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首页> 外文期刊>Journal of Clinical Microbiology >Development and Validation of a Real-Time PCR Assay for Rapid Detection of Candida auris from Surveillance Samples
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Development and Validation of a Real-Time PCR Assay for Rapid Detection of Candida auris from Surveillance Samples

机译:用于从监测样品中快速检测金眼假丝酵母的实时PCR检测方法的开发和验证

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ABSTRACT Candida auris is an emerging multidrug-resistant yeast causing invasive health care-associated infection with high mortality worldwide. Rapid identification of C. auris is of primary importance for the implementation of public health measures to control the spread of infection. To achieve these goals, we developed and validated a TaqMan-based real-time PCR assay targeting the internal transcribed spacer 2 ( ITS 2) region of the ribosomal gene. The assay was highly specific, reproducible, and sensitive, with the detection limit of 1 C. auris CFU/PCR. The performance of the C. auris real-time PCR assay was evaluated by using 623 surveillance samples, including 365 patient swabs and 258 environmental sponges. Real-time PCR yielded positive results from 49 swab and 58 sponge samples, with 89% and 100% clinical sensitivity with regard to their respective culture-positive results. The real-time PCR also detected C. auris DNA from 1% and 12% of swab and sponge samples with culture-negative results, indicating the presence of dead or culture-impaired C. auris . The real-time PCR yielded results within 4 h of sample processing, compared to 4 to 14 days for culture, reducing turnaround time significantly. The new real-time PCR assay allows for accurate and rapid screening of C. auris and can increase effective control and prevention of this emerging multidrug-resistant fungal pathogen in health care facilities.
机译:摘要假丝酵母是一种新兴的多药耐药性酵母,可引起侵入性医疗相关感染,在全球范围内具有很高的死亡率。快速识别金黄色葡萄球菌对于实施控制感染传播的公共卫生措施至关重要。为了实现这些目标,我们开发并验证了基于TaqMan的实时PCR分析方法,该方法针对核糖体基因的内部转录间隔区2(ITS 2)区。该测定法具有高度特异性,可重复性和灵敏性,检测限为1 C. auris CFU / PCR。通过使用623个监测样本(包括365个患者拭子和258个环境海绵)来评估金黄色葡萄球菌实时PCR分析的性能。实时PCR从49个拭子和58个海绵样品中获得了阳性结果,就其各自的培养阳性结果而言,临床敏感性为89%和100%。实时PCR还从1%和12%的拭子和海绵样品中检测到金黄色葡萄球菌DNA,培养阴性,表明存在死的或受损的金黄色葡萄球菌。与培养4到14天相比,实时PCR在样品处理后4小时内产生了结果,从而大大缩短了周转时间。新的实时PCR测定法可准确快速地筛查金黄色葡萄球菌,并可以在医疗保健机构中有效控制和预防这种新兴的多药耐药真菌病原体。

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