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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.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2019: 96.39
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39
For more details click here

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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 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2021.10(6): 188-197
DOI: https://doi.org/10.20546/ijcmas.2021.1006.020


Evaluation of Single Step TaqMan Real-time PCR Assay Lateral to Conventional RT-PCR and Antigen-Capture ELISA for Pre-Clinical Detection of Classical swine fever virus
Elina Khatoon1, 2, Mousumi Bora1, 3, Gitika Rajbongshi1, 4,Seema Rani Pegu5 and Nagendra Nath Barman1*
1Department of Microbiology, College of Veterinary Science, Guwahati, Assam-781022, India
2Department of Biosciences and Bioengineering, Indian Institute of Technology, Guwahati, Assam-781039, India
3Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, Banaras Hindu University, Uttar Pradesh-231001, India
4Department of Microbiology, Gauhati Medical College, Guwahati, Assam, India
5Department of Animal Health, National Research Centre on Pigs, Rani, Assam-781129, India
*Corresponding author
Abstract:

Classical swine fever (CSF) is a highly contagious and devastating viral disease, causing serious losses in the pig industry worldwide. Rapid detection and identification of the causative agent is a crucial step in controlling CSF infection in pig population. In the present study, a fluorogenic-probe hydrolysis (TaqMan)-reverse transcriptase real time PCR assay (RT-qPCR)was evaluated parallel to conventional RT-PCR and antigen capture ELISA to detect Classical swine fever virus (CSFV) in the pre-clinical phase of the disease. In addition, hematological analysis was performed at different clinical phases in order to diagnose CSF pre-clinically. Thrombocytopenia and leucopenia were early clinical clues recorded in CSFV infected pigs. Single step RT-qPCR confirmed the presence of CSFV nucleic acid in blood, nasal swabs, ocular swabs as well as in tonsillar scrapings in the pre-clinical phase. CSFV nucleic acid was detected with maximum positivity in blood and tonsillar scrapings (70-73%) using RT-qPCR as compared to 60% and 33.33-40% positivity in conventional RT-PCR and Ag-ELISA, respectively. Thus, TaqMan based RT-qPCR assay can be used as an efficient assay for rapid CSFV detection at pre-clinical phase of the disease to contain the disease from in-contact infected pigs to susceptible population.


Keywords: Classical swine fever virus, pre-clinical detection, TaqMan RT-qPCR, RT-PCR, antigen capture-ELISA
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How to cite this article:

Elina Khatoon, Mousumi Bora, Gitika Rajbongshi, Seema Rani Pegu and Nagendra Nath Barman. 2021. Evaluation of Single Step TaqMan Real-time PCR Assay Lateral to Conventional RT-PCR and Antigen-Capture ELISA for Pre-Clinical Detection of Classical swine fever virus.Int.J.Curr.Microbiol.App.Sci. 10(6): 188-197. doi: https://doi.org/10.20546/ijcmas.2021.1006.020