Detection Of Genetically Modified Foods And Genotyping HPV Infection By A GeXP Based Multiplex PCR Assay

Objective:To establish a GeXP based multiplex PCR assay, and evaluate its preliminary application in detection of genetically modified components in foods and genotyping human papillomavirus.Methods:A GeXP based multiplex PCR assay was developed to detect simultaneously six transgenes (PAT,NOS,CaMV35S,FMV35S,Bar,EPSPS) from genetically modified soybean and genetically modified rapeseed meal. The detection results were compared with those of the real-time PCR protocol for detecting genetically modified plants and their derived products documented as the industry standard by the State Administration for Entry-Exit Inspection and Quarantine.A GeXP based multiplex PCR assay was developed to genotype human papillomavirus (HPV) 6,11,31,33 and 52, which are the most common subtypes in Chinese population. The specificity and sensitivity of this assay were evaluated and the detection results were compared with that of a commercial kit.Resμlts:We successfully developed a GeXP based multiplex PCR assay to detect simultaneously six genetically modified components (PAT,NOS,CaMV35S, FMV35S,Bar,EPSPS) from soybean and rapeseed meal samples. In this assay, 100ng of PCR template of each target was used to meet the industry standard required by the State Administration for Entry-Exit Inspection and Quarantine. The detection results were consistent with those deduced from the real-time PCR protocol.We also successfully established a GeXP based multiplex PCR assay to detect simultaneously five HPV subtypes which are the most common types in Chinese population(including two low-risk types HPV6,HPV11 and three high-risk types HPV31,HPV33, HPV52). The assay achieved a sensitivity of 103 copies of DNA per reaction with high specificity. The detection results of 30 clinical samples were consistent with those obtained from a commercial kit. Conclusion:We have uccessfully established a GeXP based multiplex PCR platform, and preliminarily applied in the detection of genetically modified components in foods and genotyping of human papillomavirus, which provides a new idea and method for achieving high throughput detection of multiple target genes.