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THE COMPARATIVE STUDY OF ACTIVITY TUBERCULINS Zavgorodniy A.I., Stegniy B.T., Bilushko V.V. National scientific center Institute of experimental and clinical veterinary medicine, ...

-- [ 2 ] --

2065-2070. - Print ISSN 0304-8608 2. / .., .. ; ϳ . .. . - .: , 2007. - 671 . - ISBN 978-5-9532-0301-2.

3. Lamb, R. A., Kolakofsky, D. Paramyxoviridae: The viruses and their replication. In Fields Virology, 2001 - pp.1305-1340. Edited by D.M. Knipe & P.M.

Howley. Philadelphia:Lippincott Williams & Wilkins. - DOI 10.1099/vir.0.81715-0

4. . . / . , // [ ]. - : 2007. - 456 . ISBN 5-9668-0016-2

5. .. : . . . : 16.00.04. - . 1999, - 175. 619.616.98 578.831.2.

6. Saiki, R. K., Gelfand, D. H., Stoffel,,S. Scharf, J., Higuchi, R., Horn, G. T., Mullis, K. B., Erlich, H. A. Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase. in: Science. 239.1988, 487491. - ISSN 0036-8075.

7. PCR protocols / edited by John M.S. Bartlett, David Stirling. --2nd ed. p. cm. -- (Methods in molecular biology ; v. 226). Includes bibliographical references and index. Humana Press Inc. - Totowa, New Jersey, 2003. - ISBN 0-89603-642-1.

8. Frisk, A. L., Knig, M., et al. Detection of Canine Distemper Virus Nucleoprotein RNA by Reverse Transcription-PCR Using Serum, Whole Blood, and Cerebrospinal Fluid from Dogs with Distemper. Journal of Clinical Microbiology, Nov. 1999. - pp.

36343643. - PMCID: PMC85712.

9. Wang et al.. Differentiation of canine distemper virus isolates in fur animals from various vaccine strains by reverse transcription-polymerase chain reaction-restriction fragment length polymorphism according to phylogenetic relations in china.

- Virology Journal, 2011, 8:85. - DOI: 10.1186/1743-422X-8-85.

DETERMINE THE SENSITIVITY AND SPECIFICITY OF THE DIAGNOSTIC TEST SYSTEM

FOR PCR IDENTIFICATION OF CANINE DISTEMPER VIRUS GENOME

Katsimon V.V. Karpulenko M.S., Golovko O.A.

State Scientific Control Institute of Biotechnology and strains of microorganisms, Kiev Objective: To develop a diagnostic test system for the identification of genomic PCR canine distemper virus in a biological materaly different origin.

Materials and Methods. Using computer programs were picked specific primers to conserved regions of the genome of canine distemper virus. PCR was performed on a thermocycler chotiroh channel. Detection of the reaction products was performed using ellektroforeza 1.5 % agarose gel. Results were evaluated after viewing the gel under ellekroforez on a transilluminator with UV light for the presence or absence of red-orange

2.

nucleic acid fragments of a particular size. The specificity of the amplified fragment was determined by its position relative to a standard marker fragments.

Results. Synthesized were 3 pairs of oligonucleotide primers of which article CDV F1 and CDV R2 [8];

CDV F3 and CDV R4 [9], as well as their own development CDV F5 i CDV R6. For the results of studies found satisfactory properties developed when the pair of primers the annealing temperature 58 C. Sensitivity was determined by dilution Virus febris contagiosae canis. Conducted validation developed primer pair in comparison with commercial PCR test system "Polichum" and serological test system CITO TEST CDV Ag. Primers were sensitive and specific.

Conclusions. 1. Primers were developed to detect canine distemper based polymerase chain reaction.

1,7

2. Optimal annealing temperature makes 58 C. 3. A test system has a high sensitivity, component 10 TCID50 Virus febris contagiosae canis. 4. Comparison of the developed test systems with commercial PCR test system and serological LHP system which resulted in it, at least not inferior to commercial.

Keywords: virus genome, diagnostics, system test, distemper.

619:579.843.96:636.4

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