关键词：流感病毒；医药；灵敏度检测；纳米粒子
摘 要：A sugar-chain-immobilized chip (named Sugar Chip, SC) and gold nano-particles (SGNP) were developed. Using these tools, nano-biotechnology methods were developed to discriminate the viral strains and to detect viruses high sensitively. Influenza viruses bind to neuraminic acid containing sugar-chains on the cell surface at the first stage of infection. The binding potency of type A influenza viruses was evaluated using surface plasmon resonance (SPR) imaging and SC immobilized with 6 kinds of sugar-chains containing N-acetylneuraminic acid and 2 kinds of sugar-chains with no N-acetylneuraminic acid. The relative binding potency of viruses to 8 kinds of sugar-chains immobilized on the SC varied even though the viruses were classified in the same serotype. Using 242 strains of influenza virus, we obtained 1976 data sets, created a database, and developed an algorithm based on the nearest neighbor with Euclidean distance to estimate the most similar strains according to the relative sugar-chain binding potency. Based on the database and the algorithm, we estimated the influenza virus strains most similar to the 2009 pandemic viruses (HlNlpdm2009) that were obtained from clinical isolates. In addition, based on the binding data, we found that the sugar-chain, heparin, binds to every virus, which allowed us to prepare heparin-immobilized gold nano-particles. Then, using these nano-particles, we established a novel technique for a super sensitive detection method of influenza viruses in combination with a real time quantitative polymerase chain reaction (real time qPCR). Using this method, we were able to detect influenza viruses that existed in the saliva of asymptomatic patients. Supportive clinical evaluations, during the 2011-2012 season, for influenza diagnosis were also obtained.