The Study on the Genetic Varition of Type 1 dengue Virus Isola- tes in Taiwan, 1987-1992.

• Main Authors: Yi-Ching Dong, 董宜青
• Other Authors: Shui-Fung Chang
• Format: Others
• Language: zh-TW
• Published: 1993
• Online Access: http://ndltd.ncl.edu.tw/handle/72960335051782382124

碩士 === 高雄醫學院 === 醫學研究所 === 81 === Reverse transcriptase - polymerase chain reaction (RT-PCR) was used for the detection and typing of dengue virus. Briefly cDNA copy of a portion of the viral genome was synthesized in a reverse transcriptase reaction in the presence of complementary primer and then PCR was carried out for 35 cycles of heat denaturation, annealing, and primer extention with the addition of sense. The product of RT-PCR was visualized as bands of appropriate size on ethidium bromide-stained agarose gels. The sensitivity of RT-PCR is 500 PFU (plaque forming unit). The crossreation test revealed that each primer pair anneal its own templete specifically. Thus RT-PCR provided a rapid and accurate method for the detection and typing of dengue virus. The genetic variation of type 1 dengue virus was studied using 9 strains, included 3 imported strains, isolated in Taiwan for 1987 to 1992. By camparison of the neucleotide sequence(E\NS1 junction, 490bp, nucleotide NO.2229 to 2718) of these strains, we found that Taiwan strains shared geographic similarity and the difference among these strains was only 1.53%. But the differenceamong imported strains and other strains was 2.87%. Since most for the point mutation of nucleotide sequence occurred in the third position of the codon, the amino acid sequence differed in 1 or 2 amino acid among the isolates.