福建省莆田市鸭疫里默氏杆菌的喹诺酮类耐药基因检测与QRDR基因突变分析  

Detection of Quinolone Resistance Genes and QRDR Mutation in Riemerella anatipestifer Found in Putian, Fujian

作　　者：林彬彬[1] 林志敏[1] 谢碧林 翁汉东[1] 王秀祯 程龙飞[2] 傅光华[2] 刘荣昌[2] LIN Binbin;LIN Zhimin;XIE Bilin;WENG Handong;WANG Xiuzhen;CHENG Longfei;FU Guanghua;LIU Rongchang(Putian Institute of Agricultural Sciences,Putian,Fujian 351144,China;Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350013,China)

机构地区：[1]福建省莆田市农业科学研究所,福建莆田351144 [2]福建省农业科学院畜牧兽医研究所,福建福州350013

出　　处：《福建农业学报》2022年第11期1400-1406,共7页Fujian Journal of Agricultural Sciences

基　　金：福建省科技计划农业引导性(重点)项目(2019N0038)。

摘　　要：【目的】了解目前福建省莆田市鸭疫里默氏杆菌菌株对喹诺酮类相关耐药基因的携带情况,探讨其耐药机制。【方法】采用K-B法对2019–2021年分离自福建省莆田市的55株鸭疫里默氏杆菌菌株进行6种喹诺酮类药物的药敏试验,采用PCR方法对喹诺酮耐药决定区基因(QRDR)和质粒介导的喹诺酮耐药基因(PMQR)等6种耐药基因进行检测并分析。【结果】在55株鸭疫里默氏杆菌菌株中,50%以上菌株对吡哌酸、诺氟沙星、恩诺沙星、环丙沙星高度耐药,且为多重耐药;90%以上菌株对氧氟沙星高度敏感。耐药基因PCR检测结果显示,未检测到qnrA、qnrB、qnrS、oqxA等4种质粒介导的喹诺酮耐药基因。QRDR突变分析发现,gyrA基因推测氨基酸序列中有6个位点发生突变,突变类型有S83R和S83I、 S84P、 D87A和D87G、 A183V、 N202E、 Y211H;其中以S83I、N202E、Y211H的突变最多见,突变率分别为98.18%(54/55)、92.73%(51/55)、90.91%(50/55);39株耐恩诺沙星菌株中gyrA基因出现S83I(38/39)和S83R(1/39)突变,S83I突变也在3株恩诺沙星敏感菌株中发现。parC基因只在1株恩诺沙星中介菌株中出现F155S突变,这在鸭疫里默氏杆菌中未见报道。【结论】莆田市鸭疫里默氏杆菌的喹诺酮类药物耐药严重,未发现质粒介导的喹诺酮耐药基因,gyrA基因的S83I突变可能是介导鸭疫里默氏杆菌对喹诺酮类药物耐药的主要机制之一。【Objective】 Epidemiology and mechanism of quinolone-related drug resistance genes of Riemerella anatipestifer found in ducks in Putian, Fujian were investigated. 【Method】 The K-B method was applied to examine 55 strains of R.anatipestifer to determine their resistance to 6 types of quinolones. PCR was employed to detect and analyze the quinolone resistance determining region(QRDR) and plasmid mediated quinolone resistance(PMQR) genes. 【Result】 More than half of the 55 strains examined were multi-resistant and highly resistant to pipemidic acid, norfloxacin, enrofloxacin, and ciprofloxacin;and more than 90% of them highly sensitive to ofloxacin. PCR did not find the 4 PMQRs, i.e., qnrA, qnrB, qnrS,and oqxA, in the specimens. On the other hand, 6 QRDR mutations in the deduced amino acid sequences of gyrA, including S83R and S83I, S84P, D87A and D87G, A183V, N202E, and Y211H were detected. Among them, S83I, N202E, and Y211H had the highest mutation rates of 98.18%(54/55), 92.73%(51/55), and 90.91%(50/55), respectively. Of which, the S83I (38/39) and S83R(1/39) mutations were found in gyrA of 39 enrofloxacin-resistant strains, and the S83I mutations also in that of 3 enrofloxacin-sensitive strains. There was only one enrofloxacin-intermediated strain that had the F155S mutation in parC,which was not previously reported to exist in R. anatipestifer. 【Conclusion】 The quinolone-resistance of R. anatipestifer found in ducks in Putian was severe. No PMQR was detected in the samples, but the S83I mutation in gyrA of QRDR could be the major culprit that involved in the drug resistance of R. anatipestifer.

关 键 词：鸭疫里默氏杆菌 喹诺酮类 耐药基因 GYRA基因 PARC基因 

分 类 号：S852.61[农业科学—基础兽医学]