Application of the pseudo-plaque assay for detection and titration of Crimean-Congo hemorrhagic fever virus.
A pseudo-plaque assay was developed for detection and quantitation of Crimean-Congo hemorrhagic fever virus Turkey-Kelkit06. Enzyme-catalyzed color development of infected cells probed with anti-Crimean-Congo hemorrhagic fever virus antibodies was used for determining the titer of Crimean-Congo hemorrhagic fever Turkey-Kelkit06 and for its detection in samples from persons infected with the Crimean-Congo hemorrhagic fever virus. The pseudo-plaque assay accuracy was confirmed by comparing pseudo-plaque assay titers with fluorescent immunofocus assay and focus formation assay titers using three stocks of virus. No significant difference in virus titers of Crimean-Congo hemorrhagic fever Turkey-Kelkit06 among the three methods was observed. The pseudo-plaque assay is more sensitive than the fluorescent immunofocus assay for detecting the virus in primary isolates of Crimean-Congo hemorrhagic fever virus collected from humans, but no difference in sensitivity between the two methods was observed in the cell-adapted strain of Crimean-Congo hemorrhagic fever Turkey-Kelkit06. The pseudo-plaque assay is suitable for titration of Crimean-Congo hemorrhagic fever Turkey-Kelkit06, which does not develop plaques, suggesting it may also be suitable for the detection of other viruses.
Department of Virology, College of Veterinary Medicine, Firat University, Elazig, Turkey.
Engin
E
Berber
Nurettin
N
Canakoglu
Mustafa D
MD
Yoruk
Sukru
S
Tonbak
Munir
M
Aktas
Mustafa
M
Ertek
Yusuf
Y
Bolat
Ahmet
A
Kalkan
Aykut
A
Ozdarendeli
D000914:Antibodies, Viral; D004797:Enzyme-Linked Immunosorbent Assay; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000276:immunology* / Q000302:isolation & purification; D006479:Hemorrhagic Fever, Crimean / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D006801:Humans; D012680:Sensitivity and Specificity; D010948:Viral Plaque Assay
D000914:Antibodies, Viral