Calculating rabies virus neutralizing antibodies titres by flow cytometry
Rev. Inst. Med. Trop. Säo Paulo
;
44(3): 151-154, 2002. graf
Article
in English
| LILACS, SES-SP
| ID: lil-314543
RESUMO
The determination of the rabies neutralizing antibody (VNA) response after immunization against rabies is an acceptable index of the efficacy of a vaccine and a successful treatment. Several tests have been developed in attempt to improve the assessment of VNA, from mice inoculation to cell-culture fluorescence inhibition tests. All of them, however, present special difficulties in terms of reading or accuracy. The present study describes a neutralization test performed in cell-culture appraised by flow cytometry (FC). Serial dilutions of the serum samples were mixed in vitro with rabies virus before the addition of BHK-21 cells. After 24h-incubation, cells were released by trypsin treatment, fixed and permeabilized with a p-formaldehyde solution and stained with a rabies virus nucleocapsid protein-specific antibody conjugate. The percentage of virus infection inhibition caused by specific antibodies present in the serum were evaluated in a Beckton & Dickinson FACSCalibur® flow cytometer. A correlation curve between the IU/ml content and the percentage of infective inhibition was built with a reference serum and the VNA titers of serum samples were obtained by extrapolation. Titers obtained by FC and standard test showed an effective pairing results (p < 0.01), with a correlation coefficient (r) = 0.7. These results permit to envisage the FC as a suitable technique to evaluate VNA in sera from immunized animals and likely in human serum samples. Nevertheless, new studies comparing FC to gold-standard techniques are required for determining the FC values of Sensibility and Specificity
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Rabies virus
/
Rabies Vaccines
/
Flow Cytometry
/
Antibodies, Viral
Type of study:
Diagnostic study
Limits:
Animals
Language:
English
Journal:
Rev. Inst. Med. Trop. Säo Paulo
Year:
2002
Type:
Article
Institution/Affiliation country:
Center of Hemotherapy of Santa Catarina/BR
/
Federal University of Santa Catarina/BR
/
Pasteur Institute/BR
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