He Yuanhui, Ruan Genjie, Hao Hui, Xue Feng, Zhu Sainan, Xiao Bingbing, Zheng Bo
Institute of Clinical Pharmacology, Peking University First Hospital, Beijing, China.
Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing, China.
Front Med (Lausanne). 2020 Sep 8;7:403. doi: 10.3389/fmed.2020.00403. eCollection 2020.
To evaluate the sensitivity and accuracy of the quadruple real-time PCR method for the detection of enterococci carrying vancomycin-resistant genes , and in rectal swabs. Choosing PCR-sequenced DNA extracted directly from rectal swabs as the gold standard, the results of the quadruple real-time PCR method and the traditional method (screening culture combined PCR-sequencing method whose DNA extracted from single colony) were compared with the gold standard. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of the quadruple real-time PCR method and the traditional method were obtained. The time required for the three methods was calculated. The results between gold standard and the quadruple real-time PCR method were similar. Compared to the traditional method, the quadruple real-time PCR method had a much higher sensitivity, specificity, PPV, NPV, and consistency. Our study found that the quadruple real-time PCR method is beneficial for detection of enterococci carrying with vancomycin heteroresistance. The traditional method had high specificity and NPV, but its sensitivity and PPV were not ideal. The time needed for gold standard is a minimum of 28 h; the quadruple real-time PCR method takes 2-3 h while the traditional method consumes a minimum of 72 h. The quadruple real-time PCR method can provide a rapid and reliable result for the diagnosis of patients with colonized vancomycin-resistant enterococci. This new method is beneficial for the active screening, timely clinical treatment measures, epidemiological research, and hospital monitoring of the enterococci carrying vancomycin-resistant gene, especially for the enterococci with vancomycin heteroresistance carrying .
为评估四重实时荧光定量PCR法检测携带耐万古霉素基因的肠球菌及直肠拭子中的肠球菌的敏感性和准确性。以直接从直肠拭子中提取的经PCR测序的DNA作为金标准，将四重实时荧光定量PCR法和传统方法（从单菌落提取DNA的筛选培养联合PCR测序法）的结果与金标准进行比较。得出四重实时荧光定量PCR法和传统方法的敏感性、特异性、阳性预测值（PPV）、阴性预测值（NPV）及准确性。计算三种方法所需时间。金标准与四重实时荧光定量PCR法的结果相似。与传统方法相比，四重实时荧光定量PCR法具有更高的敏感性、特异性、PPV、NPV及一致性。我们的研究发现，四重实时荧光定量PCR法有助于检测携带万古霉素异质性耐药的肠球菌。传统方法具有较高的特异性和NPV，但其敏感性和PPV并不理想。金标准所需时间最少为28小时；四重实时荧光定量PCR法需2 - 3小时，而传统方法最少需72小时。四重实时荧光定量PCR法可为耐万古霉素肠球菌定植患者的诊断提供快速可靠的结果。这种新方法有利于对携带耐万古霉素基因的肠球菌，尤其是携带万古霉素异质性耐药的肠球菌进行主动筛查、及时的临床治疗措施、流行病学研究及医院监测。
