Development of Magnetic Nanoparticles Based Nucleic Acid Extraction Method and Application in Hepatitis C Virus Chemiluminescent Detection

The extraction of high quality nucleic acid is a primary factor in the genetic studies and development of biosensors based on nucleic acid testing. Hepatitis C virus (HCV) is an RNA virus and increasing number of HCV infected patients has grown the concern for the development of fast, inexpensive and reliable techniques for early disease detection. Nanotechnology has been applied for the development of detection methods which can be easily modified into automated systems. In this paper, a method for detection of HCV based on enzyme free magnetic nanoparticles (MNPs) extraction of nucleic acid and chemiluminescence has been described. The specific lysis buffer conditions helped nucleic acids to adsorb on the surface of MNPs. Different factors which might affect the nucleic acid purification were investigated. The quantity and quality parameters of nucleic acid were recorded using Nanodrop 1000 spectrophotometer. Reverse transcriptase polymerase chain reaction (RT-PCR) was carried out to verify that extracted nucleic acid retained high quality and could be used in the downstream applications. Agarose gel electrophoresis analysis confirmed the successful amplification of He la cells, Escherichia coli and HCV target sequences. The chemiluminescent detection of HCV was achieved by successively incubating the biotin labeled RT-PCR products with probe labeled MNPs and Streptavidin-modified alkaline phosphatase (SA-ALP). The results approved the successful chemiluminescent detection of HCV.