Background: Bovine respiratory syncytial virus (BRSV) is a significant cause of bovine respiratory disease, resulting in significant losses to the cattle industry. For rapid detection of BRSV, a real-time recombinase-aided isothermal amplification assay (qRT-RAA) based on the F gene of BRSV was developed in this study. Results: The developed qRT-RAA assay showed good exponential amplification of the target fragment in 20 min at a constant temperature of 39 °C. And this assay displayed a high specificity for BRSV, without cross-reactions with Infectious Bovine Rhinotracheitis Virus (IBRV), Bovine Parainfluenza Virus Type 3 (BPIV3), Bovine Viral Diarrhea Virus (BVDV), and Bovine Coronavirus (BCoV). With the standard RNA of BRSV serving as a template, the limit of detection for qRT-RAA was 102 copies/μL. We examined ninety-seven clinical samples from cattle with respiratory disease using this method and determined a positive rate of 7.2% (7/97), consistent with results using the classical PCR method reported previously. Conclusions: A qRT-RAA assay for BRSV detection was established in this study. The method is specific and sensitive and can be completed within 20 min at 39 °C. These works demonstrate that the generated qRT-RAA assay is an effective diagnostic tool for rapidly detecting BRSV in resource-limited settings, which may be applied for the clinical detection of BRSV.

Elizabethkingia miricola is an emerging nosocomial pathogen responsible for meningitis, sepsis, urinary tract infection, pneumonia, and joint infection in humans. These pathogens were also reported to be causal agents for meningitis‐like disease in cultured frogs, which displayed high infectivity, mortality, and significant loss. In July 2023, 10 outbreaks of infectious meningitis‐like disease in bullfrogs occurred in Tangshan area. To determine the causal agent, 70 diseased frogs from 10 farms were collected for etiological identification. Gram‐negative bacilli were isolated from the brain and liver of sick bullfrogs and identified as members of E. miricola by biochemical characterization and 16S rRNA sequencing analysis. A total of 42 strains of E. miricola were isolated and further determined as the etiological agent by reproducing neurological symptoms and deaths in an artificial infection test. A representative isolate, HBTS‐1, was picked up for the pathogenicity test, and the data showed that this stain was highly pathogenic to bullfrogs with an LD₅₀ of 3.7 × 10⁵ CFU. Notably, the isolate also showed high pathogenicity to 5‐day‐old suckling mice, with an LD₅₀ of 3.1 × 10⁶ CFU, indicating its potential threat to mammals. Moreover, all the 42 E. miricola isolates showed resistance to multiple antibotics without an apparent inhibition zone observed in the test, making the choice of antimicrobial therapy challenging. These novel findings prioritized E. miricola as an important zoonotic agent, which may provide a reference for human medicine.