辽宁省锦州市首例输入性登革热病例的调查、快速检测方法与防控探讨

Survey, rapid detection method, prevention and control of first imported dengue fever of Jinzhou City in Liaoning Province

目的通过调查总结输入性登革热的防治经验,并建立Taqman MGB实时荧光PCR技术快速检测登革热病毒,更好地指导今后登革热防制工作。方法流行病学调查,利用Taqman MGB技术,根据登革病毒3’端非编码区一段序列,设计登革1~4型荧光PCR通用引物和探针,以DF病毒株作为标准,以乙脑毒株作阴性对照,建立荧光PCR检测DF的快速方法。同时给予1份ELISA法检测阳性的临床血清标本进行荧光PCR扩增。结果采用PCR引物对登革1~4型病毒进行扩增,与设计相符;而乙型脑炎病毒均无非特异性扩增条带,对引物的相关性实验结果表明引物之间不会因相互干扰而出现假阳性结果,1份疑似患者血标本RT-PCR扩增阳性率为83.3%(25/30),其核酸序列与登革1型病毒柬埔寨株以及中国1997、1999年流行株GD14/97、GD05/99同源性分别为97%。结论在今后的登革热防控工作中,我们应用RT-PCR检测方法时间短、灵敏性高、有较高的特异性,以此作为为DF的临床早期诊断方法;其次重点加强出国和归国人员的管理,开展登革热防治知识的宣传教育,建立和其他部门的登革热联防体制,加强基层医务人员登革热防治知识培训,提高基层医务人员登革热诊治水平,来达到控制登革热的目标。

Objective To summarize the experience of prevention and treatment of imported dengue fever by investigation and to establish Taqman MGB real-time fluorescence PCR to detect dengue virus rapidly so as to better guide dengue fever prevention and control work in the future. Methods The epidemiological investigation was carried out in the field. According to a highly conserved sequence of the non-coding region of dengue virus 3 ＇end, dengue 1.4 fluorescent PCR universal primer and Taqman MGB probe were designed, using Taqman MGB technique. The rapid detection method of real-time fluorescence PCR detection of dengue virus was established, with Dengue fever virus strain as the standard and JE as the control. The clinical serum sample of the one case of ELISA test positive were amplified by RT-PCR and fluorescent PCR amplification. Results The dengue virus type 1 to 4 were amplified using PCR primers,which was consistent with the design. However, there was no non-specific amplification band for Japanese encephalitis virus. The primer correlation experimental results showed that the false positive results would not occur due to mutual interference between the primers. The positive rate of RT-PCR amplification in one suspected patient＇s blood sample was 83.3%（25/30）. Its nucleotide sequence homology with dengue virus type 1 Cambodia, China GD14/97, GD05/99 in1997 and 1999 was 97%. Conclusion In the prevention and control of dengue fever in the future, RT-PCR is a rapid,sensitive and specific method, which can be used as an early clinical diagnostic method for dengue fever. Second, we will focus on strengthening the management of overseas personnel and returnees, develop the publicity and education of dengue fever prevention and treatment. We also establish the defense system of dengue fever with other departments,strengthen the training on dengue fever prevention and control among grassroots medical workers and improve the level of diagnosis and treatment of dengue fever among grassroots medical workers so as to achieve the goal of controlling dengue fever.