Development and evaluation of a rapid IgM capture ELISA (JEV-Chex) for the diagnosis of Japanese encephalitis

Ravi, V. ; Desai, A. ; Balaji, M. ; Apte, Madhuri P. ; Lakshman, Latha ; Subbakrishna, D. K. ; Sridharan, G. ; Dhole, T. N. ; Ravikumar, B. V. (2006) Development and evaluation of a rapid IgM capture ELISA (JEV-Chex) for the diagnosis of Japanese encephalitis Journal of Clinical Virology, 35 (4). pp. 429-434. ISSN 1386-6532

Full text not available from this repository.

Official URL: http://www.sciencedirect.com/science/article/pii/S...

Related URL: http://dx.doi.org/10.1016/j.jcv.2005.11.004

Abstract

Background The IgM capture ELISA has been the most widely used diagnostic method for Japanese encephalitis. However, the lack of availability of validated commercial kits as well as the short shelf life of the kit reagents has limited the use of this technique to very few centres in Asia. Objectives Development and evaluation of a rapid IgM capture ELISA (JEV Chex) in comparison to the conventional IgM capture ELISA. Produce key reagents such as cell culture derived JEV antigen and biotinylated monoclonal antibody which are stable at room temperature. Study design The conventional IgM capture ELISA was modified to reduce the total assay time and two key reagents used in the assay JEV antigen and biotinylated anti-JEV monoclonal antibodies were rendered stable at room temperature using a special procedure. A multi-centric evaluation of this rapid ELISA was carried out using well characterized stored CSF and serum samples. Long term stability of the key reagents was also assessed over a period of 6 months. Results The rapid IgM capture ELISA developed by us showed complete concordance with the results obtained using the conventional ELISA at all the three centres where it was evaluated. In addition, the stability studies carried out with the inactivated cell culture antigen and the biotinylated monoclonal antibodies stored at room temperature for a period of 180 days revealed that both these reagents yielded consistent optical density values in the ELISA. Conclusions The rapid ELISA format of the IgM capture ELISA (JEV-Chex) developed by us as well as the stability of reagents achieved by us in this study is what renders this rapid IgM capture ELISA very robust and user friendly since reagents can be stored at 4 °C by peripheral labs.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:IgM Capture ELISA; Japanese Encephalitis Virus; Stable Reagents
ID Code:63653
Deposited On:29 Sep 2011 06:29
Last Modified:29 Sep 2011 06:29

Repository Staff Only: item control page