Design of a Test-System based on the Real-time PCR Assay for Identification of Ibaraki Virus Genome Fragment

Автор: E.V. Aronova, I.M. Kalabekov, G.S. Burmakina, S.Zh. Tsybanov, and A.V. Lunitsin

Страница: 70-75

Design of a Test-System based on the Real-time PCR Assay for Identification of Ibaraki Virus Genome Fragment

Biotekhnologiya, 2012, N 6, P. 70-75

UDC 619:616.98.578.823.1:577.2

Section:  “Metrology, Standartization, and Control”

E.V. Aronova ^*,  I.M. Kalabekov,  G.S. Burmakina,  S.Zh. Tsybanov,  and  A.V. Lunitsin

The All-Russian Research Institute for Veterinary Virology and Microbiology,  601120, Pokrov, Vladimirskaya oblast Russia

e-mail:  leia13@mail.ru

In the present study, a test-system based on the real-time polymerase chain reaction assay for the detection of a genome fragment of the Ibaraki virus (IBAV) in cell culture and clinical samples has been designed. A pair of primers complementary to a virus genome segment 10 region were created for the identification of the IBAV RNA. The latter from strains propagating in a cell culture and from clinical samples were detected by the suggested PCR-assay. No cross-reactions were observed when 2 serotypes of EHDV and 2 serotypes of BTV, closely related to IBAV orbiviruses, were tested. The sensitivity of the PCR assay was 1,5 lg TCID_50/ml, or 24 DNA copies per reaction.

Key words:  Ibaraki virus,  real-time polymerase chain reaction,  recombinant plasmid,  reverse transcription,  test-system for detection of Ibaraki virus RNA by real-time PCR.

The full English version of the article was published in “Biotechnology in Russia”, 2012, Issue 6, pp. 70-75 as .V. Aronova, I.M. Kalabekov, G.S. Burmakina, S.Zh. Tsybanov, A.V. Lunithin “Design of a Test-System based on Real-time PCR Assay for Identification of Ibaraki Virus Genome Fragment”.

It is contained at the Russian Scientific Electron Library website:  http://elibrary.ru/item.asp?id=22959639

30.04.2015, 1627 просмотров.