Multiplex nested RT-PCR for detecting avian influenza virus, infectious bronchitis virus and Newcastle disease virus

2013 ◽  
Vol 188 (1-2) ◽  
pp. 41-46 ◽  
Author(s):  
Thanh Trung Nguyen ◽  
Hyuk-Joon Kwon ◽  
Il-Hwan Kim ◽  
Seung-Min Hong ◽  
Won-Jin Seong ◽  
...  
2020 ◽  
Vol 11 (1) ◽  
pp. 1-6
Author(s):  
Amjad Ali Channa ◽  
Nazeer Hussain Kalhoro ◽  
Zaheer Ahmed Nizamani ◽  
Ayaz Hussain Mangi ◽  
Jamila Soomro

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.


2019 ◽  
Vol 1 (2) ◽  
Author(s):  
Feng Fei

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.


2016 ◽  
Vol 60 (1s) ◽  
pp. 191-201 ◽  
Author(s):  
Amélyne Lardinois ◽  
Olivier Vandersleyen ◽  
Mieke Steensels ◽  
Nathalie Desloges ◽  
Jan Mast ◽  
...  

2013 ◽  
Vol 8 (6) ◽  
pp. 520-526 ◽  
Author(s):  
Dawid Nidzworski ◽  
Krzysztof Smietanka ◽  
Zenon Minta ◽  
Bogusław Szewczyk

AbstractNewcastle disease Virus (NDV), a member of the Paramyxoviridae family, and Influenza virus, from the Orthomyxoviridae family, are two main avian pathogens that cause serious economic problems in poultry farming. NDV strains are classified into three major pathotypes: velogenic, mesogenic, and lentogenic. Avian influenza viruses (AIV) are also divided into: low pathogenic (LPAI) and highly pathogenic (HPAI) strains. Both viruses are enveloped, single stranded, negative-sense RNA viruses which give similar symptoms ranging from sub-clinical infections to severe disease, including loss in egg production, acute respiratory syndrome, and high mortality, depending on their level of pathogenicity. This similarity hinders diagnosis when based solely on clinical and post mortem examination. Most of the currently available molecular detection methods are also pathogenspecific, so that more than one RT-PCR is then required to confirm or exclude the presence of both pathogens. To overcome this disadvantage, we have applied a One Step Duplex RT-PCR method to distinguish between those two pathogens. The main objective of the project was to develop a universal, fast, and inexpensive method which could be used in any veterinary laboratory.


Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1606
Author(s):  
Xiaorong Zhang ◽  
Zongyi Bo ◽  
Chenchen Meng ◽  
Yin Chen ◽  
Chengcheng Zhang ◽  
...  

H9N2 avian influenza virus (AIV) has become endemic in many countries, causing great economic losses when co-infected with other pathogens. So far, several live vaccines based on Newcastle disease virus (NDV) vectors expressing influenza hemagglutinin (HA) have been developed. However, the thermostable recombinant NDV is rarely reported. In this study, using a thermostable NDV rAHR09 strain as the vector, three recombinant NDVs expressing native HA, chimeric HA ectodomain with transmembrane domain/C-terminal cytoplasmic tail domain from fusion protein of NDV, and HA ectodomain were generated, designated rAHR09-HA, rAHR09-HAF, and rAHR09-HAE. The MDT value of three recombinant NDVs was above 120 h, their ICPI value was about 0.03, and the recombinant NDVs were still infectious when treated for 100 min under 56 °C, which demonstrated that the recombinant NDVs kept the lentogenic and thermostable nature of rAHR09. The immunization data showed that rAHR09-HA and rAHR09-HAF induced a higher HI antibody titer against H9N2 AIV and NDV. After being challenged with H9N2 AIV, the rAHR09-HA and rAHR09-HAF could significantly reduce the virus shedding in cloacal and tracheal swab samples. Our results suggest that rAHR09-HA and rAHR09-HAF might be vaccine candidates against H9N2 AIV.


Sign in / Sign up

Export Citation Format

Share Document