tetano
Editor, Senior Moderator
J Virol Methods. 2017 Dec 15. pii: S0166-0934(17)30411-1. doi: 10.1016/j.jviromet.2017.12.005. [Epub ahead of print]
[h=1]Evaluation of clinical applicability of reverse transcription loop-mediated isothermal amplification assay for detection and subtyping of Influenza A viruses.[/h] Sharma V[SUP]1[/SUP], Chaudhry D[SUP]2[/SUP], Kaushik S[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] Influenza A viruses (IAVs) have always remain a serious concern for the global economic and public health. A rapid, specific and sensitive detection method is always needed to control the influenza in its early stages by timely intervention of therapy and early clinical management.
[h=4]OBJECTIVES:[/h] To develop RT-LAMP assays for detection of influenza A viruses, their further subtyping into seasonal (H1N1, H3N2) and novel pandemic H1N1 viruses and to evaluate clinical applicability of optimized RT-LAMP assays on patients' samples.
[h=4]STUDY DESIGN:[/h] In this study, we optimized RT-LAMP assay to detect IAVs by using primers against matrix gene and subtyping of IAVs was done by using primers against hemagglutinin gene. Optimized RT-LAMP assays were applied on clinical samples from patients having influenza like illness and results were compared with conventional one-step RT-PCR and real-time RT-PCR.
[h=4]RESULTS:[/h] RT-LAMP assays successfully detected and differentiated IAVs into H1N1, H3N2 and pdm09/H1N1 subtypes. One hundred and sixty seven clinical swab samples from influenza suspected patients were taken and tested with RT-LAMP assay, detecting 30 (17.9%) samples positive for Influenza A virus. Out of 30 samples, 21, 7 and 2 were found positive for pdm09/H1N1, H3N2 and seasonal H1 respectively. Conventional one-step RT-PCR detected a total of 27 (16.2%) samples for influenza A and further subtyping showed 20 and 7 samples positive for pdm09/H1N1 and H3N2 virus respectively whereas none was found positive for seasonal H1N1. RT-LAMP assay demonstrated higher sensitivity (93.8%) than conventional RT-PCR (84.4%) for influenza A viruses detection in clinical samples.
[h=4]CONCLUSIONS:[/h] RT-LAMP assay was rapid, sensitive, specific and cost effective method for detection of influenza A viruses than conventional one-step RT-PCR and it can serve as a good alternate for diagnosis and surveillance studies during influenza outbreaks in resource-limited setups of developing countries.
Copyright ? 2017. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Influenza A viruses; Molecular detection; Pandemic influenza; RT-LAMP
PMID: 29253497 DOI: 10.1016/j.jviromet.2017.12.005
[h=1]Evaluation of clinical applicability of reverse transcription loop-mediated isothermal amplification assay for detection and subtyping of Influenza A viruses.[/h] Sharma V[SUP]1[/SUP], Chaudhry D[SUP]2[/SUP], Kaushik S[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] Influenza A viruses (IAVs) have always remain a serious concern for the global economic and public health. A rapid, specific and sensitive detection method is always needed to control the influenza in its early stages by timely intervention of therapy and early clinical management.
[h=4]OBJECTIVES:[/h] To develop RT-LAMP assays for detection of influenza A viruses, their further subtyping into seasonal (H1N1, H3N2) and novel pandemic H1N1 viruses and to evaluate clinical applicability of optimized RT-LAMP assays on patients' samples.
[h=4]STUDY DESIGN:[/h] In this study, we optimized RT-LAMP assay to detect IAVs by using primers against matrix gene and subtyping of IAVs was done by using primers against hemagglutinin gene. Optimized RT-LAMP assays were applied on clinical samples from patients having influenza like illness and results were compared with conventional one-step RT-PCR and real-time RT-PCR.
[h=4]RESULTS:[/h] RT-LAMP assays successfully detected and differentiated IAVs into H1N1, H3N2 and pdm09/H1N1 subtypes. One hundred and sixty seven clinical swab samples from influenza suspected patients were taken and tested with RT-LAMP assay, detecting 30 (17.9%) samples positive for Influenza A virus. Out of 30 samples, 21, 7 and 2 were found positive for pdm09/H1N1, H3N2 and seasonal H1 respectively. Conventional one-step RT-PCR detected a total of 27 (16.2%) samples for influenza A and further subtyping showed 20 and 7 samples positive for pdm09/H1N1 and H3N2 virus respectively whereas none was found positive for seasonal H1N1. RT-LAMP assay demonstrated higher sensitivity (93.8%) than conventional RT-PCR (84.4%) for influenza A viruses detection in clinical samples.
[h=4]CONCLUSIONS:[/h] RT-LAMP assay was rapid, sensitive, specific and cost effective method for detection of influenza A viruses than conventional one-step RT-PCR and it can serve as a good alternate for diagnosis and surveillance studies during influenza outbreaks in resource-limited setups of developing countries.
Copyright ? 2017. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Influenza A viruses; Molecular detection; Pandemic influenza; RT-LAMP
PMID: 29253497 DOI: 10.1016/j.jviromet.2017.12.005