Two New Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction Assays with Unique Target Sites for the Specific and Sensitive Detection of Lineages 1 and 2 West Nile Virus Strains

Journal of Veterinary Diagnostic Investigation - Tập 22 Số 5 - Trang 748-753 - 2010
Martin Eiden1, Ariel Vina-Rodrı́guez1, Bernd Hoffmann2, Ute Ziegler1, Martin H. Groschup1
1Institute for Novel and Emerging Infectious Diseases (Eiden, Vina-Rodriguez, Ziegler, Groschup), Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany
2Institute for Diagnostic Virology (Hoffmann), Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany

Tóm tắt

Two novel 1-step real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays for the simultaneous detection of West Nile virus (WNV) lineage 1 and 2 strains were developed. Primers and the probe of assay 1 target the 5′-untranslated region (UTR), whereas the amplicon of assay 2 is located in the nonstructural region NS2A, which enables an unambiguous and independent WNV diagnosis based on 2 different amplicons. Both assays allow the detection of as few as 2–4 genome copies of WNV strains NY99, Uganda B956, Kunjin, and Sarafend (all cultured on Vero cells). A new synthetic RNA mutant of the 5′-UTR amplicon, which contains 6 twist inverted base-pair changes at the probe attachment site, was used as external calibrator control.

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Thông tin
Thông tin xuất bản

Journal of Veterinary Diagnostic Investigation

Tập 22 Số 5

748-753

Thông tin tác giả