() is one of the primary agents involved in porcine respiratory disease complex, and circulates in the swine industry worldwide. The prevention and control of is complicated. Thus, a recombinase-aided amplification (RAA) assay coupled with the clustered regularly-interspaced short palindromic repeats (CRISPR)/Cas12a system was established for the detection of . The most suitable primer pairs and CRISPR RNA (crRNA) were screened and selected for the RAA-CRISPR/Cas12a detection system. We have achieved a detection limit of 1 copy/µL and 5 copies/µL per reaction for the RAA-CRISPR/Cas12a-fluorescence assay and RAA-CRISPR/Cas12a-lateral flow assay (LFA), respectively. Furthermore, the RAA-CRISPR/Cas12a system displayed no cross-reactivity with other respiratory pathogens. The performance of the RAA-CRISPR/Cas12a system was compared with PCR as recommended by the Chinese national standard (GB/T 35909-2018) and qPCR as recommended by the Chinese entry-exit inspection and quarantine industry standard (SN/T4104-2015) for clinical samples, and good consistency with these methods was observed. Above all, the methods shed a light on the convenient, portable, visual, highly sensitive and specific detection of , demonstrating a great application potential for on-site monitoring of in the field.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11695356 | PMC |
| http://dx.doi.org/10.3389/fcimb.2024.1469558 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Specific and sensitive detection of bovine coronavirus using CRISPR-Cas13a combined with RT-RAA technology.
Front Vet Sci
January 2025
Key Laboratory of Animal Medicine at Southwest Minzu University of Sichuan Province, College of Animal Science and Veterinary Medicine, Southwest Minzu University, Chengdu, China.
Introduction: Bovine coronavirus (BCoV) is an important pathogen of enteric and respiratory disease in cattle, resulting in huge economic losses to the beef and dairy industries worldwide. A specific and sensitive detection assay for BCoV is critical to the early-stage disease prevention and control.
Methods: We established a specific, sensitive, and stable assay for BCoV nucleic acid detection based on CRISPR/Cas13a combined with reverse transcription recombinase-aided amplification (RT-RAA) technology.
Establishment of a rapid method for the detection of canis based on recombinase-mediated thermostable nucleic acid amplification technology.
Front Cell Infect Microbiol
January 2025
The First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China.
Objective: To establish a rapid detection method for canine using recombinase-aided amplification (RAA) technology.
Methods: The outer membrane protein 25 gene fragment (Omp25) of canis was targeted. Primers and fluorescent probes were designed and synthesized, and recombinant plasmids were constructed as standards.
Rapid visual detection of hepatitis E virus combining reverse transcription recombinase-aided amplification with lateral flow dipstick and real-time fluorescence.
J Clin Microbiol
January 2025
Laboratory of Animal Pathology and Public Health, National Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Unlabelled: Hepatitis E virus (HEV) is a globally prevalent zoonotic pathogen that is primarily spread through the fecal-oral route, such as by consuming undercooked or contaminated pork. HEV infection leads to an estimated 3.3 million symptomatic cases of viral hepatitis and 70,000 deaths in human populations each year.
View Article and Find Full Text PDFUsing Recombinase-Aid Amplification Combined with Argonaute for Rapid Sex Identification in Flamingo ().
Animals (Basel)
December 2024
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
Flamingos () are among the oldest birds worldwide and are loved by people for their bright red feathers. In addition, flamingos are sexually monomorphic birds, and distinguishing between males and females is difficult. The polymerase chain reaction (PCR) is widely used for sex identification.
View Article and Find Full Text PDFRapid Detection of Feline Calicivirus Using Lateral Flow Dipsticks Based on CRISPR/Cas13a System.
Animals (Basel)
December 2024
The College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China.
Feline calicivirus (FCV) is one of the most common viral pathogens in domestic cats worldwide, which mainly causes upper respiratory tract infections in felines and seriously threatens the health of felines. Consequently, it is crucial to establish a rapid detection method to efficiently take control and prevent the spread of FCV. To construct the Cas13a-RAA-LFD reaction system, this study specifically designed recombinase-aided amplification (RAA) primers added with a T7 promoter and CRISPR RNA (crRNA), which were both based on the FCV relatively conserved sequence.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!