荧光定量PCR方法检测人群血样中的布鲁杆菌DNA

Detection of Brucella spp. DNA in patients serum samples by real-time PCR

目的评估荧光定量PCR方法检测人群血样中布鲁杆菌DNA的快速性和特异性。方法以bcsp31基因作为荧光定量PCR的检测目标，利用15种已知布鲁杆菌菌株和41种非布鲁杆菌菌株DNA评价荧光定量PCR方法检测布鲁杆菌的种属特异性。以2012年哈尔滨市疾病预防控制中心布鲁杆菌病监测人群结果中的17份布鲁杆菌阳性血液为样本，另选取30名健康者和30名非布鲁杆菌病（布病）患者血样作为对照．评价荧光定量PCR方法应用于检测人群血样布鲁杆菌的相应灵敏度和特异性。结果荧光定量．PCR结果显示15个布鲁杆菌中bcsp31基因检测均为阳性．41个非布鲁杆菌种均未检出bcsp31基因。在17名布病患者中，11份血液样本bcsp31基因扩增信号阳性，灵敏度为64．7％（11／17）；60名对照的血清均显示阴性结果，相应灵敏度为100．0％。结论与传统方法相比，荧光定量PCR方法具有快速、特异等优点，适用于布病确诊。

Objective In this article we evaluated the sensitivities and specificities of real-time PCR assay for diagnosis of human brucellosis. Methods The species selectivity and specificity of real-time PCR were evaluated by direct amplification of a 169 bp portion of bcsp31 gene from 15 Brucella strains and 41 non-Brucella strains. Aceording to the monitoring results of 2012 Harbin brucellosis, 17 brucellosis patients and 30 health people were selected to collect their serum samples for assessing the sensitivity of real-time PCR, and additional 30 non- brucellosis patients serum samples were as eontrols. Results The speeies seleetivity and specificity of our real- time PCR method were evaluated by using genomic DNA from 15 Brucella strains and 41 non-Brucella strains. There were 11 sera with positive amplification signals among the 17 culture-proven brucellosis patients, the sensitivity was 64.7% （11/17）. Whereas, the results of sera from the 60 control patients were all negative, corresponding to a specificity of 100.0%. Conclusion The results indicate that real-time PCR is well suitable for confirmation of brucellosis cases.