Agricultural and Biosystems Engineering, University of Arizona, Tucson, Arizona, USA 85721.
J Biol Eng. 2011 Dec 5;5:16. doi: 10.1186/1754-1611-5-16.
In a globalized word, prevention of infectious diseases is a major challenge. Rapid detection of viable virus particles in water and other environmental samples is essential to public health risk assessment, homeland security and environmental protection. Current virus detection methods, especially assessing viral infectivity, are complex and time-consuming, making point-of-care detection a challenge. Faster, more sensitive, highly specific methods are needed to quantify potentially hazardous viral pathogens and to determine if suspected materials contain viable viral particles. Fourier transform infrared (FTIR) spectroscopy combined with cellular-based sensing, may offer a precise way to detect specific viruses. This approach utilizes infrared light to monitor changes in molecular components of cells by tracking changes in absorbance patterns produced following virus infection. In this work poliovirus (PV1) was used to evaluate the utility of FTIR spectroscopy with cell culture for rapid detection of infective virus particles.
Buffalo green monkey kidney (BGMK) cells infected with different virus titers were studied at 1 - 12 hours post-infection (h.p.i.). A partial least squares (PLS) regression method was used to analyze and model cellular responses to different infection titers and times post-infection. The model performs best at 8 h.p.i., resulting in an estimated root mean square error of cross validation (RMSECV) of 17 plaque forming units (PFU)/ml when using low titers of infection of 10 and 100 PFU/ml. Higher titers, from 103 to 106 PFU/ml, could also be reliably detected.
This approach to poliovirus detection and quantification using FTIR spectroscopy and cell culture could potentially be extended to compare biochemical cell responses to infection with different viruses. This virus detection method could feasibly be adapted to an automated scheme for use in areas such as water safety monitoring and medical diagnostics.
在全球化的世界中，预防传染病是一个重大挑战。快速检测水中和其他环境样本中的存活病毒颗粒对于公共卫生风险评估、国土安全和环境保护至关重要。目前的病毒检测方法，特别是评估病毒感染力，既复杂又耗时，使得即时检测具有挑战性。需要更快、更敏感、更特异的方法来定量潜在危险的病毒病原体，并确定可疑材料是否含有存活的病毒颗粒。傅里叶变换红外（FTIR）光谱结合基于细胞的传感，可能提供一种精确的方法来检测特定的病毒。这种方法利用红外光通过跟踪病毒感染后产生的吸光度模式变化来监测细胞内分子成分的变化。在这项工作中，使用脊髓灰质炎病毒（PV1）来评估 FTIR 光谱与细胞培养相结合用于快速检测感染性病毒颗粒的效用。
在感染后 1-12 小时（h.p.i.）研究了用不同病毒滴度感染的水牛绿猴肾（BGMK）细胞。使用偏最小二乘（PLS）回归方法分析和建模细胞对不同感染滴度和感染后时间的反应。该模型在 8 h.p.i.时表现最佳，使用 10 和 100 个 PFU/ml 的低感染滴度时，交叉验证的估计均方根误差（RMSECV）为 17 个噬菌斑形成单位（PFU）/ml。也可以可靠地检测到更高的滴度，从 103 到 106 PFU/ml。
使用 FTIR 光谱和细胞培养检测和定量脊髓灰质炎病毒的这种方法可能会扩展到比较不同病毒感染的生化细胞反应。这种病毒检测方法可以灵活地适应自动化方案，用于水安全监测和医疗诊断等领域。
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