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Harvested virus suspension was pooled as a single batch and centrifuged at 5000 xg for 15 minutes at 4 C.Virus quantitation as mean tissue culture infective dose50 (TCID50) was calculated by the Spearman-KArber method .
The concentrated virus was mixed with the appropriate media and added to the wells to a final concentration of 5000 x TCID50. 100 U/mL of IFN[alpha] (PBL, InterferonSource New Jersey, USA) and SeV MOI 0.5 were used as controls.
Inactivation of foot and mouth disease virus: The FMDO virus suspension of known TCID50 was inactivated by binary ethylene imine (BEI) solution.
Virus titres determined by (TCID50) were [10.sup.8.1]/ml for UPM 1/14 and [10.sup.7.2]/ml for UPM 2/14.
Testing repeatability occurred on three consecutive days, using seven different samples in triplicate (a sample in LD concentration, three samples a log above LD and three samples 1/2 log above LD in the respective concentrations from, [10.sup.-1.5] TCID50 m[L.sup.-1], [10.sup.-1.0] TCID50 m[L.sup.-1] and [10.sup.-0.5] TCID50 m[L.sup.-1]).
TCID50 was calculated based on the ratio of the CPE positive wells to CPE negative wells (table 1) as described previously, (11) and titer of the propagated virus was [10.sup.-5.25] TCID50/unit [volume.sup.-1].
In addition, the produt has a detection limit of ~0.04 median tissue culture infective dose (TCID50) per reaction, meaning it can detect patients with active virus replications.
Dogs from inoculated groups were sedated with dexmedetomidine and then infected intranasally and intratracheally with direct deposition of either Ca/WY or Eq/CO [10.sup.7] 50% tissue culture infectious dose (TCID50) onto the nasal and tracheal respiratory epithelium.
dpe, days postexposure; dpi, days postinoculation; HAI, hemagglutination inhibition; TCID50, 50% tissue culture infective dose.
This technique involves a serial dilution of heat-inactivated serum followed by incubation with 100-200 TCID50 virus which leads to neutralization of the virus with antibodies.
Then, cells were infected with HSV-1 or 2 (10 TCID50) and incubated at 37 C in 5% CO2 air and observed every day up to three days for cell cytopathic effect (CPE) using a light microscope.
At 6 weeks of age, piglets in groups 1, 2, 3, and 4 were inoculated intramuscularly with 2 x [10.sup.5] tissue culture infective does (TCID50) QY1 virus of P5, P60, P80, and P100, respectively.
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