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Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples
Published on: June 30, 2018
Chemiluminescence analysis for HBV-DNA hybridization detection with magnetic nanoparticles based DNA extraction from
Nongyue He1, Fang Wang, Chao Ma
1State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, P. R. China.
Abstract:
Molecular detection of HBV has a significant impact on prognosis and therapy of the disease. In this paper, a sensitive nucleic acid detection method of HBV was established taking advantage of magnetic nanoparticles (MNPs), chemiluminescence (CL) and polymerase chain reaction (PCR). HBV-DNA was extracted from hepatitis B positive human blood samples using MNPs adsorption method and biotin was labeled on the DNA segment after base insertion of bintin-dUTP in PCR. The biotinylated DNA segment was captured by amino probe immobilized on carboxyl MNPs and was detected by the chemiluminescence system of alkaline phosphatase catalyzing 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy) phenyl-1, 2-dioxetane. Different concentrations of HBV-DNA were detected under the optimized experiment conditions and the relevant CL intensity were obtained, which provided a novel research or clinic diagnosis method for the quantification detection of HBV-DNA.
