Abstract
A reliable and sensitive direct competitive real-time immuno-PCR (rt-IPCR) assay was developed for the determination of coplanar 3,3′,4,4′-tetrachlorobiphenyl (PCB 77). PCB77 butanoic acid (γ-oxo-PCB77A) was synthesized as the hapten of PCB77, and a mixed anhydride reaction was used to couple the PCB77A to ovalbumin to form artificial coating antigen. The active ester method was used to couple the PCB77A to bovine serum albumin to form the artificial antigen. Male New Zealand white rabbits were immunized with immune antigen to obtain polyclonal antibodies (pAbs), with which, a novel rt-IPCR assay for determination of PCB77 was described. Using the optimized assay, the linear range for the determination of PCB77 is 10.0 fg mL−1 to 1 ng mL−1 with a correlation coefficient of 0.98 and a detection limit of 1.5 fg mL−1. The rt-IPCRs were tested for their cross-reactivity profiles using 4 selected congeners and 4 Aroclor products. The assays were highly specific for congeners but less specific for Aroclor1248. The rt-IPCR results for soil samples correlated well with the concentrations of PCBs obtained by GC/MS. Recovery was lower or higher with agitation but would still be acceptable for use in an on-site field test to provide rapid, semi-quantitative, and reliable test results for making environmental decisions.
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