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Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses
BACKGROUND: Dengue is emerging as a major public health concern in many parts of the world. The development of a one-step, single tube, rapid, and multiplex reverse transcription polymerase chain reaction (M-RT-PCR) for simultaneous detection and typing of dengue virus using serotype specific primer...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267776/ https://www.ncbi.nlm.nih.gov/pubmed/18234069 http://dx.doi.org/10.1186/1743-422X-5-20 |
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author | Saxena, Parag Dash, Paban Kumar Santhosh, SR Shrivastava, Ambuj Parida, Manmohan Rao, PV Lakshmana |
author_facet | Saxena, Parag Dash, Paban Kumar Santhosh, SR Shrivastava, Ambuj Parida, Manmohan Rao, PV Lakshmana |
author_sort | Saxena, Parag |
collection | PubMed |
description | BACKGROUND: Dengue is emerging as a major public health concern in many parts of the world. The development of a one-step, single tube, rapid, and multiplex reverse transcription polymerase chain reaction (M-RT-PCR) for simultaneous detection and typing of dengue virus using serotype specific primers during acute phase of illness is reported. RESULTS: An optimal assay condition with zero background was established having no cross-reaction with closely related members of flavivirus (Japanese encephalitis, West Nile, Yellow fever) and alphavirus (Chikungunya). The feasibility of M-RT-PCR assay for clinical diagnosis was validated with 620 acute phase dengue patient sera samples of recent epidemics in India. The comparative evaluation vis a vis conventional virus isolation revealed higher sensitivity. None of the forty healthy serum samples screened in the present study revealed any amplification, thereby establishing specificity of the reported assay for dengue virus only. CONCLUSION: These findings clearly suggested that M-RT-PCR assay reported in the present study is the rapid and cost-effective method for simultaneous detection as well as typing of the dengue virus in acute phase patient serum samples. Thus, the M-RT-PCR assay developed in this study will serve as a very useful tool for rapid diagnosis and typing of dengue infections in endemic areas. |
format | Text |
id | pubmed-2267776 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-22677762008-03-15 Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses Saxena, Parag Dash, Paban Kumar Santhosh, SR Shrivastava, Ambuj Parida, Manmohan Rao, PV Lakshmana Virol J Methodology BACKGROUND: Dengue is emerging as a major public health concern in many parts of the world. The development of a one-step, single tube, rapid, and multiplex reverse transcription polymerase chain reaction (M-RT-PCR) for simultaneous detection and typing of dengue virus using serotype specific primers during acute phase of illness is reported. RESULTS: An optimal assay condition with zero background was established having no cross-reaction with closely related members of flavivirus (Japanese encephalitis, West Nile, Yellow fever) and alphavirus (Chikungunya). The feasibility of M-RT-PCR assay for clinical diagnosis was validated with 620 acute phase dengue patient sera samples of recent epidemics in India. The comparative evaluation vis a vis conventional virus isolation revealed higher sensitivity. None of the forty healthy serum samples screened in the present study revealed any amplification, thereby establishing specificity of the reported assay for dengue virus only. CONCLUSION: These findings clearly suggested that M-RT-PCR assay reported in the present study is the rapid and cost-effective method for simultaneous detection as well as typing of the dengue virus in acute phase patient serum samples. Thus, the M-RT-PCR assay developed in this study will serve as a very useful tool for rapid diagnosis and typing of dengue infections in endemic areas. BioMed Central 2008-01-30 /pmc/articles/PMC2267776/ /pubmed/18234069 http://dx.doi.org/10.1186/1743-422X-5-20 Text en Copyright © 2008 Saxena et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Saxena, Parag Dash, Paban Kumar Santhosh, SR Shrivastava, Ambuj Parida, Manmohan Rao, PV Lakshmana Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses |
title | Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses |
title_full | Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses |
title_fullStr | Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses |
title_full_unstemmed | Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses |
title_short | Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses |
title_sort | development and evaluation of one step single tube multiplex rt-pcr for rapid detection and typing of dengue viruses |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267776/ https://www.ncbi.nlm.nih.gov/pubmed/18234069 http://dx.doi.org/10.1186/1743-422X-5-20 |
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