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

Full Abstract

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 G1 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.

Author information

Author/s: Wang, Heng (H); Nattanmai, Seela (S); Kramer, Laura D (LD); Bernard, Kristen A (KA); Tavakoli, Norma P (NP);

Affiliation: Wadsworth Center, New York State Department of Health, Albany, NY 12201, USA.

Grants: N01-A1-25490 (Agency:PHS HHS)

Journal and publication information

Publication Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

Journal: Diagnostic microbiology and infectious disease (Diagn Microbiol Infect Dis), published in United States. (Language: eng)

Reference: 2009-Oct; vol 65 (issue 2) : pp 150-7

Dates: Created 2009/09/14; Completed 2009/11/06; Revised 2009/11/09;

PMID: 19748425, status: MEDLINE (last retrieval date: 11/10/2009, IMS Date: )

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