Development of a Novel Loop-Mediated Isothermal Amplification (LAMP) for Rapid Diagnosis of Newcastle Disease in Field and Resource-Limited Areas

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Background: Newcastle Disease (ND) is a highly contagious and fatal poultry disease caused by Newcastle disease virus (NDV) and has a global distribution. To control this disease, a rapid method for diagnosing it is needed. PCR-based molecular diagnostics such as real-time reverse transcription-PCR (RRT-PCR) for detecting NDV genes have been conducted worldwide. However, because PCR methods are time-consuming and require a well-equipped laboratory, they are not well-suited for use in field and resource limited areas. Methods: In this study, we established a field-friendly loop-mediated isothermal amplification (LAMP) assay for NDV diagnosis using primers targeting the highly conserved L gene, which bypasses the need for RNA extraction and makes it more suitable for field application than previously reported LAMP methods. Results: Our LAMP method is capable of detecting a broad range of NDV genotypes and showed no cross-reactivity with other avian viral diseases or the host genome. The reaction is completed within 35 minutes of incubation at 65°C. NDV was successfully detected directly from swab and tissue samples without the need for conventional RNA extraction. Conclusion: The LAMP method developed in this study offers a rapid, affordable, and field-friendly diagnostic tool for NDV detection.

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