Key result
The optimized Fluorescent Antibody Test in Lab-Tek slide chambers produced a virus titer of 10^7.5 TCID50/mL, showing 100% similarity to the traditional cover-slip culture method.
Why the study?
Classical assays for TCID 50 determination of classical swine fever virus are time-consuming, require huge reagents, and are unsuitable for large sample numbers.
Optimization of FAT in LAB TEK slide chambers provides an accurate, reliable, and less tedious method for titrating classical swine fever virus compared to traditional cover-slip cultures.
Simplified Lab-Tek FAT yields equivalent CSF titers; extends cover-slip methods but leaves routine high-throughput adoption open.
Determination of virus titer is the key factor for development and quality control of classical swine fever (CSF) cell culture vaccines.Like any other non-cytopathic effect producing viruses, classical swine fever virus titration is done by immunological methods e.g.Fluorescent Antibody Test (FAT).These classical assays for the determination of Tissue Culture Infective Dose 50 (TCID 50 ) are time-consuming and require huge reagents.Hence, they are not very suitable for handling a large number of samples.An accurate and reliable technique for TCID 50 determination by FAT in cover-slip culture was optimized for titration of classical swine fever.Optimization of FAT in LAB TEK slide chamber could do away the tedious manipulation of cover-slip culture without affecting its final outcome.This newly optimized method of titration has shown 100% similar result to that of one full proof titration method of CSF virus in PK-15 cells in terms of fluorescent positive cells in wells and their fluorescent intensity.
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Mahapatra et al. (2023) studied Classical Swine Fever Virus (CSFV). Fluorescent Antibody Test (FAT) in Lab-Tek slide chamber vs. FAT in 24-well plate cover-slip culture was evaluated on Virus titer (TCID50/mL) (100% similar result). The optimized Fluorescent Antibody Test in Lab-Tek slide chambers produced a virus titer of 10^7.5 TCID50/mL, showing 100% similarity to the traditional cover-slip culture method.
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