Q热贝纳柯克斯体TaqMan荧光定量PCR检测方法的建立

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。

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

Primers and probes were designed based on inserting sequence IS1111 of Q fever Coxiella burnetii, TaqMan Real-time quantitative PCR assay was developed for indentification Q fever. The recombinant plasmid containing the target sequence was constructed to detect the sensitivity and prepare the standard curve. The method could detect 10 of the plasmid copy 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 fluorescent 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.