Wang M.-S.,Changchun BCHT Biopharm Co. |
Zheng Y.,Changchun BCHT Biopharm Co. |
Guo L.-S.,Changchun BCHT Biopharm Co. |
Tag L.,Changchun BCHT Biopharm Co. |
And 2 more authors.
Chinese Journal of Biologicals | Year: 2014
Objective To optimize and verify the condition for determination of rabies virus titer by direct immunofluorescence assay (dFA). Methods The influences of inoculation method (stepwise and one-step inoculations) as well as temperature (34 and 37 °C) and time (6, 12, 24, 48, 72 and 96 h) on titer of rallies viius were evaluated bv direct immunofluorescence assay. The optimized method was verified for inter-precision and specificity, and the results were compared with those of intracerebral titration in mice. Results Hie cells were inoculated by one-step inoculation, while vims was cultured at 34 °C for 96 h and stained to evaluate the test result. Fluorescent foci were counted on dav 3 after virus culture to calculate the virus titer. The variation coefficient of 12 test results by two personnel was only 0. 05%. Specific fluorescence was observed onlv in rabies vims but not in canine distemper vims or canine parvovirus by the method. The determination results of virus titers by dFA showed no significant difference with that by intracerebral titration in mice (P> 0. 05 ). Conclusion The optimized dFA is precise, specific, easy to handle and time-saving, which may be used for the quantitative determination of rabies vims liter.