Recombinase polymerase amplification–nucleic acid lateral flow immunoassays for Newcastle disease virus and infectious bronchitis virus detection

Abstract

Newcastle disease virus (NDV) and infectious bronchitis virus (IBV) are two poultry pathogens affecting the respiratory tract of chickens, and cause major economic losses in the industry. Rapid detection of these viruses is crucial to inform implementation of appropriate control measures. The objective of our study is developing a simple, rapid and field applicable recombinase polymerase amplification (RPA)-nucleic acid lateral flow (NALF) immunoassay for detection of NDV and IBV. Isothermal amplification of the matrix protein (M) gene of NDV and the nucleoprotein (N) gene of IBV was implemented via recombinase polymerase amplification at 38 °C for 40 min and 20 min, respectively using modified labeled primers. NALF device used in this study utilizes antibodies for detection of labeled RPA amplicons. The results revealed that RPA-NALF immunoassays can detect both viruses after 40 min at 38 °C and only NDV after 20 min. The limit of detection (LOD) was 10 genomic copies/RPA reaction. The assays results on clinical samples collected from diseased chicken farms demonstrated a good performance in comparison with quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). The assays established in this study can facilitate rapid, on-site molecular diagnosis of suspected cases of ND and IB viral infections as the results can be detected by the naked eye without the need for measuring fluorescence. Furthermore, the NALF device could be adapted to detect other infectious agents.

A Recombinant La Sota Vaccine Strain Expressing Multiple Epitopes of Infectious Bronchitis Virus (IBV) Protects Specific Pathogen-Free (SPF) Chickens against IBV and NDV Challenges

“…IBV-inactivated vaccines also face some defects such as lack of ability to elicit strong cellular immune responses and lack of cross-immunoprotective effects. Furthermore, chicken flocks suffer co-infection with IBV and avian orthoavulavirus-1 (AOAV-1), which is a member of the genus Orthoavulavirus of the family Paramyxoviridae, often referred to as Newcastle disease virus (NDV) [15], which hinders the prevention and control of these diseases [16,17]. These two pathogens remain a vital threat to domestic poultry production [18][19][20][21].…”

Early Stage Development of a Newcastle Disease Vaccine Candidate in Corn

“…Newcastle disease also has devastating effects on domestic poultry production ( 2 ). The spread of diseases in wild and domestic birds can only be prevented through timely disease diagnosis and mass application of effective vaccines ( 1 , 3 ). Newly developed plant-based vaccines are efficient, affordable, easy to produce, convenient to store and transport, and appropriate for immunizing a large number of farm animals through feed; thus, plant-based vaccines can prevent economic losses caused by the outbreak of this notorious disease ( 4 – 7 ).…”

Development of a Recombinant Thermostable Newcastle Disease Virus (NDV) Vaccine Express Infectious Bronchitis Virus (IBV) Multiple Epitopes for Protecting against IBV and NDV Challenges

“…Avian Newcastle disease (ND) and infectious bronchitis (IB) caused by the Newcastle disease virus (NDV) and infectious bronchitis virus (IBV) are able to widely spread in chickens and give rise to massive losses in the poultry industry around the world. [ 1 , 2 , 3 ]. NDV and IBV belong to the family Paramyxoviridae and Coronaviridae, respectively, and the whole genome is approximately 15.2 kb and 27.6 kb in size.…”