| Summary: | 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.
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