Chemidoc mp imaging system machine

The protein A/G-based ICT assay [2 (link)] was used to detect the anti-P. insidiosum antibodies in 16 recruited serum samples of humans and animals with or without pythiosis (Serum IDs: RbCFA, HuP1–3, HsP1–3, HsB1–3, HuC1, and HsC1–5) (Table 2). Briefly, an ICT strip was dipped into 100 µL of a diluted serum (1:5000 (animal serum) and 1:10,000 (human serum) in PBS). The antibodies formed a complex with the colloidal gold-conjugated protein A/G and diffused through the ICT strip. The anti-P. insidiosum antibodies, if present, bound the pre-blotted PiCFA and generated a visualized signal at the test line (Figure 2). The remaining colloidal gold-protein A/G-antibody complex passed through the test line, captured the pre-blotted rabbit anti-IgG antibodies, and generated a visualized signal at the control line (Figure 2). The ICT result was read within 30 min by experienced personnel. The presence of both the test and control lines was read ICT positive, while the presence of the only control line was read ICT negative. Each tested strip was photocopied using a ChemiDoc MP Imaging System machine (Bio-Rad, Hercules, CA, USA).