【摘要】 目的探讨实时荧光定量PCR(real-time fluorescent quantitative polymerase chain reaction,RT-FQ-PCR)法检测新生隐球菌感染的临床应用价值。方法选择2014年2月-2014年7月就诊的47例疑似新生隐球菌感染患者的血液标本,采用常规培养基培养方法以及RT-FQ-PCR法进行检测,计算检测新生隐球菌的阳性率,并进行统计学分析。采用RT-FQ-PCR法对新生隐球菌、光滑念珠菌、克柔念珠菌、烟曲霉、黑曲霉、大肠杆菌、肺炎球菌、金黄色葡萄球菌和链球菌这些常见真菌和细菌的DNA进行检测,评价RT-FQ-PCR法的特异性。采用RT-FQ-PCR和常规培养基培养法对稀释至5个/m L的新生隐球菌菌液进行检测,评价RT-FQ-PCR法的灵敏度。结果采用RT-FQ-PCR法,以新生隐球菌特异性引物为探针对新生隐球菌和其他常见真菌及细菌进行检测,只有新生隐球菌出现荧光信号。RTFQ-PCR在5个/m L浓度的新生隐球菌菌液中检测出新生隐球菌,而常规培养基培养法未检测出。在47例疑似新生隐球菌感染患者中,常规培养基培养方法检测出11例,阳性率为23.40%,而RT-FQPCR法检测出14例,阳性率为29.79%,二者经比较差异无统计学意义(P>0.05)。结论 RT-FQ-PCR法与常规培养基培养法对新生隐球菌的检测具有很好的一致性。且RT-FQ-PCR法能够检测出较低浓度的新生隐球菌,较常规培养基培养法快速、灵敏度高、特异性强,可缩短报告时间,为临床早期诊断及治疗新生隐球菌感染提供可靠依据。更多还原

【Abstract】 Objective To investigate the clinical application value of real-time fluorescent quantitative polymerase chain reaction(RT-FQ-PCR) method for Cryptococcus neoformans detection in blood. Methods The blood samples of 47 patients with probable Cryptococcus neoformans infection were selected from February 2014 to July 2014 in hospital. The blood samples were measured by routine culture method and detected DNA fragments by RT-FQ-PCR method. And the detection rates were evaluated by statistical method.Cryptococcus neoformans, Candida glabrata, Candida krusei, Aspergillus fumigatus, Aspergillus niger, Escherichia coli, Streptococcus pneumoniae, Staphylococcus aureus and Streptococcus were detected by RT-FQ-PCR method to evaluate the specificity. The concentration of Cryptococcus neoformans solution was detected by RT-FQ-PCR method and routine culture method to evaluate the sensitivity. Results In RT-FQ-PCR method detection, only Cryptococcus neoformans had fluorescence signal. In 5 / m L concentration of Cryptococcus neoformans solution, Cryptococcus neoformans was only detected by RT-FQ-PCR method. In 47 cases probable Cryptococcus neoformans infection patients, positive rate of routine culture method was 23.40%(11cases), and positive rate of RT-FQ-PCR method was 29.79%(14 cases). The difference had no statistical significance(P > 0.05). Conclusion RT-FQ-PCR method and routine culture method have a higher concordance for Cryptococcus neoformans detection. RT-FQ-PCR method can detect low concentration of Cryptococcus neoformans. Compared with routine culture method, RT-FQ-PCR has shorter time, higher sensitivity and specificity, which can reduce report time and provide reliable basis for clinical early diagnosis and treatment of Cryptococcus neoformans infection.更多还原