Specifická magnetická izolace E6 HPV16 upravených magnetizovatelných částic spolu s PCR a elektrochemickou detekcí

摘要

The majority of carcinomas that were developed due to the infection with humanpapillomavirus (HPV) are caused by high-risk HPV types, HPV16 and HPV18. These HPV typescontain the E6 and E7 oncogenes, so the fast detection of these oncogenes is an important point toavoid the development of cancer. Many different HPV tests are available to detect the presence ofHPV in biological samples. The aim of this study was to design a fast and low cost method for HPVidentification employing magnetic isolation, polymerase chain reaction (PCR) and electrochemicaldetection. These assays were developed to detect the interactions between E6-HPV16 oncogeneand magnetizable particles (MPs) using commercial Dynabeads M-280 Streptavidin particles andlaboratory-synthesized “homemade” particles called MANs (MAN-37, MAN-127 and MAN-164). Theyields of PCR amplification of E6-HPV16 oncogene bound on the particles and after the elution fromthe particles were compared. A highest yield of E6-HPV16 DNA isolation was obtained with both MPsparticles commercial M-280 Streptavidin and MAN-37 due to reducing of the interferents comparedwith the standard PCR method. A biosensor employing the isolation of E6-HPV16 oncogene withMPs particles followed by its electrochemical detection can be a very effective technique for HPVidentification, providing simple, sensitive and cost-effective analysis.