Li G
Department of Infectious Disease, Sun Yat-sen University of Medical Sciences, Guangzhou.
Zhonghua Yi Xue Za Zhi. 1993 Oct;73(10):605-8, 638.
RT-PCR was developed for the amplification of partial E genome fragment from four dengue serotypes. Of the six oligonucleotide primers designed one was shared by dengue virus type 1 and type 2, and one by type 3 and type 4. Each of the other four primers was type specific. Amplified products with 240, 150, 333 and 421 bp, respectively were identified by electrophoresis on 2% agarose gel and digestion with restriction enzyme Hind III. RT-PCR can detect dengue viral RNA from at least 5TCID50 virus, which was confirmed by detection of serial dilutions of culture supernatants. RT-PCR was also applied to serum samples from 60 acute patients. The findings showed that RT-PCR was of the same specificity as isolation of virus, followed by indirect fluorescent antibody tests. RT-PCR was more sensitive than cell culture and can be used for the rapid diagnosis and serotype identification of dengue virus infections.
逆转录聚合酶链反应（RT-PCR）用于扩增登革热四种血清型的部分E基因组片段。在设计的六种寡核苷酸引物中，有一种为登革热1型和2型所共有，另一种为3型和4型所共有。其余四种引物均为各血清型所特有。通过2%琼脂糖凝胶电泳和用限制性内切酶Hind III消化，分别鉴定出了长度为240、150、333和421 bp的扩增产物。RT-PCR能够从至少5个半数组织培养感染剂量（TCID50）的病毒中检测到登革病毒RNA，这通过对培养上清液的系列稀释检测得到了证实。RT-PCR还应用于60例急性患者的血清样本。结果表明，RT-PCR与病毒分离具有相同的特异性，随后进行间接荧光抗体试验。RT-PCR比细胞培养更敏感，可用于登革病毒感染的快速诊断和血清型鉴定。
