title: A simple fluorescent labeling technique to study virus adsorption in Newcastle disease virus infected cells.
abstract: The present study demonstrates that the fluorescent general membrane dyes PKH67 and PKH26 are suitable to label Newcastle disease virus, an enveloped virus belonging to the family of paramyxoviridae. Adsorption of the labeled virus particles was tracked, visualized and quantitated using confocal laser scanning microscopy. The specificity of PKH-labeling was determined by colocalization analysis of the PKH signal with NDV-specific immunolabeling, and by using mock-infected controls and infection with detergent-pretreated labeled virus particles. The infectivity of the NDV particles was not affected by the labeling procedure as indicated by the results of a cytotoxicity ATP assay, an apoptosis assay and detection of virus-specific RNA and protein by qPCR and Western blotting, respectively, in cells infected with PKH-labeled and unlabeled virus particles. This technique can be used as an inexpensive, sensitive and rapid alternative method in the analysis of adsorption and internalization of enveloped viruses by the infected cells.
authors: Department of Medical Biology, Medical School, University of Pécs, Szigeti 12., H-7624 Pécs, Hungary.
authors: András
authors: A
authors: Balogh
authors: Marianna
authors: M
authors: Pap
authors: Lajos
authors: L
authors: Markó
authors: Ibolya
authors: I
authors: Koloszár
authors: László K
authors: LK
authors: Csatáry
authors: József
authors: J
authors: Szeberényi
mesh_terms: D000327:Adsorption; D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor / Q000821:virology; D002462:Cell Membrane / Q000648:ultrastructure / Q000821:virology; D003588:Cytopathogenic Effect, Viral; D003593:Cytoplasm / Q000821:virology; D003902:Detergents / Q000494:pharmacology; D004705:Endocytosis; D019085:Fluorescent Antibody Technique, Direct; D005456:Fluorescent Dyes / Q000032:analysis*; D008561:Membrane Fusion; D018613:Microscopy, Confocal; D009522:Newcastle disease virus / Q000187:drug effects / Q000502:physiology*; D009930:Organic Chemicals / Q000032:analysis*; D010673:Pheochromocytoma / Q000473:pathology; D012367:RNA, Viral / Q000032:analysis; D051381:Rats; D013194:Staining and Labeling / Q000379:methods*; D053586:Virus Internalization
chemical_list: D003902:Detergents; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26; C451241:PKH67; D012367:RNA, Viral