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应用 PCR-限制性片段长度多态性分析方法评估禽源和人源空肠弯曲菌和结肠弯曲菌中的氟喹诺酮耐药性。

Fluoroquinolone resistance in Campylobacter jejuni and Campylobacter coli from poultry and human samples assessed by PCR-restriction fragment length polymorphism assay.

机构信息

Center for Diagnosis and Research on Avian Pathology (CDPA), Faculty of Veterinary Medicine, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Brazil.

Faculty of Veterinary Medicine, University of Passo Fundo (UPF), Passo Fundo, Brazil.

出版信息

PLoS One. 2018 Jul 6;13(7):e0199974. doi: 10.1371/journal.pone.0199974. eCollection 2018.

Abstract

The objective of this study was to determine fluoroquinolone resistance in Campylobacter spp from poultry and human isolates. Forty-one Campylobacter jejuni isolates (30 of poultry origin and 11 of human origin) and 11 Campylobacter coli isolates (10 of human origin and 1 of poultry origin) were examined for ciprofloxacin, norfloxacin, and nalidixic acid resistance using the minimal inhibitory concentration (MIC) method. Thereafter, the isolates were analyzed by PCR-Restriction Fragment Length Polymorphism (RFLP) assay for detection of Thr-86 mutation. Finally, DNA sequencing was performed for confirmation of gyrA gene mutation. A complete correlation was observed between MICs, PCR-RFLP assay, and sequencing. The results revealed high quinolone resistance rates for C. jejuni (100%) and C. coli (100%) isolates obtained from poultry and moderate resistance for C. jejuni (9.1%) and C. coli (40%) samples of human origin. A mutation in codon 86 of the gyrA gene with a Thr-to-Ile substitution is reported to be the main cause of high resistance to quinolones. This mutation can be analyzed by PCR-RFLP assay, which has been proven to be a simple and fast method for the detection of fluoroquinolone resistance in Campylobacter spp.

摘要

本研究旨在确定禽源和人源弯曲杆菌分离株中的氟喹诺酮耐药性。使用最小抑菌浓度(MIC)法检测了 41 株空肠弯曲杆菌分离株(30 株来自禽类,11 株来自人类)和 11 株大肠弯曲杆菌分离株(10 株来自人类,1 株来自禽类)对环丙沙星、诺氟沙星和萘啶酸的耐药性。然后,通过 PCR-限制性片段长度多态性(RFLP)分析检测 Thr-86 突变来分析分离株。最后,进行 DNA 测序以确认 gyrA 基因突变。MIC、PCR-RFLP 分析和测序之间完全相关。结果表明,来自禽源的空肠弯曲杆菌(100%)和大肠弯曲杆菌(100%)分离株的喹诺酮耐药率较高,而来自人源的空肠弯曲杆菌(9.1%)和大肠弯曲杆菌(40%)的分离株的耐药率适中。报道称,gyrA 基因 86 密码子处的 Thr-to-Ile 取代突变是导致喹诺酮类药物高度耐药的主要原因。该突变可通过 PCR-RFLP 分析进行分析,该分析已被证明是检测弯曲杆菌属中氟喹诺酮类耐药性的一种简单快速的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3267/6034818/e237770f3687/pone.0199974.g001.jpg

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