Isolation of RNA from blood and RT-PCR for detection of live Brucella organisms in cattle v1

Raj, Sugandha; Mohan, Hari

doi:10.17504/protocols.io.mayc2fw

2017

DOI: 10.17504/protocols.io.mayc2fw

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Isolation of RNA from blood and RT-PCR for detection of live Brucella organisms in cattle v1

Sugandha Raj¹,

Hari Mohan²

Abstract: A protocol for isolation of RNA from blood and RT-PCR for detection of live Brucella organisms in cattle

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A novel immunotherapy of Brucellosis in cows monitored non invasively through a specific biomarker

Saxena

Raj

2018

PLoS Negl Trop Dis

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Brucellosis is an important zoonotic disease causing huge economic losses worldwide. Currently no effective immunotherapy for Brucellosis or any biomarker to monitor the efficacy of therapy is available. Treatment is ineffective and animals remain carrier lifelong. S19 and RB51 are live attenuated vaccine strains of Brucella abortus. However, S19 induces only antibody, ineffective for intracellular pathogen. RB51 induces cell mediated immunity (CMI) but it is Rifampicin resistant. Both organisms are secreted in milk and can infect humans and cause abortions in animals. Phage lysed bacteria (lysates) retain maximum immunogenicity as opposed to killing by heat or chemicals. We report here the successful immunotherapy of bovine Brucellosis by phage lysates of RB51 (RL) and S19 (SL). The SL induced strong antibody response and RL stimulated CMI. In vitro restimulation of leukocytes from RL immunized cattle induced interferon gamma production. A single subcutaneous dose of 2 ml of cocktail lysate (both RL and SL), eliminated live virulent Brucella from Brucellosis affected cattle with plasma level of Brucella specific 223 bp amplicon undetectable by RT-PCR and blood negative for live Brucella by culture in 3 months post-immunization. This is the first report on minimally invasive monitoring of the efficacy of antibacterial therapy employing plasma RNA specific for live bacteria as a biomarker as well as on the use of RB51 phage lysate for successful immunotherapy of Brucellosis in cattle.

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A novel immunotherapy of Brucellosis in cows monitored non invasively through a specific biomarker

Saxena

Raj

2018

PLoS Negl Trop Dis

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Isolation of RNA from blood and RT-PCR for detection of live Brucella organisms in cattle v1

Abstract: A protocol for isolation of RNA from blood and RT-PCR for detection of live Brucella organisms in cattle

Cited by 1 publication

References 0 publications

A novel immunotherapy of Brucellosis in cows monitored non invasively through a specific biomarker

A novel immunotherapy of Brucellosis in cows monitored non invasively through a specific biomarker

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