Department of Virology, College of Veterinary Medicine, Firat University, Elazig, Turkey.
J Virol Methods. 2013 Jan;187(1):26-31. doi: 10.1016/j.jviromet.2012.07.025. Epub 2012 Aug 10.
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.
建立了一种用于检测和定量分析克里米亚-刚果出血热病毒土耳其-凯利特 06 株的假蚀斑法。用抗克里米亚-刚果出血热病毒抗体探测感染细胞的酶促显色反应可用于确定克里米亚-刚果出血热土耳其-凯利特 06 株的效价，并用于检测感染该病毒的人的样本中的病毒。通过比较三种病毒株的假蚀斑法效价与荧光免疫焦点法和空斑形成法效价，证实了假蚀斑法的准确性。在三种方法中，未观察到克里米亚-刚果出血热土耳其-凯利特 06 株病毒效价的显著差异。假蚀斑法比荧光免疫焦点法更敏感，可用于检测从人类中分离的克里米亚-刚果出血热病毒的原始分离物，但在细胞适应的克里米亚-刚果出血热土耳其-凯利特 06 株中，两种方法的敏感性没有差异。假蚀斑法适用于效价滴定克里米亚-刚果出血热土耳其-凯利特 06 株，因为它不能形成蚀斑，这表明它也可能适用于其他病毒的检测。
