Quantification of Mycobacterium avium subsp. paratuberculosis from the tissues of challenged mice using SYBR Green real time PCR assay for the assessment of vaccine efficacy
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Date
2019-07
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Abstract
A SYBR Green real time PCR assay amplifying F57 gene of Mycobacterium avium subsp. paratuberculosis (MAP) was
developed to evaluate the protective efficacy of the inactivated vaccine to prevent the colonization of MAP in the tissues
of BALB/c mice challenge model. Bacterial burden in the liver, spleen and intestine of vaccinated and sham immunized
control mice, after intra peritoneal challenge with MAP were quantified by real time PCR assay. A 195 bp fragment of
MAP F57 sequence was cloned into TA cloning vector and the resultant recombinant plasmid was used to generate a series
of quantification standards with copy number ranging from 109 to 1 copy. This absolute quantification technique is rapid
and able to detect as few as 10 MAP organisms per gram of tissue and the assay has the efficiency of 0.982.
Description
TNV_IJAR_2019_53(7)944-948
Keywords
Veterinary Science