Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR

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 pati...

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Main Authors: Nastiti Wijayanti, Hera Nirwati, Tri Wibawa, Aris Haryanto, S. Sutaryo
Format: Article
Language:English
Published: Universitas Gadjah Mada, Yogyakarta 2006-12-01
Series:Indonesian Journal of Biotechnology
Online Access:https://jurnal.ugm.ac.id/ijbiotech/article/view/7569
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author Nastiti Wijayanti
Hera Nirwati
Tri Wibawa
Aris Haryanto
S. Sutaryo
author_facet Nastiti Wijayanti
Hera Nirwati
Tri Wibawa
Aris Haryanto
S. Sutaryo
author_sort Nastiti Wijayanti
collection DOAJ
description 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
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spelling doaj.art-09e5346c7c8544848df59b278115ffb92022-12-21T20:48:02ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412006-12-0111210.22146/ijbiotech.75696370Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCRNastiti WijayantiHera NirwatiTri WibawaAris HaryantoS. Sutaryoworld. 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 PCRhttps://jurnal.ugm.ac.id/ijbiotech/article/view/7569
spellingShingle Nastiti Wijayanti
Hera Nirwati
Tri Wibawa
Aris Haryanto
S. Sutaryo
Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
Indonesian Journal of Biotechnology
title Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
title_full Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
title_fullStr Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
title_full_unstemmed Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
title_short Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR
title_sort rapid detection and molecular typing of dengue virus by using multiplex nested rt pcr
url https://jurnal.ugm.ac.id/ijbiotech/article/view/7569
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