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目的:建立登革病毒的荧光定量PCR检测技术。方法:针对1~4型登革病毒3′端非编码区的一段高度保守序列设计引物和探针,建立登革病毒实时荧光PCR检测方法及定量的检测方法。并对10例ELISA法检测为登革病毒阳性的病例,取其发热1~2天的临床血清标本进行荧光定量PCR检测。结果:实时荧光PCR检测1~4型登革病毒均为阳性,该探针和引物是登革病毒检测的通用探针和引物。实时荧光PCR检测10份临床血清,6份为阳性,阳性率为60%。结论:实时PCR方法是一个快速、特异性强、敏感性高的检测登革病毒的方法,适用于登革热的临床早期诊断,具有很好的社会效益和经济效益。
Objective: To establish a dengue virus fluorescence quantitative PCR detection technology. Methods: Primers and probes were designed according to a highly conserved sequence of the noncoding region of dengue virus type 1 ~ 4, and a real - time fluorescent PCR detection method and a quantitative detection method of dengue virus were established. Ten cases of positive cases of dengue virus were detected by ELISA, and the clinical serum samples of 1 to 2 days of fever were collected for quantitative PCR. Results: Real-time fluorescence PCR was positive for dengue virus type 1-4, and the probe and primer were universal probes and primers for dengue virus detection. Real-time fluorescence PCR detection of 10 clinical serum, 6 were positive, the positive rate was 60%. Conclusion: The real-time PCR method is a rapid, specific and sensitive method for detecting dengue virus. It is suitable for early clinical diagnosis of dengue fever and has good social and economic benefits.