Reverse transcriptase-polymerase chain reaction-enzyme linked immunosorbent assay for rapid detection of avian influenza virus subtype H9N2

2006 ◽  
Vol 151 (12) ◽  
pp. 2447-2459 ◽  
Author(s):  
B. Chaharaein ◽  
A. R. Omar ◽  
I. Aini ◽  
K. Yusoff ◽  
S. S. Hassan
2017 ◽  
Vol 48 (3) ◽  
pp. 882-885 ◽  
Author(s):  
Trine Hammer Jensen ◽  
Jannie Holmegaard Andersen ◽  
Charlotte Kristiane Hjulsager ◽  
Mariann Chriél ◽  
Mads Frost Bertelsen

2007 ◽  
Vol 51 (s1) ◽  
pp. 235-241 ◽  
Author(s):  
Montserrat Agüero ◽  
Elena San Miguel ◽  
Azucena Sánchez ◽  
Concepción Gómez-Tejedor ◽  
Miguel Angel Jiménez-Clavero

2016 ◽  
Vol 43 (02) ◽  
pp. 75-79
Author(s):  
Lih-Chiann Wang ◽  
Wei-En Hsu ◽  
Wei Thong ◽  
Ting-Yen Chao ◽  
Ching-Ho Wang

Reverse transcription polymerase chain reaction (RT-PCR) was used routinely to detect the avian influenza virus (AIV) nucleoprotein (NP) gene. The purpose of the present study was to compare the correctness of a nested RT-PCR (nRT-PCR), one conventional RT-PCR with its outer primer (oRT-PCR) and the other conventional RT-PCR with its inner primer (iRT-PCR) to detect AIV NP gene. A total of 365 AI-free fecal swabs (73 pools), 7 tracheal swabs and anllantoic fluid from 25 chicken embryos were used to determine the analytic specificities of those tests. Compared with the iRT-PCR, the nRT-PCR was more sensitive for AIV detection. However, the specificities of nRT-PCR, oRT-PCR and iRT-PCR were 48.6% (35/72), 100% (67/67) and 91.3% (84/92), respectively. The amplifying band was sequenced and confirmed to be the AIV NP gene as the positive control. The specificity of this nRT-PCR is too low to be used for the AIV screening test.


Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1010
Author(s):  
Samah Mosad Mosad ◽  
Fatma A. El-Gohary ◽  
Hanaa Said Ali ◽  
Hanem El-Sharkawy ◽  
Ehab Kotb Elmahallawy

Avian influenza virus (AIV) remains one of the enzootic zoonotic diseases that challenges the poultry industry in Egypt. In the present study, a total of 500 tissue samples were collected from 100 chicken farms (broilers and layers) suspected to be infected with AIV through the period from 2009 to 2019 from Dakahlia governorate, Egypt. These samples were pooled in 100 working samples and screened for AIV then the positive samples were subjected to histopathological examination combined with real time-polymerase chain reaction (RRT-PCR). RRT-PCR positive samples were also subjected to conventional reverse transcriptase-polymerase chain reaction (RT-PCR) for detection of H5 AIV and some of these resulting positive samples were sequenced for detection of the molecular nature of the studied virus. Interestingly, the histopathological examination revealed necrotic liver with leukocytic infiltration with degenerative changes with necrotic pancreatitis, edema, and intense lymphoid depletion of splenic tissue and hyperplastic tracheal epithelium. Likewise, edema and congested sub mucosal blood vessels and intense bronchial necrosis with hyalinized wall vascular wall and heterophils infiltration were reported. Pneumonic areas with intense leukocytic aggregation mainly and vasculitis of the pulmonary blood vessels were also detected in lung. Collectively, these significant pathological changes in examined tissues cohered with AIV infection. Regarding the molecular characterization, 66 samples were positive for AIV by RRT-PCR and 52 of them were positive for H5 AIV by RT-PCR. The phylogenetic analysis revealed that the H5 viruses identified in this study were aligned with other Egyptian H5N1 AIVs in the Egyptian sub clade 2.2.1, while some of the identified strains were aligned with other Egyptian H5N8 strains in the new Egyptian sub clade 2.3.4.4. Taken together, our present findings emphasize the wide spread of AIV in Egypt and the importance of developing an efficient surveillance and periodical screening program for controlling such disease of public health concern.


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