title: Novel, real-time cell analysis for measuring viral cytopathogenesis and the efficacy of neutralizing antibodies to the 2009 influenza A (H1N1) virus.
abstract: A novel electronic cell sensor array technology, the real-time cell analysis (RTCA) system, was developed to monitor cell events. Unlike the conventional methods labeling the target cells with fluorescence, luminescence, or light absorption, the RTCA system allows for label-free detection of cell processes directly without the incorporation of labels. Here, we used this new format to measure the cytopathic effect (CPE) of the 2009 influenza A (H1N1) virus and the efficacy of neutralizing antibodies in human sera to this virus. The real-time dynamic monitoring of CPE was performed on MDCK cell cultures infected with the H1N1 virus, ranging from 5.50×10(2) to 5.50×10(7) copies/mL. The resulting CPE kinetic curves were automatically recorded and were both time and viral load dependent. The CPE kinetics were also distinguishable between different H1N1 stains, as the onset of CPE induced by the A/Shanghai/37T/2009 H1N1 virus was earlier than that of the A/Shanghai/143T/2009 H1N1 virus. Furthermore, inhibition of H1N1 virus-induced CPE in the presence of human specific anti-sera was detected and quantified using the RTCA system. Antibody titers determined using this new neutralization test correlated well with those obtained independently via the standard hemagglutination inhibition test. Taken together, this new CPE assay format provided label-free and high-throughput measurement of viral growth and the effect of neutralizing antibodies, illustrating its potential in influenza vaccine studies.
authors.affiliation: Pathogen Diagnosis and Biosafety Department, Shanghai Public Health Clinical Center Affiliated to Fudan University, Shanghai, China.
authors.forename: Di
authors.initials: D
authors.lastname: Tian
authors.forename: Wanju
authors.initials: W
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: He
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Song
authors.forename: Zhitong
authors.initials: Z
authors.lastname: Zhou
authors.forename: Min
authors.initials: M
authors.lastname: Zheng
authors.forename: Yunwen
authors.initials: Y
authors.lastname: Hu
mesh_terms: D000328:Adult; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral; D002452:Cell Count; D002460:Cell Line; D049109:Cell Proliferation; D003588:Cytopathogenic Effect, Viral / Q000276:immunology*; D004285:Dogs; D006385:Hemagglutination Inhibition Tests / Q000379:methods*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007251:Influenza, Human / Q000097:blood / Q000276:immunology / Q000821:virology; D009500:Neutralization Tests / Q000379:methods*; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism; D014611:Vaccination; D019562:Viral Load / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D014357:Trypsin