Fifty microliters of samples in serial dilutions (from 1:2 to 1:5

Fifty microliters of samples in serial dilutions (from 1:2 to 1:512) was prepared in a 96-cell plate. RV adjusted to 200 TCID50 in 50 μL of virus diluent (10% concentrated Hanks balanced salt solution, pH FRAX597 molecular weight 7.4) was added to the cell plate containing serially diluted serum. The mixture of antibody and virus was mixed and incubated at 37°C for 1 h. Then 100 μL of MA104 cells (used for virus infection) was added to the antibody-virus mixture and incubated in a 5% CO2 incubator at 37°C for 5 days. The overlay medium was then discarded, after which the wells were washed three times with sterile PBS, pH 7.4, and stained with 1% crystal violet solution. Differences in the number

of plaques formed between treatments were examined for the level of significance by ANOVA. Statistical analysis Statistical significance was determined using ANOVA, with a P value < 0.05 considered as significant. Acknowledgements This work was supported by grants from the National Science and Technology Foundation of China (No. 2006BAD06A07) and the Program for Innovative Research Team of NEAU (No. CXZ008). The authors wish to thank Jos Seegers for providing plasmid pPG611.1 and bacterial strain L. casei ATCC 393. References 1. Paul PS, Lyoo YS: Immunogens of rotaviruses. Vet Microbiol 1993, 37:299–317.PubMedCrossRef 2. Estes MK: Rotaviruses and their replication. Fields Virology 2001, 4:1747–1785. 3. Rosen I, Parwani AV, Lopez S, Flores J, Saif L: Serotypic

differentiation of rotaviruses in field samples from diarrheic pigs by using nucleic acid probes specific for porcine VP4 and human and porcine VP7 genes. J Clin Microbiol Tyrosine-protein kinase BLK 1994, 32:311–317.PubMed 4. Winiarczyk S, Paul PS, Mummidi NCT-501 concentration S, Panek R, Gradzki Z: Survey of porcine rotavirus G and P genotype in Poland and the United States using RT-PCR. J Vet Med 2002, 49:373–378.CrossRef 5. Gatti MS, Ferraz MM, Racz ML, de

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