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- Title
Comparison of RT-PCR assay and virus isolation in cell culture for the detection of alkhumra hemorrhagic fever virus.
- Authors
Madani, Tariq A.; Abuelzein, El‐Tayb M.E.; Azhar, Esam I.; Al‐Bar, Hussein M.S.; Abu‐Araki, Huda; Ksiazek, Thomas G.
- Abstract
Alkhumra hemorrhagic fever virus (AHFV) is an emerging flavivirus that was isolated originally from Saudi Arabia in 1994-1995. The main tests used for the detection of AHFV are the real time (rt) RT-PCR and virus isolation in cell culture. In the present study the detection of AHFV by rtRT-PCR was compared with virus isolation in BHK-21, HEp-2, and LLC-MK2 cell lines. AHFV suspensions grown in BHK-21, HEp-2, and LLC-MK2 cell lines were serially diluted 10-fold from 10−1 to 10−11. Samples from each dilution were used to inoculate four cell culture tubes and were also examined by the rtRT-PCR for AHFV RNA. Fifteen non-inoculated cell culture samples (five from each cell line) were included blindly in both tests. Thus, a total of 132 AHFV-positive and 15 negative control samples were tested. The rtRT-PCR could detect the viral RNA in all diluted specimens up to and including the 10−10 dilution (40 specimens for each cell line), whereas, cell cultures were positive in 70% of specimens for BHK-21, 65% for LLC-MK2, and 45% for HEp-2 at this dilution. None of the three cell cultures nor the rtRT-PCR was positive at 10−11 dilution. The specificity and positive predictive values of virus isolation compared to rtRT-PCR were each 100%, whereas the negative predictive values were 29.4% for BHK-21, 26.3% for LLC-MK2, and 18.5% for HEp-2. In conclusion, the rtRT-PCR is more sensitive than virus isolation for detecting AHFV. J. Med. Virol. 86:1176-1180, 2014. © 2013 Wiley Periodicals, Inc.
- Publication
Journal of Medical Virology, 2014, Vol 86, Issue 7, p1176
- ISSN
0146-6615
- Publication type
Article
- DOI
10.1002/jmv.23755