4 research outputs found
A novel modified-indirect ELISA based on spherical body protein 4 for detecting antibody during acute and long-term infections with diverse Babesia bovis strains
Cattle sera positive by the RAP-1-based cELISA but negative by the SBP4-based MI-ELISA and IFA had negative results by Western blot analysis, suggesting possible false positive results in the cELISA. A. Molecular weight marker (48 to 180 Kd), B. K42-#21, C. W31-#Y-3, D. W31-#Y-11, E. W31-#0-3, F. W31-#Y-9, G. W31-#0-9, H. W31-#Y-10, I. W31-#Y-15, J. P21-#224, K. positive control serum with a band at 75kd representing B. bovis RAP-1 protein, J. negative control serum. Figure S2. Technical difference between the modified indirect ELISA and conventional indirect ELISA using rGST-SBP4 was illustrated in this figure. (DOCX 645 kb
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Atomic force microscopy characterization of normal and cancer prostate cells
The cell mechanical properties in terms of the parameters adhesion, Young's, and dilation modulus and stiffness values for the prostate cell lines RWPE-1 (benign), RWPE-2 (nonmetastatic), DU145 (moderately metastatic from brain metastasis), and PC3 (highly metastatic from bone metastasis) have been determined by atomic force microscopy (AFM). A large number of force-distance approach curves were obtained to discern differences between the cell lines. Significant differences in all the parameters were observed between the cell lines as indicated by statistical analysis of the data. In general it was found that as cells transitioned from a normal to a highly metastatic phenotype they had higher elasticity and stiffness, and greater ability to dilate and bend. The densities of cell surface molecules and average interaction distances of these molecules and AFM cantilever tip were also determined. A long and short interaction distance with the cantilever tip at small and large indentations was observed. These interaction distances increased from normal to highly metastatic cells with a decrease in the corresponding molecular densities. Taken together these results indicate that the response of normal and cancer cells to nanoindentation forces are significantly different reflecting their phenotypes
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Improved diagnostic performance of a commercial Anaplasma antibody competitive enzyme-linked immunosorbent assay using recombinant major surface protein 5-glutathione S-transferase fusion protein as antigen
The current study tested the hypothesis that removal of maltose binding protein (MBP) from recombinant antigen used for plate coating would improve the specificity of a commercial Anaplasma antibody competitive enzyme-linked immunosorbent assay (cELISA). The number of 358 sera with significant MBP antibody binding (≥30%I) in Anaplasma-negative herds was 139 (38.8%) when tested using the recombinant major surface protein 5 (rMSP5)-MBP cELISA without MBP adsorption. All but 8 of the MBP binders were rendered negative (<30%I) using the commercial rMSP5-MBP cELISA with MBP adsorption, resulting in 97.8% specificity. This specificity was higher than some previous reports, so to improve the specificity of the commercial cELISA, a new recombinant antigen designated rMSP5-glutathione S-transferase (GST) was developed, eliminating MBP from the antigen and obviating the need for MBP adsorption. Using the rMSP5-GST cELISA, only 1 of 358 Anaplasma-negative sera, which included the 139 sera with significant (≥30%I) MBP binding in the rMSP5-MBP cELISA without MBP adsorption, was positive. This resulted in an improved diagnostic specificity of 99.7%. The rMSP5-GST cELISA without MBP adsorption had comparable analytical sensitivity to the rMSP5-MBP cELISA with MBP adsorption and had 100% diagnostic sensitivity when tested with 135 positive sera defined by nested polymerase chain reaction. Further, the rMSP5-GST cELISA resolved 103 false-positive reactions from selected sera with possible false-positive reactions obtained using the rMSP5-MBP cELISA with MBP adsorption and improved the resolution of 29 of 31 other sera. In summary, the rMSP5-GST cELISA was a faster and simpler assay with higher specificity, comparable sensitivity, and improved resolution in comparison with the rMSP5-MBP cELISA with MBP adsorption