A rapid method for the analysis of influenza virus genes: application to the reassortment of equine influenza virus genes.
Abstract: We describe a rapid method for genetic characterisation of influenza virus genes using reverse transcription and amplification by polymerase chain reaction (RT/PCR) of all virus segments simultaneously (multiplex RT/PCR) using primers based on the conserved terminal sequences. The product has been shown to be suitable for determination of partial nucleotide sequences which can be used to search nucleotide sequence databases and rapidly map the genetic origin of each segment. We illustrate the use of the method by analysing genetic reassortment in H7N7 equine influenza viruses.
Publication Date: 1994-06-01 PubMed ID: 7521550DOI: 10.1016/0168-1702(94)90087-6Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article introduces a quick method for analyzing the genes of influenza virus by using reverse transcription and polymerase chain reaction. This method was applied to examine gene reassortment in H7N7 equine influenza viruses.
Method for Characterizing Influenza Virus Genes
- The researchers describe a method for fast genetic characterization of influenza virus genes. This technique involves using reverse transcription (the process of creating a single stranded RNA from a DNA molecule) together with amplification by polymerase chain reaction (PCR, a method used to rapidly make millions to billions of copies of a specific DNA sample), a process known as RT/PCR.
- This RT/PCR method was done on all virus segments at the same time, a technique known as multiplex RT/PCR.
- The primers for the RT/PCR were based on the conserved terminal sequences that are common in various influenza strains.
Usage of the Developed Method
- The RT/PCR products were found to be useful for determining partial nucleotide sequences. Nucleotides are organic molecules that are the basic structural units of nucleic acids such as DNA and RNA.
- It allows researchers to efficiently pinpoint the genetic origin of each segment by searching nucleotide sequence databases.
- The method enables scientists to quickly examine the genetic structure of viruses, opening up the potential for the development of treatments.
Application on H7N7 Equine Influenza Viruses
- In this study, the described method is specifically utilized in the investigation of genetic reassortment in H7N7 equine influenza viruses.
- Genetic reassortment in viruses refers to the process where viruses exchange or mix their genetic material.
- Understanding how gene reassortment happens in the H7N7 equine influenza virus could potentially help in predicting and controlling future outbreaks of this virus in horses.
Cite This Article
APA
Adeyefa CA, Quayle K, McCauley JW.
(1994).
A rapid method for the analysis of influenza virus genes: application to the reassortment of equine influenza virus genes.
Virus Res, 32(3), 391-399.
https://doi.org/10.1016/0168-1702(94)90087-6 Publication
Researcher Affiliations
- AFRC Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, UK.
MeSH Terms
- Base Sequence
- Conserved Sequence
- DNA Primers
- Databases, Factual
- Genes, Viral
- Influenza A virus / genetics
- Molecular Sequence Data
- Orthomyxoviridae / genetics
- Polymerase Chain Reaction / methods
- RNA, Viral / analysis
- RNA, Viral / genetics
- RNA-Directed DNA Polymerase
- Reassortant Viruses / genetics
- Sequence Alignment
- Sequence Analysis, DNA
- Sequence Homology, Nucleic Acid
Citations
This article has been cited 11 times.- Ladner JT, Sahl JW. Towards a post-pandemic future for global pathogen genome sequencing.. PLoS Biol 2023 Aug;21(8):e3002225.
- Van den Hoecke S, Verhelst J, Vuylsteke M, Saelens X. Analysis of the genetic diversity of influenza A viruses using next-generation DNA sequencing.. BMC Genomics 2015 Feb 14;16(1):79.
- Rash A, Woodward A, Bryant N, McCauley J, Elton D. An efficient genome sequencing method for equine influenza [H3N8] virus reveals a new polymorphism in the PA-X protein.. Virol J 2014 Sep 2;11:159.
- Murcia PR, Wood JL, Holmes EC. Genome-scale evolution and phylodynamics of equine H3N8 influenza A virus.. J Virol 2011 Jun;85(11):5312-22.
- Chen Q, Li J, Deng Z, Xiong W, Wang Q, Hu YQ. Comprehensive detection and identification of seven animal coronaviruses and human respiratory coronavirus 229E with a microarray hybridization assay.. Intervirology 2010;53(2):95-104.
- Zhou B, Donnelly ME, Scholes DT, St George K, Hatta M, Kawaoka Y, Wentworth DE. Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and Swine origin human influenza a viruses.. J Virol 2009 Oct;83(19):10309-13.
- Lodes MJ, Suciu D, Elliott M, Stover AG, Ross M, Caraballo M, Dix K, Crye J, Webby RJ, Lyon WJ, Danley DL, McShea A. Use of semiconductor-based oligonucleotide microarrays for influenza a virus subtype identification and sequencing.. J Clin Microbiol 2006 Apr;44(4):1209-18.
- Sengupta S, Onodera K, Lai A, Melcher U. Molecular detection and identification of influenza viruses by oligonucleotide microarray hybridization.. J Clin Microbiol 2003 Oct;41(10):4542-50.
- Cooper LA, Subbarao K. A simple restriction fragment length polymorphism-based strategy that can distinguish the internal genes of human H1N1, H3N2, and H5N1 influenza A viruses.. J Clin Microbiol 2000 Jul;38(7):2579-83.
- Adeyefa CA, McCauley JW, Tomori O. Mutational changes in the hemagglutinin of equine H3 influenza viruses result in the introduction of a glycosylation site which enhances the infectivity of the viruses.. Folia Microbiol (Praha) 1997;42(4):390-4.
- Adeyefa CA, James ML, McCauley JW. Antigenic and genetic analysis of equine influenza viruses from tropical Africa in 1991.. Epidemiol Infect 1996 Oct;117(2):367-74.
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