Comparative Analysis of Molecular Methods for Detection of Influenza Viruses
Vikrant Sharma, Samander Kaushik
Microbiology Research Journal International · pp. 1–10 · Published 4 Oct 2016
10.9734/BMRJ/2016/28858Abstract
Aims: Influenza is a serious threat to human population worldwide therefore continuous surveillance is required to update influenza seasonal vaccines. A rapid, sensitive, specific and cost effective diagnostic method will be much helpful for patient management in the present scenario. Present study is conceptualized for detection of influenza viruses by molecular methods and compare with ‘gold standard’ virus isolation. Study Design: Standard strains of Influenza virus were used to standardize the molecular diagnostic assays and results were then compared with virus isolation. Place and Duration of Study: Centre for Biotechnology, Maharshi Dayanand University, Rohtak, Haryana, India, between December 2015 and April 2016. Methodology: Standard strains of Influenza A and B virus were used for influenza virus isolation using virus culturing in MDCK (Madin-Darby Canine Kidney) cell line by following standard tissue culture procedure. Isolated viruses were detected by Hemagglutination assay (HA) and typed by Hemagglutination inhibition assay (HI). Conventional one step RT-PCR, Taqman real time RT-PCR and RT-LAMP (Reverse transcription loop mediated isothermal amplification) were standardized on RNA extracted from standard strains. Sensitivity and specificity of these molecular methods were compared with each other as well as with virus culture (gold standard). Results: Both influenza A and B virus strains were cultured in MDCK cells and produced cytopathic effect during virus culture. Conventional RT-PCR and real time RT-PCR detected both type of Influenza viruses. RT-LAMP also successfully detected and typed influenza viruses. RT-LAMP proved to be more rapid than other two molecular assays. Conclusion: Molecular diagnostic methods are useful in detection and typing of Influenza viruses and these methods provide results in short period of time when compared with traditional virus culture methods. RT-LAMP is rapid, sensitive, specific and cost effective method for influenza virus detection and subtyping.
References (30)
- 1 Loop-mediated isothermal amplification of DNA [DOI]
- 2 Global Patterns of Influenza A Virus in Wild Birds [DOI]
- 3 Global Epidemiology of Influenza: Past and Present [DOI]
- 4 Efficacy and Safety of the Neuraminidase Inhibitor Zanamivir in the Treatment of Influenzavirus Infections [DOI]
- 5 THE QUANTITATIVE DETERMINATION OF INFLUENZA VIRUS AND ANTIBODIES BY MEANS OF RED CELL AGGLUTINATION [DOI]
- 6 Simultaneous Detection of Influenza Viruses A and B Using Real-Time Quantitative PCR [DOI]
- 7 Poor Clinical Sensitivity of Rapid Antigen Test for Influenza A Pandemic (H1N1) 2009 Virus [DOI]
- 8 Development of H5-RT-LAMP (loop-mediated isothermal amplification) system for rapid diagnosis of H5 avian influenza virus infection [DOI]
- 9 Cocirculation of Two Distinct Genetic and Antigenic Lineages of Proposed Influenza D Virus in Cattle [DOI]
- 10 Serological evidence for the presence of influenza D virus in small ruminants [DOI]
- 11 An Open Receptor-Binding Cavity of Hemagglutinin-Esterase-Fusion Glycoprotein from Newly-Identified Influenza D Virus: Basis for Its Broad Cell Tropism [DOI]
- 12 Typing and Subtyping Influenza Virus Using DNA Microarrays and Multiplex Reverse Transcriptase PCR [DOI]
- 13 Detection of Human Influenza A Viruses by Loop-Mediated Isothermal Amplification [DOI]
- 14 Loop-Mediated Isothermal Amplification for Influenza A (H5N1) Virus [DOI]
- 15 Development of a real-time RT-PCR for the detection of Swine-lineage Influenza A (H1N1) virus infections [DOI]
