Selected article for: "Culture infectious dose and infectious dose"

Author: Thomas, Joseph T.; Chen, Qi; Gauger, Phillip C.; Giménez-Lirola, Luis G.; Sinha, Avanti; Harmon, Karen M.; Madson, Darin M.; Burrough, Eric R.; Magstadt, Drew R.; Salzbrenner, Holly M.; Welch, Michael W.; Yoon, Kyoung-Jin; Zimmerman, Jeffrey J.; Zhang, Jianqiang
Title: Effect of Porcine Epidemic Diarrhea Virus Infectious Doses on Infection Outcomes in Naïve Conventional Neonatal and Weaned Pigs
  • Document date: 2015_10_6
  • ID: 1kjk404o_8
    Snippet: The virus stock and serial dilutions were back titrated in Vero cells to determine their infectious titers in the unit of median tissue culture infective dose per milliliter (TCID 50 /ml) and plaque forming unit per milliliter (PFU/ml). To determine TCID 50 /ml, each sample was serially 10-fold diluted in post-inoculation media and inoculated into Vero cells grown in 96-well plates, 100μl per well, triplicate wells per dilution. The plates were .....
    Document: The virus stock and serial dilutions were back titrated in Vero cells to determine their infectious titers in the unit of median tissue culture infective dose per milliliter (TCID 50 /ml) and plaque forming unit per milliliter (PFU/ml). To determine TCID 50 /ml, each sample was serially 10-fold diluted in post-inoculation media and inoculated into Vero cells grown in 96-well plates, 100μl per well, triplicate wells per dilution. The plates were incubated at 37°C with 5% CO 2 for 5 days. Viral cytopathic effects (CPE) were recorded daily. After 5-day inoculation, the plates were subjected to immunofluorescence staining using a monoclonal antibody conjugate SD6-29 against the PEDV nucleocapsid protein (SD-1F-1, Medgene Labs, Brookings, South Dakota, USA). The virus titers were determined according to the method described by Reed and Muench [23] and expressed as TCID 50 /ml. To determine PFU/ml, each sample was serially 10-fold diluted in post-inoculation media and inoculated into Vero cells grown in 6-well plates, 500μl per well, duplicate wells per dilution. After incubation at 37°C for 1 h, 4ml of overlay medium (post-inoculation media supplemented with 0.75% carboxymethyl cellulose) was added to each well. The plates were then incubated undisturbed at 37°C with 5% CO 2 for 4 days. The media were discarded and the plates were stained with crystal violet in 10% buffered formalin. The bottoms of the plates were then illuminated. Wells containing 20-200 plaques were counted and titers expressed as PFU/ml.

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