scholarly journals A modified strategy of multiplex RT-PCR for simultaneous detection of H5, H7, and H9 subtypes of avian influenza virus based on common forward oligo

2016 ◽  
Vol 25 (3) ◽  
pp. 322-327 ◽  
Author(s):  
M.S. Tahir ◽  
D. Mehmood ◽  
A.U. Sultan ◽  
M.H. Saeed ◽  
A.R. Khan ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Simultaneous Detection ◽  
Rt Pcr

Rapid identification of H5 avian influenza virus in chicken throat swab specimens using microfluidic real-time RT-PCR

2014 ◽  
Vol 6 (8) ◽  
pp. 2628 ◽  
Author(s):  
Ling Zhu ◽  
Cancan Zhu ◽  
Guoqing Deng ◽  
Long Zhang ◽  
Shumi Zhao ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Real Time ◽  
Throat Swab ◽  
Rapid Identification ◽  
Rt Pcr

scholarly journals Development of a duplex TaqMan real-time RT-PCR assay for simultaneous detection of newly emerged H5N6 influenza viruses

2019 ◽  
Vol 16 (1) ◽  
Author(s):  
Lin Liu ◽  
Ying Zhang ◽  
Pengfei Cui ◽  
Congcong Wang ◽  
Xianying Zeng ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Simultaneous Detection ◽  
Reaction System ◽  
Taxonomic Status ◽  
Human Infection ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay

Abstract Background In 2017–2018, a new highly pathogenic H5N6 avian influenza virus (AIV) variant appeared in poultry and wild birds in Asian and European countries and caused multiple outbreaks. These variant strains are different from the H5N6 virus associated with human infection in previous years, and their genetic taxonomic status and antigenicity have changed. Therefore, revision of the primers and probes of fluorescent RT-PCR is important to detect the new H5N6 subtype AIV in poultry and reduce the risk of an epidemic in birds or humans. Methods In this study, the primers and probes including three groups of HA and four groups of NA for H5N6 influenza virus were evaluated. Then a set of ideal primer and probes were selected to further optimize the reaction system and established a method of double rRT-PCR assay. The specificity of this method was determined by using H1~H16 subtype AIV. Results The results showed that fluorescence signals were obtained for H5 virus in FAM channel and N6 virus in VIC channel, and no fluorescent signal was observed in other subtypes of avian influenza viruses. The detection limit of this assay was 69 copies for H5 and 83 copies for N6 gene. And, the variability tests of intra- and inter-assay showed excellent reproducibility. Moreover, this assay showed 100% agreement with virus isolation method in detecting samples from poultry. Conclusion The duplex rRT-PCR assay presented here has high specificity, sensitivity and reproducibility, and can be used for laboratory surveillance and rapid diagnosis of newly emerged H5N6 subtype avian influenza viruses.

Development of a TaqMan MGB RT-PCR for the rapid detection of H3 subtype avian influenza virus circulating in China

2015 ◽  
Vol 217 ◽  
pp. 64-69 ◽  
Author(s):  
Qiaoyang Teng ◽  
Weixia Shen ◽  
Dawei Yan ◽  
Liping Yan ◽  
Xuesong Li ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Rapid Detection ◽  
Rt Pcr ◽  
H3 Subtype

A FULLY AUTOMATED PROCEDURE FOR THE HIGH THROUGHPUT DETECTION OF AVIAN INFLUENZA VIRUS BY REAL-TIME RT-PCR

2007 ◽  
Vol 2 (s1) ◽  
pp. e13-e13
Author(s):  
Montserrat Agüero ◽  
Elena San Miguel ◽  
Azucena Sánchez ◽  
Concepción Gómez-Tejedor ◽  
Miguel Angel Jiménez-Clavero
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Real Time ◽  
High Throughput ◽  
Rt Pcr ◽  
High Throughput Detection

scholarly journals Prevalence of Newcastle Disease Virus and Avian Influenza Virus (H7N3) in Poultry at Karachi

