Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
https://doi.org/10.22146/ijbiotech.7569 Nastiti Wijayanti(1*), Hera Nirwati(2), Tri Wibawa(3), Aris Haryanto(4), S. Sutaryo(5)
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(*) Corresponding Author
Abstract
world. We have evaluated the combination of one-step RT-PCR and multiplex nested PCR assays for detecting
dengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primers
that conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirus
and there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,
the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virus
using multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.
Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,
whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assay
can be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiology
and also evolutionary studies.
Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR
dengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primers
that conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirus
and there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,
the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virus
using multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.
Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,
whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assay
can be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiology
and also evolutionary studies.
Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR
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PDF DOI: https://doi.org/10.22146/ijbiotech.7569
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