- 16 Development of a Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Detection of Pandemic (H1N1) 2009 Virus as a Novel Molecular Method for Diagnosis of Pandemic Influenza in Resource-Limited Settings [DOI]
- 17 Yield of Sputum for Viral Detection by Reverse Transcriptase PCR in Adults Hospitalized with Respiratory Illness [DOI]
- 18 Identification of A/H5N1 Influenza Viruses Using a Single Gene Diagnostic Microarray [DOI]
- 19 Currently used nucleic acid amplification tests for the detection of viruses and atypicals in acute respiratory infections [DOI]
- 20 Performance of a Rapid Influenza Test in Children During the H1N1 2009 Influenza A Outbreak [DOI]
- 21 Diagnostic Accuracy of a Rapid Influenza Test for Pandemic Influenza A H1N1 [DOI]
- 22 A Combination of Serological Assays to Detect Human Antibodies to the Avian Influenza A H7N9 Virus [DOI]
- 23 Comparison of enzyme-linked immunosorbent assay, indirect immunofluorescence assay, and virus isolation for detection of respiratory viruses in nasopharyngeal secretions [DOI]
- 24 Type- and subtype-specific detection of influenza viruses in clinical specimens by rapid culture assay [DOI]
- 25 Direct detection of influenza virus antigen in nasopharyngeal specimens by direct enzyme immunoassay in comparison with quantitating virus shedding [DOI]
- 26 Development of a reverse transcription loop-mediated isothermal amplification method for the rapid detection of avian influenza virus subtype H7 [DOI]
- 27 Reverse-transcription, loop-mediated isothermal amplification assay for the sensitive and rapid detection of H10 subtype avian influenza viruses [DOI]
- 28 Comparison of RT-PCR with other diagnostic assays for rapid detection of influenza viruses [DOI]
- 29 Prospective comparative study of culture specimens and methods in diagnosing influenza in adults [DOI]
- 30 Update on Emerging Infections from the Centers for Disease Control and Prevention [DOI]
Cited by 13
Vikrant Sharma, Deepak Kumar, Divya Dhull, Sulochana Kaushik, Jaya Parkash Yadav, Samander Kaushik · International Blood Research & Reviews · 2017
F. Contreras, Katherine Rivero, J. Rivas-Pardo · International Journal of Molecular Sciences · 2025
Sanjit Boora, Anish Khan, Kumari Soniya · Research Journal of Pharmacy and Technology · 2023
Hakimeh Baghaei Daemi, M. F. Kulyar, Xinlin He · Viruses · 2021
B. Supraja, Vardarajalu, P. Karkuzhali · 2021
Vikrant Sharma, D. Chaudhry, S. Kaushik · Journal of Virological Methods · 2017
Vikrant Sharma, Deepak Kumar, Divya Dhull · 2017
J. Olaitan, O. B. Shittu · 2014
Bhoj R Singh, Akhilesh Kumar, Ravi Kant Agrawal · Acta Scientific Microbiology · 2022
Lucien Jacky, Dominic Yurk, John Alvarado · Analytical Chemistry · 2021
Showing 10 of 13 known citations — external sources report more than can currently be individually listed.
Related research
- Analysis of Influenza A/Beijing/353/89 Virus Matrix Gene Sequence and Identification of Replaceable Amino Acids as Natural Mutations in M2 Protein — shares topic coverage
- Pathogenesis of the Influenza Virus in Diabetes Model Mice — shares topic coverage
- Effective Inhibition of H5N1 Influenza Virus Infectivity using Safe and Calculated Low Power Generated from Life Restoration device under Different Conditions — shares topic coverage
- Evaluating the Effect of Amine-geldanamycin Hybrids on Antiviral Activity against Influenza Virus — shares topic coverage
Article metrics
Real usage data collected on this platform.
4
Page views
0
PDF downloads
0
Outbound clicks
13
Citations
Views over time
Views by country
Approximate, from request IP at view time — not citizenship or institution. Countries with fewer than 5 views are grouped as "Other".
Traffic sources
Referring site, by host.
Views and downloads exclude known bots/crawlers. Citations combines this platform's own DOI-resolved index with each external source's own reported total — see Cited by above for individually listed citing works. Last refreshed 0 seconds ago.