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A novel approach for detection of brucella using a real-time recombinase polymerase amplification assay
- Source :
- Molecular and cellular probes. 48
- Publication Year :
- 2019
-
Abstract
- Brucella, the etiological agent of brucellosis, is an important zoonosis pathogen worldwide. Brucella infects humans and various domestic and wild animals, and represents a great threat to public health and animal husbandry. In the present study, we developed a real-time recombinase polymerase amplification (RPA) assay for the detection of Brucella. The assay targeted the bcsp31 gene of Brucella, and an RPA exo probe and a pair of primers were selected for assay validation. RPA sensitivity and specificity were evaluated using plasmid standards, Brucella representative strains, and non-Brucella strains. The RPA assay achieved a detection limit of 17 molecules in 95% of cases based on probit analysis, and could successfully distinguish 18 representative Brucella strains (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae and B. ovis), and four Brucella vaccine strains (A19, S19, S2 and M5). A total of 52 Brucella field strains were detected by real-time PCR and RPA in parallel, and compared with real-time PCR, the sensitivity of the RPA assay was 94% (49/52). Thus, this RPA assay may be a rapid, sensitive, and specific tool for the prevention and control of Brucellosis.
- Subjects :
- DNA, Bacterial
Recombinase Polymerase Amplification
Brucella
Real-Time Polymerase Chain Reaction
complex mixtures
Sensitivity and Specificity
Recombinases
03 medical and health sciences
Plasmid
Bacterial Proteins
medicine
Molecular Biology
Gene
Pathogen
030304 developmental biology
DNA Primers
0303 health sciences
Brucella Vaccine
biology
030306 microbiology
Zoonosis
Brucellosis
Cell Biology
bacterial infections and mycoses
medicine.disease
biology.organism_classification
Virology
enzymes and coenzymes (carbohydrates)
Subjects
Details
- ISSN :
- 10961194
- Volume :
- 48
- Database :
- OpenAIRE
- Journal :
- Molecular and cellular probes
- Accession number :
- edsair.doi.dedup.....c76291f5f3a11c9792b79b422db11386