Suda Yasuo, Nagatomo Mami, Yokoyama Risa, Ohzono Mami, Aoyama Kazue, Zhang Xu, Nakajima Kazuhiko, Murakami Naoki, Shinoda Tadashi, Hirota Tatsuhiko, Yanagihara Sae, Nishi Jun-Ichiro
Department of Chemistry, Biotechnology and Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University, 1-21-40 Kohrimoto, Kagoshima, Kagoshima 890-0065, Japan.
SUDx-Biotec Corporation, 1-42-1 Shiroyama, Kagoshima, Kagoshima 890-0013, Japan.
Biotechnol Rep (Amst). 2015 May 19;7:64-71. doi: 10.1016/j.btre.2015.05.004. eCollection 2015 Sep.
A highly sensitive and convenient method for detecting influenza virus was developed using modified end-point melt curve analysis of a RT-qPCR SYBR Green method and influenza virus-binding sugar chain-immobilized gold-nanoparticles (SGNP). Because SGNPs capture influenza viruses, the virus-SGNP complex was separated easily by centrifugation. Viral RNA was detected at very low concentrations, suggesting that SGNP increased sensitivity compared with standard methods. This method was applied to clinical studies. Influenza viruses were detected in saliva of patients or inpatients who had been considered influenza-free by a rapid diagnostic assay of nasal swabs. Furthermore, the method was applied to a human trial of prophylactic anti-influenza properties of yogurt containing L-92. The incidence of influenza viruses in saliva of the L-92 group was found to be significantly lower compared to the control group. Thus, this method was useful for monitoring the course of anti-influenza treatment or preventive measures against nosocomial infection.
利用逆转录定量聚合酶链反应（RT-qPCR）SYBR Green法的改良终点熔解曲线分析以及流感病毒结合糖链固定化金纳米颗粒（SGNP），开发了一种检测流感病毒的高灵敏度且便捷的方法。由于SGNP可捕获流感病毒，通过离心可轻松分离病毒-SGNP复合物。在极低浓度下即可检测到病毒RNA，这表明与标准方法相比，SGNP提高了检测灵敏度。该方法已应用于临床研究。在通过鼻拭子快速诊断检测被认为无流感的患者或住院患者的唾液中检测到了流感病毒。此外，该方法还应用于一项关于含L-92酸奶预防流感特性的人体试验。结果发现，与对照组相比，L-92组唾液中流感病毒的发生率显著更低。因此，该方法对于监测抗流感治疗过程或预防医院感染措施很有用。
