铜绿假单胞菌是一种非发酵革兰阴性杆菌。许多毒力因子在铜绿假单胞菌的发病机制中起作用。这项研究的目的是通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOFMS)方法早期检测ST111,ST175,ST235,ST253,ST395,这些克隆被命名为高风险克隆,由于铜绿假单胞菌分离物中的多药耐药性而具有增加的流行潜力(MALDI-TOFMS)方法,并评估高风险分离物与存在之间的关系。对于毒力基因的检测,用于toxA的单重聚合酶链反应(PCR)和用于algD的多重PCR,plcN,LasB,plcH在铜绿假单胞菌分离株中进行。在碳青霉烯酶基因的检测中,用于BlaKPC的两个单独的多路复用PCR,blaNDM,BlaVIM,blaOXA-48和blaIMP,blaSPM,blaSIM,BlaGIM,blages.通过使用VITEK®-MS(bioMérieux,法国)系统。铜绿假单胞菌分离株主要来自重症监护病房(45%)和呼吸道样本(46%)。发现分离株最敏感的抗生素是阿米卡星,而哌拉西林的耐药性最高。在PCR结果中,toxa,LasB,plcH,plcN和algD检测为89%,99%,98%,100%,100%,分别。当使用MALDI-TOFMS评估属于高风险克隆的特征峰的存在时,检测到ST253(7%)和ST175(2%)。在我们的研究中未检测到对ST235和ST395克隆特异的峰。在两个分离物中检测到blaVIM,在两个分离物中检测到blaGES-5碳青霉烯酶。在高风险克隆和其他菌株中均检测到高比率的毒力因子,并且在高风险克隆和毒力因子之间没有发现显着关系。及早发现高危克隆,确定抗菌素耐药机制将有助于制定战略治疗方案并防止其在全球范围内传播。
Pseudomonas aeruginosa is a non-fermentative gram-negative bacillus. Many virulence factors play a role in the pathogenesis of P.aeruginosa. The aim of this study was to early detection of ST111, ST175, ST235, ST253, ST395 which are named high-risk clones with increased epidemic potential due to multidrug resistance in P.aeruginosa isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method and to evaluate the relationship between high-risk clones and the presence of P.aeruginosa virulence factors and carbapenemase production genes.P.aeruginosa isolates (n= 100) found to be resistant to at least
imipenem or meropenem antibiotics isolated from the various clinical samples in the medical microbiology laboratory between 01.01.2021 and 07.06.2022 were included in the study. For the detection of virulence genes uniplex polymerase chain reaction (PCR) for toxA and multiplex PCR for algD, plcN, lasB, plcH were performed in P.aeruginosa isolates. In the detection of carbapenemase genes, two separate multiplex PCRs used for blaKPC , blaNDM , blaVIM , blaOXA-48 and for blaIMP , blaSPM , blaSIM , blaGIM , blaGES . Investigation of the peaks specific to high-risk clones was performed by using VITEK®-MS (bioMérieux, France) system. P.aeruginosa isolates were mostly isolated from intensive care units (45%) and respiratory tract samples (46%). The antibiotic to which the isolates were found to be most susceptible was amikacin, while highest resistance was detected for piperacillin. In PCR results, toxA, lasB, plcH, plcN and algD were detected as 89%, 99%, 98%, 100%, 100%, respectively. When the presence of characteristic peaks belonging to high-risk clones was evaluated with MALDI-TOF MS, ST253 (7%) and ST175 (2%) were detected. The peaks specific to ST235 and ST395 clones were not detected in our study. blaVIM was detected in two isolates and blaGES-5 carbapenemase was detected in two isolates. Virulence factors were detected at high rates in both high-risk clones and other strains and no significant relationship was found between high-risk clones and virulence factors. Early detection of high-risk clones, identification of antimicrobial resistance mechanisms will help to develop strategic treatment options and prevent their worldwide spread.