Avian Influenza Virus RNA Extraction from Tissue and Swab Material

2008 ◽  
pp. 13-18 ◽  
Author(s):  
Erica Spackman ◽  
David L. Suarez
2009 ◽  
Vol 54 (No. 9) ◽  
pp. 435-443 ◽  
Author(s):  
K. Rosenbergova ◽  
P. Lany ◽  
Z. Pospisil ◽  
O. Kubicek ◽  
V. Celer ◽  
...  

This study reports on the first quantification of avian influenza virus in the organs of mute swans that died during the epizootic of avian influenza (H5N1) between January and April 2006 in the Czech Republic. The quantitative real-time Reverse Transcriptase PCR (qRT-PCR) assay based on a TaqMan probe was developed for a rapid detection and quantification of avian influenza virus RNA in clinical samples collected from mute swans. Conserved regions in the matrix protein gene of avian influenza virus served as targets for the primers and TaqMan probe design. A recombinant plasmid containing the matrix protein gene amplicon was constructed for a quantitative assay of copy numbers of the target gene. Quantification of avian influenza virus RNA was accomplished using a standard curve generated from ten-fold serial dilutions of recombinant plasmid DNA in the range of 10<sup>2</sup> to 10<sup>8</sup> copies/µl. Avian influenza virus A/Cygnus olor/Brno-cz/2006 was adapted to grow in VERO cells. In the same passage of cell cultivation, the concentration of viral RNA was determined to be 1.01 × 10<sup>7</sup> copies/ml and TCID<sub>50</sub> was 10<sup>4.2</sup>/ml. From these values the ratio of one RNA copy to 0.00157 virion capable of VERO cells infection was calculated. This ratio was used to estimate the virus concentrations in the tissues of dead mute swans.


2008 ◽  
Vol 149 (1) ◽  
pp. 190-194 ◽  
Author(s):  
Julie L. Forster ◽  
Valerie B. Harkin ◽  
David A. Graham ◽  
Samuel J. McCullough

2002 ◽  
Vol 76 (15) ◽  
pp. 7860-7862 ◽  
Author(s):  
Thomas G. Fanning ◽  
Richard D. Slemons ◽  
Ann H. Reid ◽  
Thomas A. Janczewski ◽  
James Dean ◽  
...  

ABSTRACT Wild waterfowl captured between 1915 and 1919 were tested for influenza A virus RNA. One bird, captured in 1917, was infected with a virus of the same hemagglutinin (HA) subtype as that of the 1918 pandemic virus. The 1917 HA is more closely related to that of modern avian viruses than it is to that of the pandemic virus, suggesting (i) that there was little drift in avian sequences over the past 85 years and (ii) that the 1918 pandemic virus did not acquire its HA directly from a bird.


2015 ◽  
Vol 13 (1) ◽  
pp. 11-17 ◽  
Author(s):  
MZ Hassan ◽  
BC Das ◽  
MS Mahmud ◽  
MA Amin ◽  
MA Yousuf ◽  
...  

Waterfowl are the natural reservoir of avian influenza viruses and ducks may play a role in the maintenance of avian influenza type A. The aim of the present study was to investigate the seroprevalence and detection of avian influenza virus (AIV) type A in duck. This study was carried out during July 2013 to December 2013 on AIV type A from semi-scavenging farm at Nikli and Bajitpur upazila of Kishoregonj district in Bangladesh. A total of 368 blood samples were collected from duck and tested by indirect ELISA for seroprevalence. For detection of AIV type A, The cloacal swabs were collected from 75 duck and subjected to RNA extraction and real time RT-PCR (rRT-PCR) with specific primer and probe for detection of matrix (M) gene. The average seroprevalance of AIV type A in seven different age groups was found to be 90.21%. The highest (25.81 %) seroprevalence was found in 5 months age of birds and the lowest (2.44 %) was found in 12 months age of birds. As regard to area distribution, the average degree of seroprevalence was 93.51% from Nikli had the highest order than Bajitpur (86.88%) upazila of Bangladesh. In case of cloacal sample by using rRT–PCR, out of 15 pooling cloacal samples, two pooling samples (13.33%) that contain 10 samples were positive and 13 pooling samples showed negative (86.67%) for AIV type A in duck. It can be concluded that the long distance movement of duck flocks, may influence outbreak of avian influenza virus (AIV) type A among different poultry species in Bangladesh. Therefore, it needs to develop control strategy for future dissemination of AIV in duck population.DOI: http://dx.doi.org/10.3329/bjvm.v13i1.23705Bangl. J. Vet. Med. (2015). 13 (1): 5-9


2012 ◽  
Vol 56 (4s1) ◽  
pp. 955-958 ◽  
Author(s):  
Sasan R. Fereidouni ◽  
Anja Globig ◽  
Elke Starick ◽  
Timm C. Harder

2008 ◽  
Vol 82 (7) ◽  
pp. 3769-3774 ◽  
Author(s):  
Michael Worobey

ABSTRACT Zhang et al. (G. Zhang, D. Shoham, D. Gilichinsky, S. Davydov, J. D. Castello, and S. O. Rogers, J. Virol. 80:12229-12235, 2006) have claimed to have recovered influenza A virus RNA from Siberian lake ice, postulating that ice might represent an important abiotic reservoir for the persistence and reemergence of this medically important pathogen. A rigorous phylogenetic analysis of these influenza A virus hemagglutinin gene sequences, however, indicates that they originated from a laboratory reference strain derived from the earliest human influenza A virus isolate, WS/33. Contrary to Zhang et al.'s assertions that the Siberian “ice viruses” are most closely related either to avian influenza virus or to human influenza virus strains from Asia from the 1960s (Zhang et al., J. Virol. 81:2538 [erratum], 2007), they are clearly contaminants from the WS/33 positive control used in their laboratory. There is thus no credible evidence that environmental ice acts as a biologically relevant reservoir for influenza viruses. Several additional cases with findings that seem at odds with the biology of influenza virus, including modern-looking avian influenza virus RNA sequences from an archival goose specimen collected in 1917 (T. G. Fanning, R. D. Slemons, A. H. Reid, T. A. Janczewski, J. Dean, and J. K. Taubenberger, J. Virol. 76:7860-7862, 2002), can also be explained by laboratory contamination or other experimental errors. Many putative examples of evolutionary stasis in influenza A virus appear to be due to laboratory artifacts.


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