ABSTRACT
Infectious bronchitis (IB) is an acute, highly contagious contact respiratory disease of chickens caused by infectious bronchitis virus (IBV). IBV is very prone to mutation, which brings great difficulties to the prevention and control of the disease. Therefore, there is a pressing need for a method that is fast, sensitive, specific, and convenient for detecting IBV. In this study, a real-time fluorescence-based recombinase-aided amplification (RF-RAA) method was established. Primers and probe were designed based on the conserved regions of the IBV M gene and the reaction concentrations were optimized, then the specificity, sensitivity, and reproducibility of this assay were tested. The results showed that the RF-RAA method could be completed at 39â within 20â¯min, during which the results could be interpreted visually in real-time. The RF-RAA method had good specificity, no cross-reaction with common poultry pathogens, and it detected a minimum concentration of template of 2 copies/µL for IBV. Besides, its reproducibility was stable. A total of 144 clinical samples were tested by RF-RAA and real-time quantitative PCR (qPCR), 132 samples of which were positive and 12 samples were negative, and the coincidence rate of the two methods was 100â¯%. In conclusion, the developed RF-RAA detection method is rapid, specific, sensitive, reproducible, and convenient, which can be utilized for laboratory detection and clinical diagnosis of IBV.
Subject(s)
Chickens , Coronavirus Infections , Infectious bronchitis virus , Nucleic Acid Amplification Techniques , Poultry Diseases , Recombinases , Sensitivity and Specificity , Infectious bronchitis virus/genetics , Infectious bronchitis virus/isolation & purification , Animals , Chickens/virology , Poultry Diseases/virology , Poultry Diseases/diagnosis , Coronavirus Infections/diagnosis , Coronavirus Infections/veterinary , Coronavirus Infections/virology , Recombinases/metabolism , Recombinases/genetics , Reproducibility of Results , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/veterinary , DNA Primers/genetics , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/veterinary , Fluorescence , Molecular Diagnostic Techniques/methodsABSTRACT
Pest management has been a great challenge since the beginning of human agricultural activities. Since the 1930s, chemical pesticide control has been a major control technology that can solve some of the pest problems in agricultural production. Still, it is harmful to food safety and the ecological environment. Meanwhile, the extensive use of chemical pesticides may lead to the rapid development of pest resistance. Because of the advantages of low cost, eco-friendly advantage, and low side effects, Solar Insecticidal Lamp (SIL) as the main physical control technology has been widely used for pest management in agricultural production in China. Owing to the phototaxis of pests, they have a phototropic rhythm during the nighttime. We can adjust the SIL insecticidal time according to the phototropic rhythm of pests. The purpose of this paper is to provide a comprehensive review of the pest phototactic rhythm in a selection of 24 pest species. It is the first comprehensive survey on the phototactic rhythm of pests and the time segments of this survey are accurate to the hour. The phototactic rhythm of pests are investigated in two different varieties of crops: 1) food crops and 2) economic crops. We also discuss and analyze the various factors (e.g., meteorological conditions, insecticidal devices, physiological states and others) that affect the changing phototactic rhythm of pests. Finally, we highlight some open research challenge issues and future directions.
