Development and Validation of a Real-Time PCR Assay for Rapid Detection of Candida auris from Surveillance Samples.
Development and Validation of a Real-Time PCR Assay for Rapid Detection of Candida auris from Surveillance Samples.
复制标题
从监视样品中快速检测到念珠菌的实时PCR分析的开发和验证。
DOI：
10.1128/jcm.01223-17
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发表时间：
2018-03
影响因子：
9.4
通讯作者：
Chaturvedi S
中科院分区：
文献类型：
--
作者：
Leach L;Zhu Y;Chaturvedi S
耳念珠菌是一种新兴的多重耐药酵母菌，可引起侵入性医疗保健相关感染，并在全球范围内造成高死亡率。快速识别耳念珠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，我们开发并验证了一种基于 TaqMan 的实时 PCR 检测，针对核糖体基因的内部转录间隔区 2 (ITS2) 区域。该测定具有高度特异性、可重复性和敏感性，检测限为 1 个耳念珠菌 CFU/PCR。使用 623 个监测样本（包括 365 个患者拭子和 258 个环境海绵）评估了耳念珠菌实时 PCR 检测的性能。实时 PCR 从 49 个拭子和 58 个海绵样本中得出阳性结果，其各自的培养阳性结果的临床敏感性分别为 89% 和 100%。实时 PCR 还从 1% 和 12% 的拭子和海绵样本中检测到耳念珠菌 DNA，且培养结果呈阴性，表明存在死亡或培养受损的耳念珠菌。实时 PCR 在样品处理后 4 小时内即可得出结果，而培养则需要 4 至 14 天，从而显着缩短了周转时间。新的实时 PCR 检测可以准确、快速地筛查耳念珠菌，并可以在医疗保健机构中加强对这种新兴多重耐药真菌病原体的有效控制和预防。
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 (ITS2) 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.