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首页> 外文期刊>Journal of Clinical Microbiology >Enhancing Gonococcal Antimicrobial Resistance Surveillance: a Real-Time PCR Assay for Detection of Penicillinase-Producing Neisseria gonorrhoeae by Use of Noncultured Clinical Samples
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Enhancing Gonococcal Antimicrobial Resistance Surveillance: a Real-Time PCR Assay for Detection of Penicillinase-Producing Neisseria gonorrhoeae by Use of Noncultured Clinical Samples

机译：加强淋球菌抗菌素耐药性监测：实时PCR检测非培养临床样品检测生产青霉素酶淋病奈瑟氏球菌的方法

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With increasing concerns regarding diminishing treatment options for gonorrhea, maintaining the efficacy of currently used treatments and ensuring optimal Neisseria gonorrhoeae antimicrobial resistance surveillance are of the utmost importance. Penicillin is still used to treat gonorrhea in some parts of the world. In this study, we developed and validated a real-time PCR assay for the detection of penicillinase-producing N. gonorrhoeae (PPNG) in noncultured clinical samples with the aim of enhancing penicillin resistance surveillance. The assay (PPNG-PCR2) was designed to be an indirect marker of penicillinase activity, by targeting a region of sequence predicted to be conserved across all N. gonorrhoeae plasmid types harboring the beta-lactamase gene while not specifically targeting the actual beta-lactamase-encoding sequence. The assay was evaluated by using a total of 118 N. gonorrhoeae clinical isolates and 1,194 clinical specimens, including 239 N. gonorrhoeae-positive clinical samples from which N. gonorrhoeae cells were isolated and for which phenotypic penicillinase results are available. Overall, the PPNG-PCR2 assay provided 100% sensitivity and 98.7% specificity compared to bacterial culture results for the detection of PPNG in clinical specimens. PPNG-PCR2 false-positive results, presumably due to cross-reactions with unrelated bacterial species, were observed for up to 1.3% of clinical samples but could be distinguished on the basis of high cycle threshold values. In tandem with phenotypic surveillance, the PPNG-PCR2 assay has the potential to provide enhanced epidemiological surveillance of N. gonorrhoeae penicillin resistance and is of particular relevance to regions where penicillin is still used to treat gonorrhea.
机译：对于减少淋病的治疗选择的关注日益增加，保持当前使用的治疗方法的有效性以及确保最佳的淋病奈瑟氏球菌的抗菌素耐药性监测至关重要。在世界某些地区，青霉素仍用于治疗淋病。在这项研究中，我们开发并验证了一种实时PCR检测方法，用于检测非培养的临床样本中产生青霉素酶的淋病奈瑟氏球菌（PPNG），目的是增强对青霉素的耐药性监测。通过靶向预计在所有携带β-内酰胺酶基因的淋病奈瑟氏球菌质粒类型中均保守的序列区域，而不是专门针对实际的β-内酰胺酶，将测定法（PPNG-PCR2）设计为青霉素酶活性的间接标记-编码序列。通过使用总共118株淋病奈瑟氏球菌临床分离株和1194份临床标本来评估该分析方法，其中包括239株淋病奈瑟氏球菌阳性临床样本，从中分离出淋病奈瑟球菌细胞并获得表型青霉素酶结果。总体而言，与细菌培养结果相比，PPNG-PCR2检测可提供100％的敏感性和98.7％的特异性，可用于检测临床标本中的PPNG。 PPNG-PCR2假阳性结果可能是由于与无关细菌物种的交叉反应所致，最多可观察到1.3％的临床样品，但可以根据高循环阈值进行区分。与表型监测相结合，PPNG-PCR2检测具有增强淋病奈瑟氏球菌对青霉素耐药性的流行病学监测的潜力，并且与仍使用青霉素治疗淋病的地区特别相关。

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