Follow
International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00 For more details click here
National Academy of Agricultural Sciences (NAAS) : NAAS Score: *5.38 (2019) [Effective from January 1, 2019]For more details click here

Login as a Reviewer

Indexed in



National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

See Guidelines to Authors
Current Issues

Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2019.8(2): 1693-1698
DOI: https://doi.org/10.20546/ijcmas.2019.802.198


Detection of Corynebacterium pseudotuberculosis by SYBR Green Real Time PCR
Jyoti Kumar1*, G.G. Sonawane1, S. Jegaveera Pandianand Rajiv Kumar2
1Division of Animal Health
2 Division of Animal Genetics and Breeding, ICAR-Central Sheep and Wool Research Institute, Avikanagar 304501, India
*Corresponding author
Abstract:

Corynebacterium pseudotuberculosis, a Gram positive bacterium is the causative agent of caseous lymphadenitis (CLA), a chronic debilitating disease of sheep, goats and other warm blooded animals. Definite diagnosis of CLA primarily relies on microbiological examination, followed by biochemical identification of isolates. In an effort to facilitate C. pseudotuberculosis detection, the present study was undertaken with the aim of evaluating a SYBR Green real time PCR assay targeting ABC transporter gene for the detection of C. pseudotuberculosis in DNA isolated from culture and pus samples from sheep and goats. The analytical sensitivity and specificity of the PCR primers were evaluated. The test demonstrated tenfold more sensitivity than conventional PCR and detected down to 100 femtogram of genomic DNA of pure C. pseudotuberculosis. The real-time PCR was found to be specific for ABC transporter gene of C. pseudotuberculosis, as no cross reactivity was detected with a variety of known bacterial isolates. The real time PCR was employed for direct detection of C. pseudotuberculosis in pus samples collected from clinically suspected cases of caseous lymphadenitis. It is concluded that, this real time assay may be used as a valuable diagnostic tool for the rapid and specific detection of C. pseudotuberculosis in clinical samples, large scale epidemiological studies and clarification of pathogenesis.


Keywords: Real time PCR, Corynebacterium pseudotuberculosis, sensitivity, specificity, SYBR green
Download this article as Download

How to cite this article:

Jyoti Kumar, G.G. Sonawane, S. Jegaveera Pandian and Rajiv Kumar. 2019. Detection of Corynebacterium pseudotuberculosis by SYBR Green Real Time PCR.Int.J.Curr.Microbiol.App.Sci. 8(2): 1693-1698. doi: https://doi.org/10.20546/ijcmas.2019.802.198