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An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system
BACKGROUND: Dengue fever, a mosquito-borne disease, is caused by dengue virus (DENV) which includes four major serotypes (DENV-1, -2, -3, and -4). Some serotypes cause more severe diseases than the other; severe dengue is associated with secondary infections by a different serotype. Timely serotypin...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6433249/ https://www.ncbi.nlm.nih.gov/pubmed/30908535 http://dx.doi.org/10.1371/journal.pone.0214328 |
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author | Tsai, Jih-Jin Liu, Wei-Liang Lin, Ping-Chang Huang, Bo-Yi Tsai, Ching-Yi Chou, Pin-Hsing Lee, Fu-Chun Ping, Chia-Fong Lee, Pei-Yu Alison Liu, Li-Teh Chen, Chun-Hong |
author_facet | Tsai, Jih-Jin Liu, Wei-Liang Lin, Ping-Chang Huang, Bo-Yi Tsai, Ching-Yi Chou, Pin-Hsing Lee, Fu-Chun Ping, Chia-Fong Lee, Pei-Yu Alison Liu, Li-Teh Chen, Chun-Hong |
author_sort | Tsai, Jih-Jin |
collection | PubMed |
description | BACKGROUND: Dengue fever, a mosquito-borne disease, is caused by dengue virus (DENV) which includes four major serotypes (DENV-1, -2, -3, and -4). Some serotypes cause more severe diseases than the other; severe dengue is associated with secondary infections by a different serotype. Timely serotyping can provide early warning of dengue epidemics to improve management of patients and outbreaks. A mobile insulated isothermal PCR (iiPCR) system is available to allow molecular detection of pathogens near points of need. METHODOLOGY/PRINCIPLE FINDINGS: In this study, side-by-side comparison with the CDC DENV-1-4 Real Time RT-PCR (qRT-PCR) was performed to evaluate the performance of four singleplex DENV-1–4 serotyping reverse transcription-iiPCR (RT-iiPCR) reagents for DENV subtyping on the mobile PCR system. The four RT-iiPCRs did not react with Zika virus and chikungunya virus; tests with serial dilutions of the four DENV serotypes made in human serum showed they had detection endpoints comparable to those of the reference method, indicating great analytical sensitivity and specificity. Clinical performance of the RT-iiPCR reagents was evaluated by testing 40 serum samples each (around 20 target serotype-positive and 20 DENV-negative); all four reagents had high agreement (97.5–100%) with the reference qRT-PCR. Moreover, testing of mosquitoes separately infected experimentally with each serotype showed that the four reagents detected specifically their target DENV serotypes in mosquito. CONCLUSIONS/SIGNIFICANCE: With analytical and clinical performance comparable to the reference qRT-PCR assay, the four index RT-iiPCR reagents on the field-deployable PCR system can serve as a useful tool for DENV detection near points of needs. |
format | Online Article Text |
id | pubmed-6433249 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-64332492019-04-08 An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system Tsai, Jih-Jin Liu, Wei-Liang Lin, Ping-Chang Huang, Bo-Yi Tsai, Ching-Yi Chou, Pin-Hsing Lee, Fu-Chun Ping, Chia-Fong Lee, Pei-Yu Alison Liu, Li-Teh Chen, Chun-Hong PLoS One Research Article BACKGROUND: Dengue fever, a mosquito-borne disease, is caused by dengue virus (DENV) which includes four major serotypes (DENV-1, -2, -3, and -4). Some serotypes cause more severe diseases than the other; severe dengue is associated with secondary infections by a different serotype. Timely serotyping can provide early warning of dengue epidemics to improve management of patients and outbreaks. A mobile insulated isothermal PCR (iiPCR) system is available to allow molecular detection of pathogens near points of need. METHODOLOGY/PRINCIPLE FINDINGS: In this study, side-by-side comparison with the CDC DENV-1-4 Real Time RT-PCR (qRT-PCR) was performed to evaluate the performance of four singleplex DENV-1–4 serotyping reverse transcription-iiPCR (RT-iiPCR) reagents for DENV subtyping on the mobile PCR system. The four RT-iiPCRs did not react with Zika virus and chikungunya virus; tests with serial dilutions of the four DENV serotypes made in human serum showed they had detection endpoints comparable to those of the reference method, indicating great analytical sensitivity and specificity. Clinical performance of the RT-iiPCR reagents was evaluated by testing 40 serum samples each (around 20 target serotype-positive and 20 DENV-negative); all four reagents had high agreement (97.5–100%) with the reference qRT-PCR. Moreover, testing of mosquitoes separately infected experimentally with each serotype showed that the four reagents detected specifically their target DENV serotypes in mosquito. CONCLUSIONS/SIGNIFICANCE: With analytical and clinical performance comparable to the reference qRT-PCR assay, the four index RT-iiPCR reagents on the field-deployable PCR system can serve as a useful tool for DENV detection near points of needs. Public Library of Science 2019-03-25 /pmc/articles/PMC6433249/ /pubmed/30908535 http://dx.doi.org/10.1371/journal.pone.0214328 Text en © 2019 Tsai et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Tsai, Jih-Jin Liu, Wei-Liang Lin, Ping-Chang Huang, Bo-Yi Tsai, Ching-Yi Chou, Pin-Hsing Lee, Fu-Chun Ping, Chia-Fong Lee, Pei-Yu Alison Liu, Li-Teh Chen, Chun-Hong An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system |
title | An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system |
title_full | An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system |
title_fullStr | An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system |
title_full_unstemmed | An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system |
title_short | An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system |
title_sort | rt-pcr panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable pcr system |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6433249/ https://www.ncbi.nlm.nih.gov/pubmed/30908535 http://dx.doi.org/10.1371/journal.pone.0214328 |
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