摘要
目的建立同时检测西尼罗病毒(WNV)、基孔肯雅病毒(CHIKV)的双重荧光定量PCR法,为临床疑似病例的诊断提供依据。方法分别针对WNV CAP基因、CHIKV E1基因保守区设计特异性引物和TaqMan探针,建立并优化双重荧光定量PCR反应体系,评价方法的特异性和灵敏度。结果建立的双重荧光定量PCR可同时检测WNV、CHIKV核酸,标准曲线相关系数(r)分别达0.999、0.998,灵敏度达10 copies/μL,具有良好的特异性。结论建立了同时检测WNV、CHIKV的双重荧光定量PCR法,但尚需临床进一步验证。
Objective To develop a duplex real-time quantitative PCR assay for simultaneous detection of West Nile virus and Chikungunya virus. Methods Two sets of primers and TaqMan probes were designed based on highly conserved CAP gene region of West Nile virus and E1 gene region of Chikungunya virus, and the reactive condition of duplex real-time PCR was optimized. The sensitivity and specificity of the assay were evaluated. Results The duplex real-time quantitative PCR assay showed excellent specificity in simultaneous detection of West Nile virus and Chikungunya virus. The sensitivities of the assay were 10 copies/μL for both the virus, and the correlation coefficient of the quantitative curve were 0. 999 and 0. 998 respectively. Conclusion The duplex fluorescent quantita- tive PCR assay developed in this study is sensitive and specific for simultaneous detection of West Nile virus and Chikungunya virus. The efficiency of the assay for detecting of clinical samples should be further evaluated.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2013年第6期412-414,共3页
Chinese Journal of Clinical Laboratory Science
基金
浙江省医学重点学科群建设项目(XKQ-009-003)
