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ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)

 

Biotechnologia Acta V. 13, No 6, 2020
Р. 58-63, Bibliography 17, English
Universal Decimal Classification: 619:616.98-078:578.842.2:577.2.08:636.4
https://doi.org/10.15407/biotech13.06.058

DEVELOPMENT OF RECOMBINANT POSITIVE CONTROL FOR AFRICAN SWINE FEVER VIRUS PCR DETECTION

 M. Kit, O. Zlenko, O. Solodiankin, V. Bolotin, A.Gerilovych

National Scientific Center “Institute of Experimental and Clinical Veterinary Medicine” of the National Academy of Agrarian Sciences of Ukraine, Kharkiv

Recombinant plasmids containing target sequences are widely used as positive controls for PCR laboratory diagnostics. The aim of the study was development of recombinant positive control containing a fragment of B646L gene of African swine fever virus. The sequence of interest encodes targets of all the PCR assays for African swine fever laboratory diagnostics recommended by World Organisation for Animal Health. A plasmid containing 1763 bp insertion was cloned in E .coli DH5? strain. After purification, the plasmid ten-fold serial dulutions were used as a positive control while PRC testing. A minimal detectable copy number was 20 copies per reaction for both conventional and real-time PCR assays. The developed plasmid could be used as a safe and effective positive control while ASF laboratory diagnostics by PCR.

Key words: African swine fever virus, molecular cloning, PCR, positive recombinant control.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2020