A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses

被引：14
|
作者：
Wang, Heng [1 ]
Nattanmai, Seela [1 ]
Kramer, Laura D. [1 ,2 ]
Bernard, Kristen A. [1 ,2 ]
Tavakoli, Norma P. [1 ,2 ]
机构：
[1] Wadsworth Ctr, New York State Dept Hlth, Albany, NY 12201 USA
[2] SUNY Albany, Sch Publ Hlth, Dept Biomed Sci, Albany, NY USA
来源：
DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2009年 / 65卷 / 02期
基金：
美国国家卫生研究院;
关键词：
Molecular detection; Real-time RT-PCR; California serogroup viruses; Cache Valley virus; LA-CROSSE ENCEPHALITIS; RNA SEGMENT; BUNYAMWERA; BUNYAVIRIDAE; INFECTIONS; AMPLIFICATION; ARBOVIRUSES; PREVALENCE; SEQUENCES; DIAGNOSIS;
D O I：
10.1016/j.diagmicrobio.2009.07.001
中图分类号：
R51 [传染病];
学科分类号：
100401 ;
摘要：
A duplex TaqMan real-time reverse transcriptase polymerase chain reaction (PCR) assay was developed for the detection of California (CAL) serogroup viruses and Cache Valley virus (CVV), for use in human surveillance. The targets selected for the assay were the sequences encoding the nucleocapsid protein of CAL and the G I glycoprotein of CVV. Conserved regions were selected by aligning genetic sequences from various strains available in the GenBank database. Primers and probes were selected in conserved regions. The assay sensitivity was 75 gene copies (gc)/reaction for CAL serogroup viruses and 30 gc/reaction for CVV. The performance of the assay was linear over at least 6 log(10) gc. The assay was specific, given that it did not cross-react with a variety of pathogens. It did, however, detect 11 viruses within the CAL serogroup and 12 CVV isolates. The use of an internal control ensured that possible inefficiency in nucleic acid extraction or PCR inhibition would be detected. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码：150 / 157
页数：8
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