Terminal deoxynucleotidyl transferase associated with split G-quadruplex/hemin deoxyribozyme amplification detection for various contaminants in milk based on pregnancy test strip platform.

Contaminant residue analysis in milk can provide essential assistance for safety quality and contamination level management of milk production, which is critical for safeguarding public health. In this study, the pregnancy test strip is employed to achieve multiple analytes detection based on the specific recognition of aptamer and terminal deoxynucleotidyl transferase associated with split G-quadruplex/hemin deoxyribozyme system. Through the subsequent enzyme catalyzed reaction, the detection signal can be further amplified to improve the sensitivity. The method does not need to assemble test strip, prepare and purify antibodies/haptens, nor design complex probe sequences. By coupling human chorionic gonadotrophin with DNA probes and combining magnetic separation technology, the targets can be determined via the test strip. Under the optimized conditions, the visual detection limits for mercury ion, bisphenol A, and penicillin are 1, 0.1 and 0.05 nM, respectively. The detection results show that the method displays good accuracy and practicability in spiked milk sample. The method presents a simple scheme, low cost as well as good design versatility, which demonstrates great application prospect for the sensitive, low-cost, and convenient detection of food matrices.