Real-time replication of swine vesicular disease virus (SVDV) in cell culture systems in vitro

Open access


A swine vesicular disease virus (SVDV) replication assay in IB-RS-2, SK-6, and PK-15 cell cultures was performed using the xCELLigence system. The cell status was monitored by impedance measurement, expressed as cell index (CI). Proliferation of particular cells was examined at the beginning of the study. The cells exhibited the ability to form a monolayer, and the CI values increased with the cell culture growth. After about 23 h and while still in the growth phase, the cells were infected with decimal virus dilutions (10-1-10-6) containing from 100 000 to 1 median tissue culture infectious doses (TCID50). SVDV replication in cell cultures induced a change in cell index; together with the occurrence of cytopathic effect (CPE), the CI values declined. A significant correlation between the concentration of the virus used and CPE occurrence was found. The results also enabled determination of cell sensitivity to SVDV infection. The highest sensitivity was exhibited by IB-RS-2, followed by SK-6. To conclude, the xCELLigence System was used effectively and evaluated as being an efficient tool for CPE detection and SVDV replication analysis in cell cultures. Compared to the standard method , it enabled a more precise assessment of viral replication based on the quantitative CI measurement, providing additional current information.

1. Escribano-Romero E., Jimenez-Clavero M.A., Ley V.: Swine vesicular disease virus. Pathology of the disease and molecular characteristics of the virion. Anim Health Res Rev 2000, 1, 119-126.

2. Fang Y., Ye P., Wang X., Xu X., Reisen W.: Real-time monitoring of flavivirus induced cytopathogenesis using cell electric impedance technology. J Virol Methods 2011, 173, 251-258.

3. Golke A., Cymerys J., Słońska A., Dzieciątkowski T., Chmielewska A., Tucholska A., Bańbura M.W.: The xCELLigence system for real-time and label-free analysis of neuronal and dermal cell response to equine herpesvirus type 1 infection. Pol J Vet Sci 2012, 15, 151-153.

4. Huang B., Jin D., Zhang J., Sun J.Y., Wang X., Stiles J., Xu X., Kamboj M., Babady N.E., Tang Y.W.: Real-time cellular analysis coupled with a specimen enrichment accurately detects and quantifies Clostridium difficile toxins in stool. J Clin Microbiol 2014, 52, 1105-1111.

5. Irelan J.T., Wu M.-J., Morgan J., Ke N., Xi B., Wang X., Xu X., Abassi Y.A.: Rapid and quantitative assessment of cell quality, identity, and functionality for cell-based assays using real-time cellular analysis. J Biomol Screen 2011, 16, 313-322.

6. Lin F., Kitching R.P.: Swine Vesicular Disease: An Overview. Vet J 2000, 160, 192-201.

7. Paprocka G.: Detection of swine vesicular disease virus in biological materials. Med Weter 2010, 2, 118-120.

8. Paprocka G., Niedbalski W., Kęsy A., Fitzner A.: Detection of foot-and-mouth disease virus in biological samples - recent diagnostics possibilities. Bull Vet Inst Pulawy 2010, 54, 467-472.

9. Ryder A.B., Huang Y., Li H., Zheng M., Wang X., Stratton C.W., Xu X., Tang Y.W.: Assessment of Clostridium difficile infections by quantitative detection of tcdB toxin by use of a real-time cell analysis system. J Clin Microbiol 2010, 48, 4129-4134.

10. Slanina H., König A., Claus H., Frosch M., Schubert- Unkmeir A.: Real-time impedance analysis of host cell response to meningococcal infection. J Microbiol Methods 2011, 84, 101-108.

11. Solly K., Wang X., Xu X., Strulovici B., Zheng W.: Application of real-time cell electronic sensing (RT-CES) technology to cellbased assays. Assay Drug Dev Technol 2004, 2, 363-372.

12. Tian D., Zhang W., He J., Liu Y., Song Z., Zhou Z., Zheng M., Hu Y.: Novel, real-time cell analysis for measuring viral cytopathogenesis and the efficacy of neutralizing antibodies to the 2009 influenza A (H1N1) virus. PLoS ONE 2012, 7, e31965.

13. Witkowski P.T., Schuenadel L., Wiethaus J., Bourquain D.R., Kurth A., Nitsche A.: Cellular impedance measurement as a new tool for poxvirus titration, antibody neutralization testing and evaluation of antiviral substances. Biochem Biophys Res Commun 2010, 401, 37-41.

14. Teng Z., Kuang X., Wang J., Zhang X.: Real-time cell analysis - a new method for dynamic, quantitative measurement of infectious viruses and antiserum neutralizing activity. J Virol Methods 2013, 193, 364-370.

Bulletin of the Veterinary Institute in Pulawy

The Journal of National Veterinary Research Institute in Pulawy

Journal Information

IMPACT FACTOR 2017: 0.811
5-year IMPACT FACTOR: 0.973

CiteScore 2017: 0.68

SCImago Journal Rank (SJR) 2017: 0.287
Source Normalized Impact per Paper (SNIP) 2017: 0.484


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 111 111 14
PDF Downloads 40 40 5