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Enhancing the Detection of Giardia duodenalis Cysts in Foods by Inertial Microfluidic Separation

机译:惯性微流控分离增强食品中贾第鞭毛虫囊肿的检测

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The sensitivity and specificity of current Giardia cyst detection methods for foods are largely determined by the effectiveness of the elution, separation, and concentration methods used. The aim of these methods is to produce a final suspension with an adequate concentration of Giardia cysts for detection and a low concentration of interfering food debris. In the present study, a microfluidic device, which makes use of inertial separation, was designed and fabricated for the separation of Giardia cysts. A cyclical pumping platform and protocol was developed to concentrate 10-ml suspensions down to less than 1 ml. Tests involving Giardia duodenalis cysts and 1.90-μm microbeads in pure suspensions demonstrated the specificity of the microfluidic chip for cysts over smaller nonspecific particles. As the suspension cycled through the chip, a large number of beads were removed (70%) and the majority of the cysts were concentrated (82%). Subsequently, the microfluidic inertial separation chip was integrated into a method for the detection of G. duodenalis cysts from lettuce samples. The method greatly reduced the concentration of background debris in the final suspensions (10-fold reduction) in comparison to that obtained by a conventional method. The method also recovered an average of 68.4% of cysts from 25-g lettuce samples and had a limit of detection (LOD) of 38 cysts. While the recovery of cysts by inertial separation was slightly lower, and the LOD slightly higher, than with the conventional method, the sample analysis time was greatly reduced, as there were far fewer background food particles interfering with the detection of cysts by immunofluorescence microscopy.
机译:当前对食品进行的贾第虫囊肿检测方法的敏感性和特异性在很大程度上取决于所使用的洗脱,分离和浓缩方法的有效性。这些方法的目的是生产一种最终悬浮液,该悬浮液应具有足够浓度的贾第虫囊肿,以进行检测,并提供低浓度的干扰性食物残渣。在本研究中,设计并制造了一种利用惯性分离的微流体装置来分离贾第虫囊肿。开发了循环泵送平台和方案以将10 ml悬浮液浓缩至不足1 ml。在纯悬液中进行的贾第鞭毛虫囊肿和1.90-μm微珠试验表明,微流控芯片对囊肿的特异性优于较小的非特异性颗粒。随着悬浮液在芯片中循环,大量珠子被去除(70%),大部分囊肿被浓缩(82%)。随后,将微流惯性分离芯片整合到一种从生菜样品中检测十二指肠囊肿的方法中。与常规方法相比,该方法大大降低了最终悬浮液中背景碎片的浓度(减少了10倍)。该方法还从25克生菜样品中平均回收了68.4%的囊肿,检出限(LOD)为38个囊肿。与传统方法相比,通过惯性分离得到的囊肿回收率略低,LOD略高,但由于干扰了免疫荧光显微镜检测的囊肿的背景食品颗粒少得多,因此样品分析时间大大缩短。

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