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KMID : 0357319940290010079
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Journal of the Korean Society for Microbiology
1994 Volume.29 No. 1 p.79 ~ p.88
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Detection and Serotypic Differentiation of Hantaviruses Using Reverse Transcriptase-Polymerase Chain Reaction(RT-PCR)
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Abstract
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RT-PCR(reverse transcriptase-polymerase chain reaction) was used for detection and serotypic differentiation of hantaviruses. To establish the specific primers to each serotype of hantaviruses, the most heterologous nucleotide
sequences(appoximatly
20
mers size) among M segments of Hantaan, Seoul, Puumala and Prospect Hill viruses were selected and built as candidates for the specific primers. The primer sets which amplify specifically the responsible serotypes were able to be established
successfully and the serotypes of hantavirus were also able to be differentiated by the primers. For the purpose of detection of hantavirus from clinical specimens, the universal primers which amplify all of hantaviruses could be built from the
moet
homologous sites selected by analyzing nucleotide sequences of S segments of hantavius serotypes. The analysis of restriction profiles of the DNA fragments amplified by the universal primer set was done for serotypic differentiation by using
several
restriction enzymes. The most typical restriction profiles could be obtained by the enzyme, Alu I. Hantaan viral RNA could be amplified from the several cases among the specimens from the patients of hemorrhagic fever with renal syndrome(HFRS) by
using
the specific primer set built. The previous attempt to isolate the virus from the specimens by traditional methods have been failed, therefore, this suggests that RT-PCR is considered as a useful and sensitive method for direct detection of
hantaviruses
from the clinical HRRS specimens.
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KEYWORD
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