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目的建立一种基于Taqman探针的二重荧光定量PCR检测鼠疫菌方法,并进行应用评价。方法对收集的标本进行二重荧光定量PCR和鼠疫细菌学检测,然后进行统计分析。结果在209份样品中,二重荧光定量PCR检出鼠疫菌核酸阳性40份(19.14%),鼠疫菌分离培养检出阳性32份(15.31%),两种方法关联性有统计学意义(χ2=153.53,P<0.01),优势差异也有统计学意义(χ2=6.13,P<0.05)。结论本实验室所建立的二重荧光定量PCR检测鼠疫菌明显优于鼠疫菌分离培养,且操作简单、实验时间短,可作为鼠疫早期快速诊断方法。
Objective To establish a Taqman - based double fluorescence quantitative PCR method for the detection of Yersinia pestis and evaluate its application. Methods The collected samples were subjected to double fluorescence quantitative PCR and plague bacteriological examination, and then statistically analyzed. Results Of the 209 samples, 40 (19.14%) were positive for nucleic acid of Yersinia pestis and 32 (15.31%) were positive for Yersinia pestis isolated by double fluorescence quantitative PCR, the correlation between the two methods was statistically significant (χ2 = 153.53, P <0.01). There were also significant differences between the two groups (χ2 = 6.13, P <0.05). Conclusion The double fluorescence quantitative PCR assay established by our laboratory is obviously superior to Yersinia pestis isolates and cultured. It is easy to operate and has short experimental time, which can be used as an early rapid diagnostic method for plague.