Fluorometric cell-ELISA for quantifying rabies infection and heparin inhibition.
The purpose of the present study was to implement a fluorometric method for detecting and quantifying viral antigens in human meduloblastoma cells infected by two types of fixed rabies virus (CVS-MB and CVS-BHK) and a street virus using a cell-enzyme linked immunosorbent assay (cell-ELISA) technique; alkaline phosphatase was used as the antibody-marker enzyme and 4-methyl-umbelliferyl-phosphate as the fluorogenic substrate. The system was used for detecting up to 1:10,000 viral inoculums, followed by evaluating the effect of heparin on infection. Infected cultures were reliably differentiated from their respective negative controls in both assays allowing data to be analysed statistically. As reported in another study, heparin produces strong inhibition when the CVS-BHK viral strain is used for infection; it has thus been suggested that it binds to the neural cell adhesion molecule and could be blocked by using this drug. This fluorometric method is less time-consuming, has increased reproducibility and useful for quantitation of collected data and can therefore be considered as a useful tool for research.
Rincón
Verónica
V
Laboratorio de Neurociencias, Instituto Nacional de Salud, Bogotá, Colombia.
Corredor
Adriana
A
Martínez-Gutíerrez
Marlén
M
Castellanos
Jaime E
JE
D000918:Antibody Specificity; D000956:Antigens, Viral / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005470:Fluorometry; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives; D019006:Neural Cell Adhesion Molecules / Q000096:biosynthesis / Q000737:chemistry; D011820:Rabies virus / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification*
D000956:Antigens, Viral; D019006:Neural Cell Adhesion Molecules; C005359:4-methylumbelliferyl phosphate; D006923:Hymecromone; D006493:Heparin