Enhanced Capacity Surge Laboratory, Reagent and Diagnostic Services Branch, Division of Scientific Resources, National Center for Emerging and Zoonotic Infectious Diseases (NCEZID) Centers for Disease Control & Prevention (CDC), Atlanta, Georgia.
Mycotic Diseases Branch, Division of Foodborne Waterborne and Environmental Diseases, National Center for Emerging and Zoonotic Infectious Diseases (NCEZID) Centers for Disease Control & Prevention (CDC), Atlanta, Georgia.
Mycoses. 2019 Jun;62(6):513-518. doi: 10.1111/myc.12907. Epub 2019 Apr 4.
Candida auris is an emerging multidrug-resistant yeast associated with invasive infection in healthcare settings. Recently, C auris cases in the United States have been detected in 11 states with the majority of cases in New York, New Jersey and Illinois. Rapid and accurate identification of C auris is critical for patient care and the implementation of public health measures to control the spread of infection. Our aim was to develop and validate a rapid DNA extraction method using the Roche MagNA Pure 96 instrument and a TaqMan real-time PCR assay for reliable, high-throughput identification of C auris. We evaluated 247 patient dermal swab samples previously analysed by culture/MALDI-TOF. The diagnostic sensitivity and specificity were 93.6% and 97.2%, respectively. The assay was highly reproducible with a detection limit of 1 C auris CFU/10 μL. A receiver operating characteristic curve analysis of the real-time PCR data showed an area of 0.982 under the curve, with a C cut-off value of ≤37.0. The turnaround time from DNA extraction to real-time PCR results was approximately 200 samples/day. In conclusion, we successfully validated a rapid and high-throughput method for accurate and reproducible identification of C auris with a significantly reduced turnaround time compared to culture/MALDI-TOF based methods.
耳念珠菌是一种新兴的多重耐药酵母菌，与医疗机构中的侵袭性感染有关。最近，美国 11 个州已经检测到了耳念珠菌病例，其中大多数病例在纽约、新泽西和伊利诺伊州。快速准确地识别耳念珠菌对于患者的治疗和实施公共卫生措施以控制感染的传播至关重要。我们的目的是开发和验证一种使用罗氏 MagNA Pure 96 仪器和 TaqMan 实时 PCR 检测的快速 DNA 提取方法，用于可靠、高通量地鉴定耳念珠菌。我们评估了 247 例先前通过培养/MALDI-TOF 分析的患者皮肤拭子样本。该检测方法的诊断灵敏度和特异性分别为 93.6%和 97.2%。该检测方法具有高度的可重复性，检测限为 1 个耳念珠菌 CFU/10μL。实时 PCR 数据的受试者工作特征曲线分析显示，曲线下面积为 0.982，C 截断值为≤37.0。从 DNA 提取到实时 PCR 结果的周转时间约为 200 个样本/天。总之，我们成功验证了一种快速、高通量的方法，可准确、可重复地鉴定耳念珠菌，与基于培养/MALDI-TOF 的方法相比，周转时间大大缩短。
