Based on intervening PCR for detection of alkaline phosphatase and zearalenone

Abstract

Polymerase chain reaction (PCR) is an excellent technique for amplifying DNA. It is worth studying to expand the application of PCR to detect other targets besides DNA. In this work, a dNTP hydrolysis mediated PCR was used to detect alkaline phosphatase (ALP) and mycotoxin zearalenone (ZEN). With dephosphorylation, the ALP can hydrolyze dNTP to inhibit the PCR. Thus, the PCR signal was negatively correlated with the concentration of ALP. By combining PCR and enzyme-linked immunosorbent assay (ELISA), targets can be extended further. Represented by ZEN, a dNTP hydrolysis mediated PCR-ELISA for detecting ZEN was presented. The ZEN was associated with ALP-labelled goat anti-mouse IgG (IgG-ALP) by indirect competitive ELISA. Subsequently, IgG-ALP can interfere with the PCR. This proposed method did not require the preparation of raw materials such as nanomaterials. For ALP detection, the linear range was 10–125 U/L with limit of detection (LOD) of 0.93 U/L. In dNTP hydrolysis mediated PCR-ELISA, the linear range for detecting ZEN was 6–14 ng/mL with LOD of 0.168 ng/mL.