TaqMan MGB实时荧光定量PCR检测登革Ⅰ型病毒

目的检测发热患者血清中的登革病毒含量,并判定其型别。方法在登革病毒的E基因区设计一对引物和一条MGB探针,建立TaqMan MGB实时荧光定量PCR体系;从10份疑似登革热患者血清中提取病毒RNA,逆转录成cDNA,采用TaqMan MGB实时荧光定量PCR技术检测登革病毒及其型别。结果10份血清样本中,有9例出现阳性扩增曲线,病毒含量在103-105拷贝/ml,且扩增产物均出现大约100 bp的扩增带。结论2006年在广州市流行的登革热是由登革Ⅰ型病毒引起,TaqMan MGB实时荧光定量PCR检测登革病毒感染具有快速、敏感的特点,为登革热的早期诊断提供了可能。

Dengue Type Ⅰ Virus Isolated in Guangzhou Detected by Using TaqMan MGB Real-time PCR

OBJECTIVE To detect serotype of dengue virus in sera from patients with fever. METHODS A pair of degenerated primers and one MGB probe were designed targeting the conserved region at the E gene of dengue type 1 virus. TaqMan MGB real-time PCR assay was developed with plasmid including E gene of dengue type 1 virus as standard sample. The sera of 10 patients with fever were used to extract RNA, and convert into eDNA. Then eDNA were detected by TaqMan MGB real-time PCR assay and the amplified products were analyzed at the same time. RESULTS The sera of 9 patients from 10 samples were observed to generate a fluorescent signal, and about 100 bp fragment was obtained simultaneously. CONCLUSIONS Dengue fever on 2006 in Guangzhou is caused by the dengue type 1 virus. TaqMan MGB real-time PCR assay is rapid and sensitive to detect dengue virus infections.