TY - JOUR
T1 - Development of a dot blot neutralizing assay for HHV-6 and HHV-7 using specific monoclonal antibodies
AU - Tsukazaki, Takashi
AU - Yoshida, Mariko
AU - Namba, Hikaru
AU - Yamada, Masao
AU - Shimizu, Noshinobu
AU - Nii, Shiro
N1 - Funding Information:
We thank Drs. H. Watarai and Y. Muraki for their technical suggestions. We also thank Drs. K. Tabuchi and T. Yamagata for helpful discussions and encouragement. Part of this research project was supported by a grant-in-aid from the Ministry of Health and Welfare, Japan.
PY - 1998/8
Y1 - 1998/8
N2 - To elucidate further immune responses to human herpesviruses 6 and 7 (HHV-6 and -7), a neutralizing antibody assay was established for these viruses using a dot blot method. Three monoclonal antibodies against HHV-6 and 12 monoclonal antibodies against HHV-7 were developed and characterized by radio-immunoprecipitation. One monoclonal antibody which recognizes the 135 kDa late polypeptide of HHV-6 and several which recognize the 125 kDa late polypeptide of HHV-7 were selected to monitor virus growth by a dot blot antigen-detection method. The dot blot method was then used for the assay of HHV-6 and -7 neutralizing antibodies in human serum samples. The neutralization endpoints determined by the dot blot were comparable to those determined by immunofluorescence (IF). The neutralizing antibody titers appeared to correlate with the antibody titers determined by the indirect IF antibody test. The dot blot neutralization assay is easy to perform, is highly reproducible and objective when compared with the conventional methods based on cytopathology or IF for determining neutralization endpoints. Copyright (C) 1998 Elsevier Science B.V.
AB - To elucidate further immune responses to human herpesviruses 6 and 7 (HHV-6 and -7), a neutralizing antibody assay was established for these viruses using a dot blot method. Three monoclonal antibodies against HHV-6 and 12 monoclonal antibodies against HHV-7 were developed and characterized by radio-immunoprecipitation. One monoclonal antibody which recognizes the 135 kDa late polypeptide of HHV-6 and several which recognize the 125 kDa late polypeptide of HHV-7 were selected to monitor virus growth by a dot blot antigen-detection method. The dot blot method was then used for the assay of HHV-6 and -7 neutralizing antibodies in human serum samples. The neutralization endpoints determined by the dot blot were comparable to those determined by immunofluorescence (IF). The neutralizing antibody titers appeared to correlate with the antibody titers determined by the indirect IF antibody test. The dot blot neutralization assay is easy to perform, is highly reproducible and objective when compared with the conventional methods based on cytopathology or IF for determining neutralization endpoints. Copyright (C) 1998 Elsevier Science B.V.
KW - Dot blot assay
KW - Human herpesvirus 6
KW - Human herpesvirus 7
KW - Monoclonal antibodies
KW - Neutralizing antibody titer
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U2 - 10.1016/S0166-0934(98)00051-2
DO - 10.1016/S0166-0934(98)00051-2
M3 - Article
C2 - 9766885
AN - SCOPUS:0032145153
SN - 0166-0934
VL - 73
SP - 141
EP - 149
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 2
ER -