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.

Real-time RT-PCR assay to differentiate clades of H5N1 avian influenza viruses circulating in Vietnam

2013 ◽  
Vol 193 (2) ◽  
pp. 452-458 ◽  
Author(s):  
Z. Kis ◽  
J. Jones ◽  
A. Creanga ◽  
K. Ferdinand ◽  
K. Inui ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay ◽  
H5n1 Avian Influenza

Development of a TaqMan MGB RT-PCR assay for the detection of type A and subtype H10 avian influenza viruses

2018 ◽  
Vol 163 (9) ◽  
pp. 2497-2501 ◽  
Author(s):  
Fan Yang ◽  
Bin Chen ◽  
Fumin Liu ◽  
Xiuming Peng ◽  
Tao Sun ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Type A ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay

Multiple RT-PCR Detection of H5, H7, and H9 Subtype Avian Influenza Viruses and Newcastle Disease Virus

2019 ◽  
Vol 1 (2) ◽  
Author(s):  
Feng Fei
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Newcastle Disease Virus ◽  
Avian Influenza Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Pcr Detection ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses

Objective: This paper focuses on the multiple detection RT-PCR technology of H5, H7, AND H9 subtype avian influenza viruses and Newcastle disease virus, and points out the specific detection methods and detection procedures of avian influenza and Newcastle disease virus. Methods: The genes of Newcastle disease virus carrying out the HA gene sequence of H5, H7 and H9 subtype AIV in GenBank were used to establish a strategy for simultaneous detection of three subtypes of avian influenza virus and Newcastle disease virus. Results: The results showed that the program can detect and distinguish H5, H7 and H9 subtype avian influenza viruses and Newcastle disease virus at one time. Conclusion: Multiple RT-PCR detection method has high detection sensitivity and can detect and determine different subtypes of avian influenza virus and Newcastle disease virus quickly and accurately, therefore, it has a crucial role in the detection and control of avian influenza H5, H7 and H9 subtypes and Newcastle disease.

Development of a real-time TaqMan RT-PCR assay for the detection of H9N2 avian influenza viruses

2010 ◽  
Vol 168 (1-2) ◽  
pp. 72-77 ◽  
Author(s):  
M. Ben Shabat ◽  
R. Meir ◽  
R. Haddas ◽  
E. Lapin ◽  
I. Shkoda ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay

scholarly journals A duplex RT-PCR assay for detection of H9 subtype avian influenza viruses and infectious bronchitis viruses

2016 ◽  
Vol 15 (9) ◽  
pp. 2105-2113 ◽  
Author(s):  
Yan-di WEI ◽  
Wei-hua GAO ◽  
Hong-lei SUN ◽  
Chen-fang YU ◽  
Xing-yao PEI ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay ◽  
Infectious Bronchitis

Application of real-time rt-pcr utilising lux (light upon extension) fluorogenic primer for the rapid detection of avian influenza viruses

2006 ◽  
Vol 54 (4) ◽  
pp. 525-533 ◽  
Author(s):  
I. Kiss ◽  
P. Germán ◽  
L. Sámi ◽  
Márta Antal ◽  
T. Farkas ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Allantoic Fluid ◽  
Cost Effective ◽  
Influenza Viruses ◽  
Influenza Surveillance ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay ◽  
Efficient Detection

A real-time RT-PCR assay utilising light upon extension fluorogenic primer (LUX RT-PCR) was developed for the rapid and efficient detection of avian influenza viruses (AIV). The assay detected each of the AIV isolates tested (16/16) and gave negative results with heterologous pathogens (17/17). The detection limit of the assay proved to be 10-0.5 EID50/0.2 ml and 101.5 EID50/0.2 ml in allantoic fluid of virus-infected embryonated chicken eggs and in spiked chicken faeces samples, respectively. Based on its specificity, sensitivity and relative simplicity, the LUX RT-PCR assay provides a novel, rapid and cost-effective diagnostic tool for avian influenza surveillance and monitoring programs.

Establishment of a multiplex real-time RT-PCR assay for rapid identification of H6 subtype avian influenza viruses

2018 ◽  
Vol 163 (6) ◽  
pp. 1671-1675 ◽  
Author(s):  
Fan Yang ◽  
Haibo Wu ◽  
Fumin Liu ◽  
Xiangyun Lu ◽  
Xiuming Peng ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Rapid Identification ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay

scholarly journals Genetic Characterization and Pathogenicity of H7N7 and H7N9 Avian Influenza Viruses Isolated from South Korea

Viruses ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2057
Author(s):  
Eun-Jee Na ◽  
Young-Sik Kim ◽  
Yoon-Ji Kim ◽  
Jun-Soo Park ◽  
Jae-Ku Oem
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
South Korea ◽  
Influenza Viruses ◽  
The Body ◽  
Avian Influenza Viruses ◽  
Receptor Binding Site ◽  
Phylogenetic Tree Analysis ◽  
Pathogenic Avian Influenza ◽  
Ha Gene

H7 low pathogenic avian influenza viruses (LPAIVs) can mutate into highly pathogenic avian influenza viruses (HPAIVs). In addition to avian species, H7 avian influenza viruses (AIVs) also infect humans. In this study, two AIVs, H7N9 (20X-20) and H7N7 (34X-2), isolated from the feces of wild birds in South Korea in 2021, were genetically analyzed. The HA cleavage site of the two H7 Korean viruses was confirmed to be ELPKGR/GLF, indicating they are LPAIVs. There were no amino acid substitutions at the receptor-binding site of the HA gene of two H7 Korean viruses compared to that of A/Anhui/1/2013 (H7N9), which prefer human receptors. In the phylogenetic tree analysis, the HA gene of the two H7 Korean viruses shared the highest nucleotide similarity with the Korean H7 subtype AIVs. In addition, the HA gene of the two H7 Korean viruses showed high nucleotide similarity to that of the A/Jiangsu/1/2018(H7N4) virus, which is a human influenza virus originating from avian influenza virus. Most internal genes (PB2, PB1, PA, NP, NA, M, and NS) of the two H7 Korean viruses belonged to the Eurasian lineage, except for the M gene of 34X-2. This result suggests that active reassortment occurred among AIVs. In pathogenicity studies of mice, the two H7 Korean viruses replicated in the lungs of mice. In addition, the body weight of mice infected with 34X-2 decreased 7 days post-infection (dpi) and inflammation was observed in the peribronchiolar and perivascular regions of the lungs of mice. These results suggest that mammals can be infected with the two H7 Korean AIVs. Our data showed that even low pathogenic H7 AIVs may infect mammals, including humans, as confirmed by the A/Jiangsu/1/2018(H7N4) virus. Therefore, continuous monitoring and pathogenicity assessment of AIVs, even of LPAIVs, are required.

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