Single-tube duplex-PCR for specific detection and differentiation of Brucella abortus S19 vaccine strain from other Brucella spp

Abstract

A simple single-tube duplex-PCR assay was optimized for rapid and sensitive detection and differentiation of Brucella abortus (B.abortus) S19 vaccine strain from other Brucella spp.(B.abortus 544,B abortus S99,B.melitensis 16M,B.suis).This assay was optimized using two primer pairs that were designed, one targeting genus specific multicopy IS711 and another eryC for the development of the duplex-PCR assay. Specificity of the assay was assessed using DNA templates from various Brucella species (B.abortus, B.melitensis, B.suis) and non Brucella bacteria like Staphylococcus aureus, Proteus vulgaris, Pasteurella multocida, Pseudomonas aeruginosa, Citrobacter freundii, E. coli, Salmonella enteritidis, Campylobacter jejuni and Listeria monocytogenes. The assay was also evaluated on spiked milk samples with known B.abortus cultures. The assay was able to detect up to 2.47 x 103CFU of B. abortus S99 organism in spiked milk sample. The assay was further validated in 53 clinical specimens (aborted foetal stomach content). Results confirm that Single tube duplex-PCR assay was useful for early and rapid detection and differentiation of B. abortus vaccine strain S19 from other Brucella spp in milk as well as in clinical specimen.

aspase-11 plays an important role in IL-1, IL-18 and IL-1â secretion from porcine alveolar macrophage cells stimulated with Brucella suis LPS

“…In the early stages of Brucella infection, it minimizes the host pro-inflammatory response, and attempt to establish the mechanism of chronic infection, which allows the Brucella intracellular survive and replication (Aparajita et al, 2018, Roset et al, 2013. The research identifying the key cellular regulators of cytokine secretion of macrophages under the stimulation of Brucella LPS will helpful to the development of novel anti-inflammatory targets and specific anti-inflammatory adjunctive therapies of brucellosis.…”

Milk Borne Brucellosis

“…Over the past few decades, small ruminant and bovine brucellosis control programs have had resounding success because of the use of two live vaccines: B. melitensis Rev. 1 and B. abortus S19 [60]. B. abortus RB51 is suggested as a vaccine for bovine brucellosis to be applied in the last stages of protective programs in addition to tests and slaughter [61].…”

Gated nanoprobe utilizing metal–organic frameworks for identifying and distinguishing between the wild strains and the vaccine strains of brucella

“…Unfortunately, due to individual differences, the false negative rate is high in these methods. 8 Quantitative Real-time PCR (qPCR) 9 is a widely utilized method for quantifying gene expression. It has been employed to distinguish between wild strains of Brucella and B. suis S19 vaccine strains.…”

“…Quantitative Real-time PCR (qPCR) 9 is a widely utilized method for quantifying gene expression. It has been employed to distinguish between wild strains of Brucella and B. suis S19 vaccine strains.…”