Q热贝纳柯克斯体TaqMan荧光定量PCR检测方法的建立

Establishment of Q Fever Coxiella burnetii TaqMan Fluorescent Quantitative PCR Detection Method

根据Q热贝纳柯克斯体(Coxiella burnetii)插入IS1111序列设计引物和探针,建立快速检测Q热的TaqMan实时荧光定量PCR方法。以梯度稀释含有目的扩增片段的重组质粒作为标准品,进行定量PCR反应。结果显示,该方法能够检测出10个拷贝数的阳性质粒;标准曲线相关系数为0.995,扩增效率为103%;结核分枝杆菌(M.tuberculosis)、衣原体(C.psittaci)、布鲁氏菌(Brucella.spp)及牛血液的核酸样本特异性检测结果均为阴性。本研究建立的TaqMan荧光定量PCR法灵敏度高、特异性好,对Q热的检测与鉴定中具有良好的应用前景。

Primers and probes were designed based on inserting sequence ISllll of Q fever Coxiella burnetii, TaqMan Real-time quantitative PCR assay was developed for indentification Q fever. The recombinant plasmid containing the target se- quence was constructed to detect the sensitivity and prepare the standard curve. The method could detect 10 of the plasmid cop- y number. Related coefficient was 0. 995 of the standard curve, the amplification efficiency was 103~. The results of specific detection of nucleic acid sample for M. tuberculosis, Brucella. spp, C. psittaci and bovine blood were negative. TaqMan fluores- cent quantitative PCR method was developed in this study and had high sensitivity and specificity. It had a good application prospects in the detection and identification of the Q fever.