检测gyrA和parC突变以及质粒介导喹诺酮耐药基因的流行情况于…… （原文未完整给出地点等信息）
Detection of gyrA and parC Mutations and Prevalence of Plasmid-Mediated Quinolone Resistance Genes in .
作者信息
Kareem Sawsan Mohammed, Al-Kadmy Israa M S, Kazaal Saba S, Mohammed Ali Alaa N, Aziz Sarah Naji, Makharita Rabab R, Algammal Abdelazeem M, Al-Rejaie Salim, Behl Tapan, Batiha Gaber El-Saber, El-Mokhtar Mohamed A, Hetta Helal F
机构信息
Branch of Biotechnology, Department of Biology, College of Science, University of Mustansiriyah, Baghdad, Iraq.
Faculty of Science & Engineering, School of Engineering, University of Plymouth, Plymouth, PL4 8AA, UK.
出版信息
Infect Drug Resist. 2021 Feb 12;14:555-563. doi: 10.2147/IDR.S275852. eCollection 2021.
BACKGROUND AND AIM
Recently, the extensive use of quinolones led to increased resistance to these antimicrobial agents, with different rates according to the organism and the geographical region. The aim of this study was to detect the resistance rate of Iraqi isolates toward quinolone antimicrobial agents, to determine genetic mutations in and , to screen for efflux-pump activity, and to screen the presence of plasmid-mediated quinolone resistance (PMQR) genes.
METHODS
Forty-three isolates were confirmed phenotypically and genotypically by Vitek 2 system and species specific primers by PCR using the targeting gene followed by sequencing. Antibiotic susceptibility test was carried out using disc diffusion method. Quinolone resistant isolates were subjected to ciprofloxacin MIC testing, and cartwheel method to screen for efflux pump activity. The presence of the plasmid mediated quinolone resistance genes , and was tested by PCR. Sequencing of A and C was performed.
RESULTS
We observed a high rate of resistance to ceftriaxone, gentamicin ciprofloxacin, and levofloxacin. Low rate of resistance was detected against amikacin and azithromycin. Ciprofloxacin MIC results revealed that 96.1% of the isolates had MICs >256 µg/mL, 83.4% had MICs >512 µg/mL while 34.6% had MIC >1024 µg/mL. Testing of isolates against ciprofloxacin mixed with EtBr at various concentrations resulted in decreased resistant. Sequencing results showed that Ser83Leu was the most common mutation in A that was observed in all quinolone resistant isolates, followed by Asp87Asn. Ser80Ile mutation in C was observed in 77.7% of the tested isolates. The prevalence of PMQR genes was 92.5% , 51.8% B, 40.7% A, and 37% S.
CONCLUSION
Quinolone resistance is common in isolates in Baghdad. The frequent mutation in A and C, and the presence of PMQR genes is alarming.
背景与目的
近期，喹诺酮类药物的广泛使用导致对这些抗菌药物的耐药性增加，不同生物体和地理区域的耐药率各异。本研究的目的是检测伊拉克分离株对喹诺酮类抗菌药物的耐药率，确定gyrA和parC中的基因突变，筛查外排泵活性，并筛查质粒介导的喹诺酮耐药（PMQR）基因的存在情况。
方法
通过Vitek 2系统以及使用靶向gyrA基因的物种特异性引物进行PCR并测序，对43株分离株进行表型和基因型确认。采用纸片扩散法进行药敏试验。对喹诺酮耐药分离株进行环丙沙星最低抑菌浓度（MIC）检测，并采用车轮法筛查外排泵活性。通过PCR检测质粒介导的喹诺酮耐药基因qnrA、qnrB、qnrS和aac(6’)-Ib-cr的存在情况。对gyrA和parC进行测序。
结果
我们观察到分离株对头孢曲松、庆大霉素、环丙沙星和左氧氟沙星的耐药率较高。对阿米卡星和阿奇霉素的耐药率较低。环丙沙星MIC结果显示，96.1%的分离株MIC>256μg/mL，83.4%的分离株MIC>512μg/mL，而34.6%的分离株MIC>1024μg/mL。对分离株进行不同浓度环丙沙星与溴化乙锭混合检测，耐药性降低。测序结果显示，Ser83Leu是gyrA中在所有喹诺酮耐药分离株中最常见的突变，其次是Asp87Asn。在77.7%的受试分离株中观察到parC中的Ser80Ile突变。PMQR基因的流行率为qnrB 92.5%、qnrA 51.8%、qnrS 40.7%和aac(6’)-Ib-cr 37%。
结论
喹诺酮耐药在巴格达的分离株中很常见。gyrA和parC中的频繁突变以及PMQR基因的存在令人担忧。
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