The sensitivity and specificity of the ICS depend strongly on the antibodies used in the test strip. We used IgG3 conjugated with colloidal gold as the detector antibody and IgG4 as capture antibody in the test line. Both are core reagents, and they determine the specificity and sensitivity of the ICS strip. In previous research,we used the CP and CI of LMoV to construct a recombinant protein (CP-CI200), and produced the first antibody
We chose these proteins because the CP and CI of a potyvirus are involved in intercellular movement of virions and their abundance in infected cytoplasm is relatively high. Therefore, these are effective antigens to produce antibodies. Further more, western blotting analysis showed that IgG3 specifically reacted with the viral CP and CI of LMoV CI plays a complementary role in the reaction between viral antigens and polyclonal antibodies, and this may explain the 100%sensitivity of the ICS test compared with quadruplex RT-PCR. Additionally, to avoid the competing reaction with the first antibody,we used the second antibody (IgG4) with high specificity as the capture antibody, which may bind to different epitopes of LMoV antigen. This may contribute to the higher specificity of ICS test for LMoV.
The sensitivity and specificity of the ICS depend strongly on the antibodies used in the test strip. We used IgG3 conjugated with colloidal gold as the detector antibody and IgG4 as capture antibody in the test line. Both are core reagents, and they determine the specificity and sensitivity of the ICS strip. In previous research,we used the CP and CI of LMoV to construct a recombinant protein (CP-CI200), and produced the first antibodyWe chose these proteins because the CP and CI of a potyvirus are involved in intercellular movement of virions and their abundance in infected cytoplasm is relatively high. Therefore, these are effective antigens to produce antibodies. Further more, western blotting analysis showed that IgG3 specifically reacted with the viral CP and CI of LMoV CI plays a complementary role in the reaction between viral antigens and polyclonal antibodies, and this may explain the 100%sensitivity of the ICS test compared with quadruplex RT-PCR. Additionally, to avoid the competing reaction with the first antibody,we used the second antibody (IgG4) with high specificity as the capture antibody, which may bind to different epitopes of LMoV antigen. This may contribute to the higher specificity of ICS test for LMoV.
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