分子检测对秋水仙碱耐药性的误诊:对秋水仙碱敏感的鲍曼不动杆菌分离物中 mcr-1-PCR 阳性的假象

IF 0.3 Q3 MEDICINE, GENERAL & INTERNAL Cukurova Medical Journal Pub Date : 2023-09-30 DOI:10.17826/cumj.1348548
Toğrul NAĞIYEV, Tülay KANDEMİR, Fatih KÖKSAL
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 Materials and Methods: The mcr-1 gene was examined using PCR in 103 carbapenem-resistant isolates, including 75 Acinetobacter baumannii, 19 Pseudomonas aeruginosa, and 9 Klebsiella pneumoniae. DNA sequencing was performed to confirm the mcr-1 positivity. Other antimicrobial resistance genes were investigated in isolates that were found to be mcr-1-positive by PCR and colistin-resistant isolates. 
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引用次数: 0

摘要

目的:本研究的目的是调查mcr-1基因的存在,这是负责粘菌素耐药,在碳青霉烯耐革兰氏阴性菌引起难以治疗的感染在土耳其的一家研究医院。& # x0D;材料与方法:采用PCR方法检测103株碳青霉烯耐药菌株的mcr-1基因,包括75株鲍曼不动杆菌、19株铜绿假单胞菌和9株肺炎克雷伯菌。DNA测序证实mcr-1阳性。对PCR检测mcr-1阳性的分离株和耐粘菌素的分离株进行了其他抗菌素耐药基因的研究。& # x0D;结果103株碳青霉烯类耐药菌株中有3.9%、75株鲍曼不动杆菌中有5.3%对粘菌素敏感,4株鲍曼不动杆菌耐药菌株(鲍曼不动杆菌1株、肺炎克雷伯菌3株)中均未检出mcr-1。DNA测序分析结果表明,扩增产物均不是目标片段,但与鲍曼不饱和杆菌的染色体DNA片段匹配度超过70%。因此,这些结果被认为是假阳性。虽然这些假阳性分离株对粘菌素敏感,但它们具有广泛耐药(XDR)。其中2只携带blaOXA23-like和blaTEM基因,另1只携带blaOXA23-like、blaTEM和blaOXA48-like基因,第4只携带blaOXA23-like和blaCTXM基因。& # x0D;结论:虽然大多数研究报道PCR检测mcr-1基因的引物特异性为100%,但我们认为PCR检测在快速常规诊断中尚不足以单独使用或与抗生素药敏试验联合使用。至少在一段时间内,使用DNA序列分析确认pcr阳性样本是合适的。
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Kolistin direncinin moleküler tespitinde yanlış tanı: kolistine duyarlı Acinetobacter baumannii izolatlarında yanlış mcr-1-PCR pozitifliği
Purpose: The aim of this study was to investigate the presence of the mcr-1 gene, which is responsible for colistin resistance, in carbapenem-resistant Gram-negative bacteria that cause difficult-to-treat infections in a research hospital in Turkey. Materials and Methods: The mcr-1 gene was examined using PCR in 103 carbapenem-resistant isolates, including 75 Acinetobacter baumannii, 19 Pseudomonas aeruginosa, and 9 Klebsiella pneumoniae. DNA sequencing was performed to confirm the mcr-1 positivity. Other antimicrobial resistance genes were investigated in isolates that were found to be mcr-1-positive by PCR and colistin-resistant isolates. Results: Four (3.9% of the 103 carbapenem-resistant isolates and 5.3% of the 75 A. baumannii isolates) A. baumannii isolates, all susceptible to colistin, were found to be mcr-1-positive by PCR, whereas mcr-1 was not detected in four colistin-resistant isolates, one in A. baumannii and three in K. pneumoniae. DNA sequencing analysis determined that none of the amplification products was the targeted fragment, but they matched more than 70% with the chromosomal DNA fragments of A. baumannii strains. Therefore, these results were considered false-positive. Although these false-positive isolates were susceptible to colistin, they were extensively drug-resistant (XDR). Two of them were found to carry blaOXA23-like and blaTEM genes, another blaOXA23-like, blaTEM and blaOXA48-like genes, and the fourth one to have blaOXA23-like and blaCTXM genes. Conclusion: Although the specificity of the primers used to detect the mcr-1 gene by PCR was reported as 100% in most studies, we concluded that PCR tests are insufficient yet to use alone or with antibiotic susceptibility tests in rapid routine diagnosis. Confirming at least PCR-positive samples using DNA sequence analysis would be appropriate for a certain period.
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Cukurova Medical Journal
Cukurova Medical Journal MEDICINE, GENERAL & INTERNAL-
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发文量
159
审稿时长
12 weeks
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