Enhanced Bio-Barcode Immunoassay Using Droplet Digital PCR for Multiplex Detection of Organophosphate Pesticides


Cui X., Abd El-Aty A. M., Zhang C., Xu L., Liu H., Jia H., ...More

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, vol.69, no.37, pp.11131-11141, 2021 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 69 Issue: 37
  • Publication Date: 2021
  • Doi Number: 10.1021/acs.jafc.1c03216
  • Journal Name: JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Analytical Abstracts, Applied Science & Technology Source, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Chimica, Compendex, EMBASE, Environment Index, Food Science & Technology Abstracts, MEDLINE, Pollution Abstracts, Veterinary Science Database, DIALNET
  • Page Numbers: pp.11131-11141
  • Keywords: gold nanoparticles, magnetic nanoparticles, oligonucleotides, ddPCR, organophosphate pesticides, RESIDUE DETECTION, AMPLIFICATION
  • Ataturk University Affiliated: Yes

Abstract

A bio-barcode immunoassay based on droplet digital polymerase chain reaction (ddPCR) was developed to simultaneously quantify triazophos, parathion, and chlorpyrifos in apple, cucumber, cabbage, and pear. Three gold nanoparticle (AuNP) probes and magnetic nanoparticle (MNP) probes were prepared, binding through their antibodies with the three pesticides in the same tube. Three groups of primers, probes, templates, and three antibodies were designed to ensure the specificity of the method. Under the optimal conditions, the detection limits (expressed as IC10) of triazophos, parathion, and chlorpyrifos were 0.22, 0.45, and 4.49 ng mL(-1), respectively. The linear ranges were 0.01-20, 0.1-100, and 0.1-500 ng mL(-1), and the correlation coefficients (R-2) were 0.9661, 0.9834, and 0.9612, respectively. The recoveries and relative standard deviations (RSDs) were in the ranges of 75.5-98.9 and 8.3-16.7%. This study provides the first insights into the ddPCR for the determination of organophosphate pesticides. It also laid the foundation for high-throughput detection of other small molecules.