逆转录聚合酶链反应检测Ⅱ型登革病毒基因  

Detection of Dengue Virus 2 Genome by Reverse Transcription—Polymerase Chain Reaction

作　　者：王飞[1] 李刚[1] 郭日波[1] 柯伟民[1] 罗慧容 疗育煌 

机构地区：[1]中山医科大学传染病学教研室,广州510630 [2]广州市卫生防疫站

出　　处：《中国病毒学》1993年第2期138-141,共4页Virologica Sinica

摘　　要：本文应用逆转录聚合酶链反应(RT-PCR)方法检测Ⅱ型登革病毒基因。所设计引物在E基因区,引物1位于碱基序列的1566—1586,引物2位于1437—1458。引物在黄病毒属中为Ⅱ型登革病毒特有,它是Ⅱ型各株保守区。反应产物为150bp,内含一个HindⅢ酶位点,酶切后有111bP和39bp两个片段。使用本法对一系列稀释的培养液进行检测,可检出少至5TCID_(50)的病毒RNA。此外还检测了10份经病毒分离与免疫萤光分型证实为Ⅱ型登革热病人的血清和14份疑似Ⅱ型登革热病人但病毒分离阴性的急性期血清。证明本法敏感性明显高于病毒分离。Reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of dengue virus 2 genome. Two primers specific for dengue virus 2 and conservative between geographical variants bracketed a 150 nucleotide base in the E gene, corresponding to bases 1437—1586 in the dengue 2 genomic RNA sequence. After digested with Hind Ⅲ, the products of the reaction was divided into two fragments, 111bp and 39bp, respectively. The amplified products and digested fragments were shown in the ethidium bromide stained 2% agrose gel. RT-PCR can detect dengue viral RNA from at least 5 TCID_(50) virus. It is more sensitive and rapid than viral culture, which was justified by 24 sera.

关 键 词：登革病毒 逆转录 聚合酶链反应 

分 类 号：R373.33[医药卫生—病原生物学]