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A rapid cycleave PCR method for distinguishing the vaccine strain Brucella abortus A19 in China
- Source :
- Journal of Veterinary Diagnostic Investigation. 28:214-218
- Publication Year :
- 2016
- Publisher :
- SAGE Publications, 2016.
-
Abstract
- Brucellosis is a widespread zoonotic disease caused by Brucella spp. Immunization with attenuated vaccines has proved to be an effective method of prevention; however, it may also interfere with diagnosis. Brucella abortus strain A19, which is homologous to B. abortus strain S19, is widely used for the prevention of bovine brucellosis in China. For effective monitoring of the control of brucellosis, it is essential to distinguish A19 from field strains. Single-nucleotide polymorphism–based assays offer a new approach to such discrimination studies. In the current study, we developed a cycleave PCR assay that successfully distinguished attenuated vaccine strains A19 and S19 from 22 strains of B. abortus and 57 strains of 5 other Brucella species. The assay gave a negative reaction with 4 non- Brucella species. The minimum sensitivity of the assay, evaluated using 10-fold dilutions of chromosomal DNA, was 7.6 fg for the A19 strain and 220 fg for the single non-A19/non-S19 Brucella strain tested ( B. abortus 104M). The assay was also reproducible (intra- and interassay coefficients of variation: 0.003–0.01 and 0.004–0.025, respectively). The cycleave assay gave an A19/S19-specific reaction in 3 out of 125 field serum samples, with the same 3 samples being positive in an alternative A19/S19-specific molecular assay. The cycleave assay gave a total of 102 Brucella-specific reactions (3 being the A19/S19-specific reactions), whereas an alternative Brucella-specific assay gave 92 positive reactions (all also positive in the cycleave assay). Therefore, this assay represents a simple, rapid, sensitive, and specific tool for use in brucellosis control.
- Subjects :
- 0301 basic medicine
China
Serial dilution
Swine
030106 microbiology
Brucella Vaccine
Brucella abortus
Brucella
Biology
Polymerase Chain Reaction
complex mixtures
Brucellosis
law.invention
Microbiology
03 medical and health sciences
Predictive Value of Tests
law
medicine
Animals
Polymerase chain reaction
Attenuated vaccine
General Veterinary
Strain (chemistry)
Goats
Vaccination
medicine.disease
biology.organism_classification
Virology
030104 developmental biology
Cattle
Subjects
Details
- ISSN :
- 19434936 and 10406387
- Volume :
- 28
- Database :
- OpenAIRE
- Journal :
- Journal of Veterinary Diagnostic Investigation
- Accession number :
- edsair.doi.dedup.....cb2047a1d22e37dfc5f117b317425439