Cargando…

Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus

The recent emergence of the swine-origin influenza A H1N1 virus (S-OIV) poses a serious global health threat. Rapid detection and differentiation of S-OIV from seasonal influenza is crucial for patient management and control of the epidemics. A one-step, single-tube accelerated and quantitative S-OI...

Descripción completa

Detalles Bibliográficos
Autores principales: Parida, Manmohan, Shukla, Jyoti, Sharma, Shashi, Ranghia Santhosh, Sanna, Ravi, Vasanthapuram, Mani, Reeta, Thomas, Maria, Khare, Shashi, Rai, Arvind, Kant Ratho, Radha, Pujari, Sujit, Mishra, Bijayanti, Lakshmana Rao, Putcha Venkata, Vijayaraghavan, Rajagopalan
Formato: Texto
Lenguaje:English
Publicado: American Society for Investigative Pathology 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3069812/
https://www.ncbi.nlm.nih.gov/pubmed/21227400
http://dx.doi.org/10.1016/j.jmoldx.2010.11.003
_version_ 1782201359223750656
author Parida, Manmohan
Shukla, Jyoti
Sharma, Shashi
Ranghia Santhosh, Sanna
Ravi, Vasanthapuram
Mani, Reeta
Thomas, Maria
Khare, Shashi
Rai, Arvind
Kant Ratho, Radha
Pujari, Sujit
Mishra, Bijayanti
Lakshmana Rao, Putcha Venkata
Vijayaraghavan, Rajagopalan
author_facet Parida, Manmohan
Shukla, Jyoti
Sharma, Shashi
Ranghia Santhosh, Sanna
Ravi, Vasanthapuram
Mani, Reeta
Thomas, Maria
Khare, Shashi
Rai, Arvind
Kant Ratho, Radha
Pujari, Sujit
Mishra, Bijayanti
Lakshmana Rao, Putcha Venkata
Vijayaraghavan, Rajagopalan
author_sort Parida, Manmohan
collection PubMed
description The recent emergence of the swine-origin influenza A H1N1 virus (S-OIV) poses a serious global health threat. Rapid detection and differentiation of S-OIV from seasonal influenza is crucial for patient management and control of the epidemics. A one-step, single-tube accelerated and quantitative S-OIV-specific H1 reverse transcription loop-mediated isothermal amplification (RTLAMP) assay for clinical diagnosis of S-OIV by targeting the H1 gene is reported in this article. A comparative evaluation of the H1-specific RTLAMP assay vis-à-vis the World Health Organization-approved real-time polymerase chain reaction (RTPCR), involving 239 acute-phase throat swab samples, demonstrated exceptionally higher sensitivity by picking up all of the 116 H1N1-positive cases and 36 additional positive cases among the negatives that were sequence-confirmed as S-OIV H1N1. None of the real-time RTPCR-positive samples were missed by the RTLAMP system. The comparative analysis revealed that S-OIV RTLAMP was up to tenfold more sensitive than the World Health Organization real-time RTPCR; it had a detection limit of 0.1 tissue culture infectious dosage of (50)/ml. One of the most attractive features of this isothermal gene amplification assay is that it seems to have an advantage in monitoring gene amplification by means of SYBR Green I dye-mediated naked-eye visualization within 30 minutes compared to 2 to 3 hours for a real-time reverse transcription polymerase chain reaction. This suggests that the RTLAMP assay is a valuable tool for rapid, real-time detection and quantification of S-OIV in acute-phase throat swab samples without requiring sophisticated equipment.
format Text
id pubmed-3069812
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher American Society for Investigative Pathology
record_format MEDLINE/PubMed
spelling pubmed-30698122012-01-01 Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus Parida, Manmohan Shukla, Jyoti Sharma, Shashi Ranghia Santhosh, Sanna Ravi, Vasanthapuram Mani, Reeta Thomas, Maria Khare, Shashi Rai, Arvind Kant Ratho, Radha Pujari, Sujit Mishra, Bijayanti Lakshmana Rao, Putcha Venkata Vijayaraghavan, Rajagopalan J Mol Diagn Regular Article The recent emergence of the swine-origin influenza A H1N1 virus (S-OIV) poses a serious global health threat. Rapid detection and differentiation of S-OIV from seasonal influenza is crucial for patient management and control of the epidemics. A one-step, single-tube accelerated and quantitative S-OIV-specific H1 reverse transcription loop-mediated isothermal amplification (RTLAMP) assay for clinical diagnosis of S-OIV by targeting the H1 gene is reported in this article. A comparative evaluation of the H1-specific RTLAMP assay vis-à-vis the World Health Organization-approved real-time polymerase chain reaction (RTPCR), involving 239 acute-phase throat swab samples, demonstrated exceptionally higher sensitivity by picking up all of the 116 H1N1-positive cases and 36 additional positive cases among the negatives that were sequence-confirmed as S-OIV H1N1. None of the real-time RTPCR-positive samples were missed by the RTLAMP system. The comparative analysis revealed that S-OIV RTLAMP was up to tenfold more sensitive than the World Health Organization real-time RTPCR; it had a detection limit of 0.1 tissue culture infectious dosage of (50)/ml. One of the most attractive features of this isothermal gene amplification assay is that it seems to have an advantage in monitoring gene amplification by means of SYBR Green I dye-mediated naked-eye visualization within 30 minutes compared to 2 to 3 hours for a real-time reverse transcription polymerase chain reaction. This suggests that the RTLAMP assay is a valuable tool for rapid, real-time detection and quantification of S-OIV in acute-phase throat swab samples without requiring sophisticated equipment. American Society for Investigative Pathology 2011-01 /pmc/articles/PMC3069812/ /pubmed/21227400 http://dx.doi.org/10.1016/j.jmoldx.2010.11.003 Text en © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
spellingShingle Regular Article
Parida, Manmohan
Shukla, Jyoti
Sharma, Shashi
Ranghia Santhosh, Sanna
Ravi, Vasanthapuram
Mani, Reeta
Thomas, Maria
Khare, Shashi
Rai, Arvind
Kant Ratho, Radha
Pujari, Sujit
Mishra, Bijayanti
Lakshmana Rao, Putcha Venkata
Vijayaraghavan, Rajagopalan
Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus
title Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus
title_full Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus
title_fullStr Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus
title_full_unstemmed Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus
title_short Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus
title_sort development and evaluation of reverse transcription loop-mediated isothermal amplification assay for rapid and real-time detection of the swine-origin influenza a h1n1 virus
topic Regular Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3069812/
https://www.ncbi.nlm.nih.gov/pubmed/21227400
http://dx.doi.org/10.1016/j.jmoldx.2010.11.003
work_keys_str_mv AT paridamanmohan developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT shuklajyoti developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT sharmashashi developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT ranghiasanthoshsanna developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT ravivasanthapuram developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT manireeta developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT thomasmaria developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT khareshashi developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT raiarvind developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT kantrathoradha developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT pujarisujit developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT mishrabijayanti developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT lakshmanaraoputchavenkata developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus
AT vijayaraghavanrajagopalan developmentandevaluationofreversetranscriptionloopmediatedisothermalamplificationassayforrapidandrealtimedetectionoftheswineorigininfluenzaah1n1virus