【摘要】：銅綠假單胞菌(Pseudomonas aeruginosa,PA)是一種常見的人畜共患機(jī)會(huì)致病菌,給人畜健康帶來巨大威脅。本研究自臨床分離PA 51株,并對(duì)其耐藥表型、耐藥基因型、外毒素及動(dòng)物致病性進(jìn)行了檢測(cè)分析,以期為臨床銅綠假單胞菌病的研究提供基礎(chǔ)數(shù)據(jù)。自河北及周邊地區(qū)2011年-2014年臨床發(fā)病的雞、豬體內(nèi)分離并純化,通過形態(tài)學(xué)及PCR技術(shù)進(jìn)行鑒定后,獲得PA野毒51株;采用Kirby-Bauer紙片擴(kuò)散法(K-B法)進(jìn)行3類17種抗菌藥的藥物敏感性試驗(yàn),結(jié)果均表現(xiàn)為多重耐藥性;對(duì)氨基糖苷類藥物鏈霉素、阿米卡星、卡那霉素、新霉素、慶大霉素和大觀霉素的耐藥率依次為84.3%、84.3%、78.4%、78.4%、54.9%和25.2%;對(duì)喹諾酮類藥物氧氟沙星、恩諾沙星、鹽酸左氧、加替沙星、諾氟沙星和環(huán)丙沙星的耐藥率依次94.1%、82.4%、80.4%、78.4%、62.7%和25.2%;對(duì)β-內(nèi)酰胺類藥物頭孢拉定、青霉素、頭孢曲松鈉、頭孢噻肟、頭孢哌酮鈉/舒巴坦鈉的耐藥率依次98.0%、98.0%、70.6%、2.0%和0.0%。本文對(duì)51株P(guān)A分離株相應(yīng)的抗性基因進(jìn)行了PCR檢測(cè)。6種氨基糖苷類耐藥基因(AMEs)的檢測(cè)結(jié)果顯示,ant(3′′)-Ia、aac(6′)-Ib、aac(3)-IIa、aph(3′′)-Ib(str A)、aph(6′)-Id(str B)、Aph(3′)-IIa的陽性率由高到低依次為98.0%、51.0%、43.1%、41.2%、31.4%、17.6%;其中有1株為所測(cè)AMEs基因陰性,3株為6種基因全攜帶者;16Sr RNA甲基化酶(arm A、rmt A、rmt B)的檢測(cè)結(jié)果均為陰性;喹諾酮類耐藥質(zhì)粒的檢測(cè)陽性率由高到低依次為aac(6′)-Ib-cr(13.7%)、qnr S(9.8%)、qnr B(5.9%);對(duì)gyr A基因中的氟喹諾酮耐藥決定區(qū)(Quinolone-resisitant determining region,QRDR)進(jìn)行PCR檢測(cè)、測(cè)序、并進(jìn)行序列分析,結(jié)果僅3株QRDR基因在其編碼的第83位氨基酸發(fā)生突變,即83Thr→Ile;β-內(nèi)酰胺類耐藥基因的陽性率由高到低依次為OXA-3(7.8%)、OXA-1(2.0%)和OXA-5(0.0%)。PCR檢測(cè)結(jié)果顯示51株P(guān)A分離株細(xì)胞毒性外毒素A(Pseudomonas aeruginosa exo-toxin A,ETA)的陽性率為72.5%;動(dòng)物致病性回歸實(shí)驗(yàn)證明所分離的51株P(guān)A毒株均有較強(qiáng)的致病力。
[Abstract]:Pseudomonas aeruginosa (Pseudomonas aeruginosa,PA) is a common zoonosis opportunity pathogen, which brings great threat to human and animal health. In this study, 51 strains of PA were isolated from clinic, and their drug resistance phenotype, drug resistance genotype, exotoxin and animal pathogenicity were detected and analyzed in order to provide basic data for the study of clinical Pseudomonas aeruginosa disease. From Hebei and its surrounding areas from 2011 to 2014, pigs were isolated and purified in vivo. 51 strains of PA wild virus were obtained by morphology and PCR. The drug sensitivity test of three kinds of 17 kinds of antibiotics was carried out by Kirby-Bauer disk diffusion method (K 鈮，

本文編號(hào)：2479275