Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus
The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe b...
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th-mahidol.215532018-07-24T10:48:46Z Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus Wasun Chantratita Wiroj Pongtanapisit Wantanich Piroj Chutatip Srichunrasmi Somying Seesuai Mahidol University Medicine The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and a real-time assay with virus-specific molecular beacon probes (NASBA-Beacon assay). The analytical sensitivity or reproducibility of the test results among those three assays was compared. All assays yielded results by detecting SARS coronavirus targeting the BNI-1 region in less than 2 hours. RNA detection by all the formats was unaffected by the presence of human sputum. The limits of detection were at least 10 copies of input RNA for both RT-PCR formats (RT-PCR TaqMan and RT-PCR hybridization probe assays), while the NASBA-Beacon assay could detect as little as 1 copy per reaction, with high reproducibility of the coefficient of variation (CV) of <10. These results demonstrate that real-time NASBA provides a rapid and sensitive alternative to RT-PCR for the routine qualitative assay of sputum for SARS corona viral RNA detection. 2018-07-24T03:48:46Z 2018-07-24T03:48:46Z 2004-09-01 Article Southeast Asian Journal of Tropical Medicine and Public Health. Vol.35, No.3 (2004), 623-629 01251562 2-s2.0-8444228607 https://repository.li.mahidol.ac.th/handle/123456789/21553 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=8444228607&origin=inward |
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Medicine Wasun Chantratita Wiroj Pongtanapisit Wantanich Piroj Chutatip Srichunrasmi Somying Seesuai Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus |
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The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and a real-time assay with virus-specific molecular beacon probes (NASBA-Beacon assay). The analytical sensitivity or reproducibility of the test results among those three assays was compared. All assays yielded results by detecting SARS coronavirus targeting the BNI-1 region in less than 2 hours. RNA detection by all the formats was unaffected by the presence of human sputum. The limits of detection were at least 10 copies of input RNA for both RT-PCR formats (RT-PCR TaqMan and RT-PCR hybridization probe assays), while the NASBA-Beacon assay could detect as little as 1 copy per reaction, with high reproducibility of the coefficient of variation (CV) of <10. These results demonstrate that real-time NASBA provides a rapid and sensitive alternative to RT-PCR for the routine qualitative assay of sputum for SARS corona viral RNA detection. |
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Mahidol University |
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Mahidol University Wasun Chantratita Wiroj Pongtanapisit Wantanich Piroj Chutatip Srichunrasmi Somying Seesuai |
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Wasun Chantratita Wiroj Pongtanapisit Wantanich Piroj Chutatip Srichunrasmi Somying Seesuai |
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Wasun Chantratita |
title |
Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus |
title_short |
Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus |
title_full |
Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus |
title_fullStr |
Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus |
title_full_unstemmed |
Development and comparison of the real-time amplification based methods - NASBA-Beacon, RT-PCR TaqMan and RT-PCR hybridization probe assays - For the qualitative detection of sars coronavirus |
title_sort |
development and comparison of the real-time amplification based methods - nasba-beacon, rt-pcr taqman and rt-pcr hybridization probe assays - for the qualitative detection of sars coronavirus |
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2018 |
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https://repository.li.mahidol.ac.th/handle/123456789/21553 |
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