2014.ISDS.Abstracts.Final.pdf


ISDS Annual Conference Proceedings 2014. This is an Open Access article distributed under the terms of the Creative Commons Attribution-
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ISDS 2014 Conference Abstracts

Development of the Multiplex PCR for Detection of the 
DNA-Contained Emergent Diseases Agents in Pigs 
(African Swine Fever, Aujeszky Disease, Circoviral 
Disease)
Anton Gerilovych*, Prof. Borys Stegniy, Prof. Valeriy Golovko, Dr Oleksii Solodiankin, Dr 
Iryna Gerilovych, Yevgenia Smolyaninova and Natalya Rudova

NSC IECVM, Kharkiv, Ukraine

Objective
The objective was to develope the multiplex PCR for detection of 

the DNA-contained emergent diseases agents in pigs (African swine 
fever, Aujeszky disease, Circoviral disease) for diseases surveilance 
in pig farms

Introduction
African swine fever virus (ASFV), pseudorabies virus (Aujrszky’s 

disease virus, ADV) and porcine circovirus type 2 (PCV-2) are 
some of the most dangerous DNA viruses causing high amounts 
of morbidity and mortality in commercial and backyard pig farms. 
Traditional diagnosis of porcine viral infections requires complex and 
lengthy combinations of virological and serological tests. This study 
aimed to develop a method for rapid detection of the DNA-containing 
viral pathogens of pig in clinical materials using conventional 
multiplex PCR platform.

Methods
A PCR-based detection technique for ASFV, ADV and PCV-2 

detection has been developed and tested under Office International 
Epizootical (O.I.E.) requirements, using determination of specificity, 
sensitivity and repeatability.

Results
The optimal conditions for amplification of ASFV, ADV and 

PCV-2 genome in p72, gE and cap genes, respectively, were 
determined. The developed protocol allowed to detect ASFV DNA by 
276 bp fragment, Aujeszky disease virus and PCV-2 – 230 and 421 bp, 
respectively. Validation testing was performed on the panel selected 
agents: ASFV recombinant DNA, cloned in pUC19, ADV strain 
UNDIEV18V, and PCV isolates 1-02, 6-12, and 4-12. The protocol 
demonstrated specificity in its ability to distinguishbetween Porcine 
parvovirus, Classical swine fever virus, and Porcine Respiratory and 
Reproductive Syndrome virus. The protocol was 90 % sensitive, 100 
% specific and repeatable. It was recommended for validation and 
implementation in the laboratory practice in Ukraine.

Conclusions
A multiplex PCR platform for detection of the emergent disease 

agents in pigs (African swine fever, Aujeszky disease, Circoviral 
disease) has been developed for preliminary diagnostics of these 
diseases.

Keywords
multiplex PCR; porcine viroses; african swine fever

*Anton Gerilovych
E-mail: antger@rambler.ru    

Online Journal of Public Health Informatics * ISSN 1947-2579 * http://ojphi.org * (1):e131, 201