Application of recombinase polymerase amplification method for rapid detection of infectious laryngotracheitis virus

Mol Cell Probes. 2020 Dec:54:101646. doi: 10.1016/j.mcp.2020.101646. Epub 2020 Aug 3.

Abstract

Infectious laryngotracheitis is a significant respiratory disease of chickens that causes huge economic losses due to high morbidity and mortality and reduced egg production. A real-time recombinase polymerase amplification (RPA) assay was developed to accurately detect ILTV. The specific probe and primer sets were carefully designed and screened. The real-time RPA assay was carried out at 39 °C for 30 min, and results were obtained within 15 min. The results of the specificity assay showed no fluorescence signals with other avian-related viruses. The sensitivity of the assay was 1 × 102 copies/μL. The low CV value showed that the assay was reproducible. A total of 115 clinical samples were tested using the real-time RPA assay and the real-time PCR assay in parallel; the coincidence rates of the two detection methods were 100%. The results indicated that the real-time RPA assay is a specific, sensitive, rapid, and useful tool for epidemiological studies and clinical diagnosis, especially in the field and in resource-poor areas.

Keywords: Detection; Infectious laryngotracheitis virus; Real-time; Recombinase polymerase amplification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers / metabolism
  • Herpesvirus 1, Gallid / genetics*
  • Herpesvirus 1, Gallid / isolation & purification*
  • Linear Models
  • Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction
  • Recombinases / metabolism*
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Time Factors

Substances

  • DNA Primers
  • Recombinases