实时荧光定量PCR快速检测脑膜炎奈瑟菌的临床价值

Real-time fluorescence quantitation polymerase chain reaction for Neisseria meningitidis determination

目的探讨实时荧光定量聚合酶链反应(PCR)快速检测流行性脑脊髓膜炎(简称流脑)病原菌脑膜炎奈瑟菌(Nm)的临床价值。方法收集70例疑似流脑病例的双份脑脊液标本,1份采用细菌培养法进行检测,另1份采用实时荧光定量PCR进行检测,比较2种方法的检出阳性率以及敏感性和特异性。结果采用细菌培养法检出Nm阳性49例,阳性率为70.00%;实时荧光定量PCR检出Nm阳性67例,阳性率为95.71%,二者阳性率比较差异有统计学意义(P<0.05)。实时荧光定量PCR检测结果与细菌培养法阳性符合率为70.00%,其中有18例细菌培养法为阴性而实时荧光定量PCR为阳性。实时荧光定量PCR检测Nm的特异性为100%,无假阴性结果,阳性预测值为73.13%,阴性预测值为0.00%,检测下限为103拷贝/m L,线性范围为2×103~2×107拷贝/m L。结论实时荧光定量PCR检测Nm敏感性高、特异性强,且快速简便,为流脑流行病学调查提供了一种快速、简便的病原学诊断手段。

Objective To study the role of real-time fluorescence quantitation polymerase chain reaction(PCR)for Neisseria meningitidis(Nm)determination.Methods A total of 70 cerebrospinal fluid specimens were collected from patients with suspected meningitis.One part was performed bacterial culturing,and another part was determined by real-time fluorescence quantitation PCR.The positive rates,sensitivities and specificities of the 2 methods were compared.Results The positive rate of Nm by bacterial culturing was 70.00%(49 cases).Realtime fluorescence quantitation PCR showed a positive rate of 95.71%(67 cases).There was statistical significance for the positive rates between the 2 methods(P<0.05).The results of real-time fluorescence quantitation PCR and bacterial culturing showed a consistency rate of 70.00%.There were 18 cases being negative by bacterial culturing but being positive by real-time fluorescence quantitation PCR.The specificity of real-time fluorescence quantitation PCR was 100%,and no false negative results were obtained.The positive and negative predictive values were 73.13%and 0.00%,respectively.The lower limit of determination was 103 copies/mL.The linear range was 2×103-2×107 copies/L.Conclusions Real-time fluorescence quantitation PCR for Nm determination has high sensitivity and specificity and is rapid and convenient,which can be used for the etiological diagnosis of meningitis epidemiology.