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Home » Archive » 2010

TDK conference 2010

Detection of porcine circovirus genome by polymerase chain reaction and isothermal amplification
Falus Dorottya Adrienn - year 5
SzIU, Faculty of Veterinary Science, Department of Microbiology and Infectious Diseases
Supervisors: Dr. Tamás Tuboly, Dr. Márta Lőrincz

Abstract:

Porcine circoviruses (PCV) are present worldwide causing serious economic losses in the pig industry. The losses are in part due to the clinical manifestations of the infection (either alone or as in form of co-infections) by the pathogenic form (PCV2) of the virus, collectively named as PCV associated diseases (PCVD), but the subclinical infections are also of great impact as PCV2 is well known for its immunosuppressive nature, decreasing the efficacy of regular vaccinations and increasing the pathogenicity of even facultatively pathogenic microorganisms. To our current knowledge PCV2 under natural conditions is present only in domestic pigs and wild boars, but recent studies indicated that rodents at infected pig herds may transmit the virus.

Viruses belonging into the PCV2 group show a remarkable genetic variability, yet the most widely used detection method is polymerase chain reaction (PCR), a very sensitive method but at the same time ignorant for divergent genomes, as it operates with specific primer sequences, so unusual genomes may remain undetected. There are however, several other possibilities already for the amplification of unknown genomes, one of these, an isothermal method, namely the rolling circle amplification (RCA) that is capable of synthesizing a number of copies from unknown circular templates using random hexamers.

The current study compared the sensitivity and efficiency of PCR and RCA, using DNA templates extracted from infected organs of domestic pigs and wild boars. According to the results of these experiments it was concluded that, although RCA had several advantages over PCR, still the sensitivity of the method was consequently one log below of that measured for PCR.



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