Sharma Anshul, Kaushal Ankur, Kulshrestha Saurabh
Faculty of Applied Sciences and Biotechnology, Shoolini University of Biotechnology and Management Sciences, Bajhol, Solan, Himachal Pradesh, 173229, India.
Department of Food Science and Biotechnology, Gachon University, Seongnam, 13120, South Korea.
Arch Virol. 2017 Jul;162(7):2047-2052. doi: 10.1007/s00705-017-3293-5. Epub 2017 Mar 14.
Accurate and on time diagnosis of plant viruses is an essential prerequisite for efficient control in field conditions. A number of diagnostic methods have been reported with the required level of sensitivity. Here, we propose a label free immunosensor for efficient and sensitive detection of capsicum chlorosis virus (CaCV) in bell pepper. Antigen was immobilized over the surface of gold nanoparticle/multi-walled carbon nanotube (Nano-Au/C-MWCNT) screen printed electrodes using 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) cross linking chemistry followed by interaction with groundnut bud necrosis virus (GBNV)/CaCV specific polyclonal antibody. The electrochemical response was measured by cyclic voltammetry (CV), differential pulse voltammetry (DPV) using the redox indicator. Electrode surface characterization was done by performing scanning electron microscopy (SEM). Electrochemical studies showed positive results at different antigenic dilutions ranging from 10 - 8x10. The sensitivity of the immunosensor developed has been compared with direct antigen coated enzyme-linked immunosorbent assay (DAC-ELISA) and the results showed that the immunosensor developed was 800-1000 times more sensitive, when compared to DAC-ELISA for CaCV detection. The immunosensor we have developed is economical and sensitive and could be used for immediate determination of the presence of virus in extracts from bell pepper leaves.
在田间条件下，准确及时地诊断植物病毒是有效防控的必要前提。已经报道了许多具有所需灵敏度水平的诊断方法。在此，我们提出一种无标记免疫传感器，用于高效灵敏地检测甜椒中的辣椒褪绿病毒（CaCV）。使用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺（EDC）/N-羟基琥珀酰亚胺（NHS）交联化学方法，将抗原固定在金纳米颗粒/多壁碳纳米管（Nano-Au/C-MWCNT）丝网印刷电极表面，随后与花生芽坏死病毒（GBNV）/CaCV特异性多克隆抗体相互作用。使用氧化还原指示剂通过循环伏安法（CV）、差分脉冲伏安法（DPV）测量电化学响应。通过扫描电子显微镜（SEM）对电极表面进行表征。电化学研究表明，在10 - 8×10的不同抗原稀释度下均得到阳性结果。将所开发的免疫传感器的灵敏度与直接抗原包被酶联免疫吸附测定（DAC-ELISA）进行了比较，结果表明，在所开发的免疫传感器用于检测CaCV时，其灵敏度比DAC-ELISA高800 - 1000倍。我们所开发的免疫传感器经济且灵敏，可用于立即测定甜椒叶片提取物中病毒的存在情况。
