DOI: 10.36016/VM-2019-105-4

Veterinary Medicine: inter-departmental subject scientific collection. 2019. Issue 105. P. 20–25.

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DETECTION OF GENETIC MATERIAL OF PORCINE CIRCOVIRUS TYPE II BY LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP)

Rudova N. G., Solodiankin O. S., Gerilovych A. P.

National Scientific Center “Institute of Experimental and Clinical Veterinary Medicine”, Kharkiv, Ukraine

There is a sufficient number of molecular-genetic methods for the Porcine Circovirus type II (PCV-II) detection, based on conventional or real-time PCR. However, these methods require expensive equipment, heat cycles for amplification, and considerable time to perform the study. The aim of our work was to develop an alternative method of the PCV-II detection based on isothermal amplification (LAMP), which characterized by cost-effectiveness and short time of study performing. By this reaction a few copies of DNA to 10⁹ molecules might be amplified in about one hour at a constant temperature which is suitable for the field conditions. We designed a set of primers using the target cap gene sequence with the further parameters optimizing of the amplification protocol. Amplification was performed for 60 minutes in a water bath, and the result was observed in UV light using a transilluminator by the adding SYBR green I to the reaction mixture. The elaborated set of primers for LAMP showed high sensitivity and specificity. The set of primers was designed to take into account the molecular genetic features of PCV, and it can significantly expand the range of existing molecular genetic screening techniques for PCV –II detection

Keywords: PCV-II, loop-mediated isothermal amplification, set of primers

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