2020 ◽  
Vol 11 (1) ◽  
pp. 1-6
Author(s):  
Amjad Ali Channa ◽  
Nazeer Hussain Kalhoro ◽  
Zaheer Ahmed Nizamani ◽  
Ayaz Hussain Mangi ◽  
Jamila Soomro
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Newcastle Disease Virus ◽  
Avian Influenza Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Virus Isolation ◽  
Economic Losses ◽  
Rt Pcr ◽  
Agar Gel

Background: Poultry is largest and rapidly growing sector of livestock in Pakistan. It is mainly influenced by viral pathogens such as Newcastle Disease Virus (NDV) and Avian Influenza Virus (H7N3). These viruses cause severe disease in poultry and leads to heavy economic losses throughout the world. The outbreaks of these pathogens have been increased in last few decades. Therefore, the study about antigenic prevalence is needed to know about the emergence of these pathogenic viruses, and to get rid of severe ailments associated with reduced poultry production. Objectives: To determine the prevalence of Newcastle Disease Virus (NDV), Avian Influenza Virus (H7N3) and co-infections in poultry flocks at Karachi. Methodology: For detection of NDV and H7N3, a total of 200 tracheal swabs were collected and tested through virus isolation (V.I); the sample with positive virus isolation were tested through agar gel precipitation (AGP) and then the RNA was isolated through TRI Reagent, which was further tested through reverse transcription polymerase chain reaction (RT-PCR). Results: The virus isolation showed that 58% of samples were positive for various viruses. Agar gel precipitation (AGP) revealed that the occurrence of NDV, H7N3 and ND+H7 were 50%, 8% and 38%, respectively. RT-PCR for F and HA gene of NDV and H7N3 confirmed the presence of NDV and H7N3 in the poultry. Conclusion: It is concluded that NDV and H7N3 are circulating in the flocks causing co-infections, therefore it is important to know the field challenge of viruses and to prepare vaccine of circulating serotype of virus to mitigate the rate of infection.

scholarly journals Development of multiplex reverse transcription polymerase chain reaction (RT-PCR) for simultaneous detection of matrix, haemagglutinin and neuraminidase genes of H5N1 avian influenza virus

2012 ◽  
Vol 28 (2) ◽  
pp. 55-59 ◽  
Author(s):  
R. Mojumder ◽  
E. H. Chowdhury ◽  
R Parvin ◽  
J. A. Begum ◽  
M. Giasuddin ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Influenza Virus ◽  
Avian Influenza ◽  
Matrix Protein ◽  
Simultaneous Detection ◽  
High Fidelity ◽  
Rt Pcr ◽  
Chain Reaction ◽  
One Step ◽  
Polymerase Chain

Influenza A virus, subtype H5N1 causes a fatal disease in domestic poultry and could spread directly from poultry to humans. The aim of this study was to develop a multiplex reverse transcription polymerase chain reaction (mRT-PCR) for simultaneous detection of Type A influenza virus-specific matrix protein (M) gene as well as H5 and N1 genes of highly pathogenic avian influenza (HPAI) viruses. Finnzymes Phusion-Flash High- Fidelity PCR Master Mix (Finnzymes Oy, Finland) and Qiagen one-step RT-PCR enzyme mix (Qiagen, Germany) were used in a one-step RT-PCR. RNA was extracted from two known positive samples using Qiagen RNA extraction kit. RT-PCR was carried out with a mixture of primers specific for the Type A influenza virus matrix protein (M), and H5 and N1 genes of H5N1 HPAI viruses in a single reaction system. The mRT-PCR cDNA products were visualized by gel electrophoresis. The mRT-PCR yielded fragments of 245 bp for M, 545 bp for H5 and 343 bp for N1 genes of HPAI virus, which were clearly distinguishable. The mRT-PCR using the Finnzymes Phusion-Flash High-Fidelity PCR Master Mix (Finnzymes Oy, Finland) with Qiagen one-step RT-PCR Enzyme Mix (Qiagen, Germany) required only one hour and 20 minutes. (Bangl. vet. 2011. Vol. 28, No. 2, 55 – 59)DOI: http://dx.doi.org/10.3329/bvet.v28i2.10653

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