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- Title
Comparison of virus isolation using the Vero E6 cellline with real-time RT-PCR assay for the detectionof human metapneumovirus.
- Authors
Matsuzaki, Yoko; Mizuta, Katsumi; Takashita, Emi; Okamoto, Michiko; Itagaki, Tsutomu; Katsushima, Fumio; Katsushima, Yuriko; Nagai, Yukio; Nishimura, Hidekazu
- Abstract
Background: The use of cell culture for the diagnosis of human metapneumovirus (hMPV) infection is uncommon at present and molecular method such as reverse-transcription PCR (RT-PCR) has been widely and most commonly used as the preferred test. We aimed to compare the results of virus isolation using Vero E6 cells with real-time RT-PCR for the detection of hMPV, since such a comparison data is not available. Methods: Between December 2007 and July 2008, we obtained 224 nasopharyngeal swab specimens from patients with acute respiratory infection and tested by the two methods. Results: Forty-three (19.2%) were found positive by cell culture and 62 (27.7%) by real-time RT-PCR. Cell cultures were positive for 42 of 62 specimens found positive by real-time RT-PCR (67.7% sensitivity) and for 1 of 162 specimens found negative by real-time RT-PCR (99.4% specificity), respectively. The sensitivity of the cell culture was 76.2-87.5% (mean 81.8%) when specimens were collected within 3 days after the onset of symptoms, and the sensitivity decreased to 50% or less thereafter. Among specimens collected within 3 days after symptom onset, all of the real-time RT-PCR positive specimens having a viral load of more than 1.25×105 copies/ml were found positive by cell culture. Conclusions: Cell culture using Vero E6 cell line has 81.8% sensitivity compared with the real-time RT-PCR method, when specimens are collected within 3 days after the onset of symptoms. Thus, this method is a useful method for epidemiological and virological research even in facilities with minimal laboratory resources.
- Subjects
CELL culture; VIRUS isolation; REVERSE transcriptase polymerase chain reaction; SARS disease; EPIDEMIOLOGICAL research
- Publication
BMC Infectious Diseases, 2010, Vol 10, Issue 1, p170
- ISSN
1471-2334
- Publication type
Article
- DOI
10.1186/1471-2334-10-170