Evaluation of variables in immunofluorescence procedures for the detection of antibodies against human herpesvirus 6 (HHV-6).
Ten human sera were used to study different parameters, namely, methods of smear preparation and fixation, and age of infected HSB-2 cells in order to optimize indirect immunofluorescence assay (IFA) and anticomplement immunofluorescence (ACIF) procedures to measure antibody levels against HHV-6. Results showed a greater sensitivity of rapid smear drying and methanol fixation over conventional acetone smear preparation. Cells harvested 6 days after infection and fixed with methanol exhibited a sharper and more intense fluorescence. IFA titers were higher than those obtained with ACIF, although the latter procedure enabled the distinction between three fluorescent sites. Reactivity pattern of individual sera against infected cells was variable and indicated that the human immune response to HHV-6 is directed against different antigens. An easier interpretation and a better definition of the fluorescence of HSB-2 cell line infected with HHV-6 strain Dv is obtained with the following conditions: cells should be harvested at 5-8 days after infection (at the giant cell stage of infection), cell smears have to be dried quickly before fixation with methanol at -20 degrees C, and finally, they should be stained by IFA.
Couillard
M
M
Department of Microbiology, Faculty of Medicine, Laval University, Sainte-Foy, Québec, Canada.
Joly
J R
JR
Deschênes
L
L
Richer
G
G
D000096:Acetone; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D003165:Complement System Proteins / Q000276:immunology; D005455:Fluorescent Antibody Technique; D015654:Herpesvirus 6, Human / Q000276:immunology / Q000648:ultrastructure*; D016591:Histocytological Preparation Techniques; D006801:Humans; D000432:Methanol; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D000096:Acetone; D003165:Complement System Proteins; D000432:Methanol