逆转录聚合酶链反应检测Ⅲ型登革病毒的敏感性和特异性研究

STUDIES ON SENSITIVITY AND SPECIFICITY OF REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION IN DETECTION OF DENGUE 3 VIRUS

本文应用逆转录聚合酶链反应(RT-PCR)方法,检测Ⅲ型登革病毒基因。所设计引物在E基因组,引物2位于核苷酸序列的1139～1158位,引物1位于1453～1471位,反应产物为333bp,内含1个HindⅢ限制性内切酶位点,酶切后有128 bp和199 bp两个片段。产物在含溴化乙锭的2％琼脂糖中电泳。采用RT-PCR法可测出少至5个半数组织细胞培养感染量(TCID_so)的病毒RNA。

Reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of dengue virus 3 genome. Two primers bracketed a 333-nucleotide base in the E gene, corresponding to bases 1139-1471 in the dengue 3 genomic RNA sequence. After digested with Hind Ⅲ, the product of the reaction was divided into two fragments, 128bp and 199bp, respectively. The amplified product and digested fragments were shown in the ethidium bromide stained 2% agrose gel. This technique can detect dengue viral RNA in the level of 5 TCID50 virus.