Progress Made in the Rapid, Sensitive, Precise, and Visual Detection of Agricultural Biosafety Target Molecules

CopyFrom:生物所 PublishTime:2023-07-04 13:48:57 Hits: 【Font:Small large

The Agricultural Biosafety Evaluation Research Center of the Biotechnology Institute of SAAS has developed and constructed a variety of new, accurate, rapid, and visual detection technology systems based on nucleic acid molecular characteristics of key target traits of biological breeding materials, combined with nanomaterials, CRISPR and lateral-flow chromatography technology. The relevant research results were published in the international academic journals Food Science and Human Wellness ((JCR Q1, IF:8.022), Food Control (TOP 1, IF:6.652), andCurrent Research in Food Science (JCR Q1, IF:6.269).

As of 2021, the cumulative planting area of global genetically modified (GM) crops has reached 1.955 billion hectares worldwide, which is about 112 times more than that in 1996. The large-scale application of GM crops has aroused widespread social concern, and the market regulation of GM crops is crucial to ensuring food security and biosafety. Different GM organism regulatory policies have been developed by different countries and regions. The implementation of management policies requires technical support. Transgenic assays tend to be rapid, visual, and can be used in field-portable on-site testing settings.

1. A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops

In response to the current situation that the proportion of GM crops planted with insect resistant/herbicide-tolerant traits is increasing, a dual RPA-based strip detection system (named "Dual-RPA-LFD") was established targeting the major insect-resistant/herbicide-tolerant genes Cry1Ab/Ac and CP4 EPSPS. In this study, the dual RPA-based strip strategy was realized by primer labeling and primer optimization, and the detection results were characterized by control line (C line), test line 1 (T1 line), and test line 2 (T2 line). The whole analysis process could be completed within 20 min at 37℃. The results showed that the strip method can specifically identify Cry1Ab/Ac and CP4 EPSPS genes, not affected by other genes, and was suitable for random samples in the market and GM crops of different components, with good specificity and practicality. Meanwhile, plasmids containing Cry1Ab/Ac gene and CP4 EPSPS gene as low as 100 copies/µL (223 fg/mL) could be detected, indicating high sensitivity of the Dual-RPA, LFD strategy.