all the tested strains. Chicken 5 strain ParC changed from Ser-80 Leu), chicken 6 ParC to Glu-84 Lys); at 84th position from Ser-80 serine to Glu-84 Lys); The serine of chicken 5 strain ParE was transformed into Ser-89 Ala). At 89 position. Conclusion the main mechanism of Escherichia coli resistance to fluoroquinolone in chicken farm of Shanxi Animal Husbandry Research Institute is due to the mutation of a little or both amino acids at 83 amino acid and 87 amino acids. In addition, the mutation of parC and ParE in some strains may be the main factor of high level resistance of fluoroquinolones. The absence of outer membrane protein (OmpC) was found in some strains, but the deletion of OmpC was not the main reason for the resistance to fluoroquinolones.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378
本文编号：2322309
[Abstract]:Objective to detect the resistance of Escherichia coli from chicken to fluoroquinolones in Taiyuan area by molecular biology, and to investigate the protopolase, topoisomerase and pore protein OmpC, of quinolone drug resistance determining region (QRDR). Variation of OmpF gene. Methods 30 strains of Escherichia coli were collected from chicken farm of Shanxi Animal Husbandry Institute. Twelve strains of Escherichia coli resistant to fluoroquinolones were screened by disk method. GyrA,gyrB,parC,parE,ompC, was used respectively. The corresponding primers of ompF gene were used to amplify 12 strains of Escherichia coli resistant to fluoroquinolones and 4 representative strains were selected for gene sequencing. The outer membrane proteins of 12 strains of drug-resistant bacteria were analyzed by SDS-PAGE electrophoresis, and the epidemic situation of 12 strains of resistant bacteria was analyzed by pulsed field gel electrophoresis. Results among all the tested strains, 12 strains were resistant to ciprofloxacin, accounting for 40% of the collected samples. The results of SDS-PAGE electrophoresis showed that the expression of outer membrane protein (OmpC) of chicken strain 4 and chicken 5 was absent, the results of pulse field gel electrophoresis showed that there was a small epidemic, and the probability of outbreak was lower. The sequencing results showed that the Ser-83 Leu;Asp-87 Asn); existed in the GyrA at 83 or 87 amino acids in all the tested strains. Chicken 5 strain ParC changed from Ser-80 Leu), chicken 6 ParC to Glu-84 Lys); at 84th position from Ser-80 serine to Glu-84 Lys); The serine of chicken 5 strain ParE was transformed into Ser-89 Ala). At 89 position. Conclusion the main mechanism of Escherichia coli resistance to fluoroquinolone in chicken farm of Shanxi Animal Husbandry Research Institute is due to the mutation of a little or both amino acids at 83 amino acid and 87 amino acids. In addition, the mutation of parC and ParE in some strains may be the main factor of high level resistance of fluoroquinolones. The absence of outer membrane protein (OmpC) was found in some strains, but the deletion of OmpC was not the main reason for the resistance to fluoroquinolones.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378
【参考文献】
相关期刊论文 前10条
1 陈晓燕;107例老年患者肺部感染临床分析[J];北京医学;2001年04期
2 徐成刚,任涛,廖明,罗开健,辛朝安;兽用抗生素对公共卫生的影响[J];动物医学进展;2002年06期
3 吴聪明,陈杖榴;细菌耐药性扩散的机制[J];动物医学进展;2003年04期
4 张致平;喹诺酮类抗菌药研究的新进展[J];国外医药(抗生素分册);2001年06期
5 刘高生,卢升云;养殖业滥用抗生素贻害无穷[J];黑龙江畜牧兽医;2004年12期
6 顾宝柯,许学斌,金汇明,胡培玉,席曼芳;上海地区家畜、家禽及肉制品大肠杆菌O157∶H7监测[J];疾病监测;2003年01期
7 王辉,陈民钧;环丙沙星耐药大肠埃希菌30株的遗传特征[J];中国抗感染化疗杂志;2002年02期
8 李方去,李向阳,杨锦红;头孢西丁对三代头孢菌素耐药肠杆菌科某些菌种的作用[J];浙江临床医学;2002年03期
9 倪语星,韩立中;分子生物学方法在细菌耐药性检测方面的应用[J];上海医学检验杂志;2001年06期
10 李凯年;抗生素饲料添加剂的作用与发展趋势[J];饲料广角;2005年13期
，本文编号：2322309
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2322309.html
最近更新
教材专著
