Rapid Visual LAMP Method for Detection of Genetically Modified Organisms

We developed a novel loop-mediated isothermal amplification(LAMP)method using DNA captured on polyacrylamide microparticles (PAMMPs)as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection ofgenetically modified organisms (GMOs). Here, DNA was extracted bya fast and cost-effective method using PAMMPs. Four LAMP primers weredesigned for the PAMMPs@DNA-LAMP method to detect the cauliflowermosaic virus 35S (CaMV35S) promotor in GMOs. We thus developed thismethod for rapid extraction of DNA (5-10 min) and fast amplificationof DNA within & SIM;30 min at a constant temperature of 63 & DEG;C.Moreover, the DNA captured by PAMMPs (PAMMPs@DNA) could be effectivelydetected by both conventional and quantitative PCR (qPCR) and LAMP.The PAMMPs@DNA-LAMP method was validated with high specificity, sensitivity,and performance for practical sample analysis. This assay detected0.01% target sequences, which had a high specificity like qPCR andbetter than the conventional PCR (cPCR). Furthermore, PAMMPs@DNA-LAMPwas successfully used to extract and detect DNA from food samplesof the major crops (soybean, maize, rice, etc.). In summary, a novelPAMMPs@DNA-LAMP assay has been developed, which has higher sensitivityand spends less time than the cPCR detection using the conventionalDNA extracted process. This method offers a novel approach for rapiddetection of GMOs in the field.

Automated summary

We developed a novel loop-mediated isothermal amplification (LAMP) method using DNA captured on polyacrylamide microparticles (PAMMPs) as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection of genetically modified organisms (GMOs). The PAMMPs@DNA-LAMP method allowed for rapid DNA extraction (5-10 min) and amplification (around 30 min) at a constant temperature of 63°C. The DNA captured by PAMMPs could be effectively detected by both conventional and quantitative PCR (qPCR) and LAMP.