Nanocomposites Research Group, Department of Chemical Engineering, Anna University, Chennai, TN 600025, India.
Biosens Bioelectron. 2011 Jul 15;26(11):4624-7. doi: 10.1016/j.bios.2011.05.006. Epub 2011 May 11.
The zoonotic protozoan parasite Cryptosporidium parvum poses a significant risk to public health. Due to the low infectious dose of C. parvum, remarkably sensitive detection methods are required for water and food industries analysis. However PCR affirmed sensing method of the causative nucleic acid has numerous advantages, still criterion demands for simple techniques and expertise understanding to extinguish its routine use. In contrast, protein based immuno detecting techniques are simpler to perform by a commoner, but lack of sensitivity due to inadequate signal amplification. In this paper, we focused on the development of a mere sensitive immuno detection method by coupling anti-cyst antibody and alkaline phosphatase on gold nanoparticle for C. parvum is described. Outcome of the sensitivity in an immuno-dot blot assay detection is enhanced by 500 fold (using conventional method) and visually be able to detect up to 10 oocysts/mL with minimal processing period. Techniques reported in this paper substantiate the convenience of immuno-dot blot assay for the routine screening of C. parvum in water/environmental examines and most importantly, demonstrates the potential of a prototype development of simple and inexpensive diagnostic technique.
微小隐孢子虫是一种人畜共患的原生动物寄生虫，对公共健康构成重大威胁。由于微小隐孢子虫的感染剂量很低，因此需要对水和食品工业进行非常敏感的检测。然而，基于 PCR 的病原体核酸检测方法具有许多优点，但仍需要简单的技术和专业知识来消除其常规使用的障碍。相比之下，基于蛋白质的免疫检测技术更容易被普通大众操作，但由于信号放大不足，灵敏度较低。在本文中，我们专注于开发一种简单而敏感的免疫检测方法，方法是将抗隐孢子虫抗体和碱性磷酸酶偶联到金纳米颗粒上，用于检测微小隐孢子虫。通过免疫斑点印迹检测，灵敏度提高了 500 倍（使用传统方法），并且在最小的处理周期内，可以肉眼检测到低至 10 个卵囊/mL。本文报道的技术证明了免疫斑点印迹分析在水/环境检查中常规筛查微小隐孢子虫的便利性，最重要的是，展示了简单、廉价的诊断技术原型开发的潜力。
