A rapid one-step immunochromatographic test strip for rabies detection using canine serum samples

Abstract

Significance and Impact of the Study: Simple and cheap techniques to detect rabies virus or monitor immunity against it are central in maintaining epidemiological control over the disease, particularly in endemic developing countries. While many techniques meet this requirement, they are confined to this usage as they are time-consuming and demand expensive instrumentation. Our immunochromatographic test strip can detect rabies antibody with high specificity and sensitivity; the output can be measured with naked eye. It allows safe and quick detection that will be of value in the surveillance of the immunization status of potential targets in rabies-endemic regions and will aid disease control. AbstractWe developed an immunochromatographic test strip using colloidal gold-coated staphylococcal protein A (SPA) for the detection of rabies antibody in canine serum samples. The recombinantly expressed rabies virus phosphoprotein (RV-P) and the anti-staphylococcal protein A (anti-SPA) polyclonal antibody were coated on the test (T) and control (C) lines on a nitrocellulose membrane, respectively. This layout is designed such that the polyclonal antibody in canine serum is captured by the colloidal gold-SPA conjugates, before the rabies antibody complex is specifically selected by the RV-P deposited on the T line, forming a 'sandwich' pattern. Unbound excess colloidal SPA then proceeds to the control line where SPA specifically interacts with the anti-SPA antibody, producing a red precipitation at the C line, indicating the validity of the strip. We tested 165 canine serum samples with the strips, and the results were compared with those obtained using ELISA. The specificity and sensitivity of ICTS were found to be 93Á1 and 92Á2%, respectively. As a rapid technique, not demanding expensive instrumentation, the strip offers potential in disease monitoring, especially in rabies-endemic developing countries.

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

“…Virus isolation and real-time PCR assays are reliable methods for diagnosis of CAdV infections; however, these methods are mostly restricted to well-equipped laboratories. The ICS method has been widely applied to detect various infectious agents (Li et al, 2011;Nakayama et al, 2014;Liu et al, 2019), including canine viruses such as rabies and CPV (Tao and Li, 2014;Sharma et al, 2018). However, there are no publications regarding ICS detection of CAdV, although there is commercial ICS that detect CAdV-2 on the market.…”

Rabies – epidemiology, pathogenesis, public health concerns and advances in diagnosis and control: a comprehensive review

“…For the diagnosis of RABV specific antibodies in the serum samples of dogs, Tao and Li (2014) developed an immunochromatographic test strip using colloidal gold-coated staphylococcal protein A (SPA). The specificity and sensitivity of the test is 93.1% and 92.2%, respectively.…”

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

“…A method for measuring hepatitis B virus surface antigen in serum was created by using a chromatographic sandwich immunoassay with fluorescence detection [91]. A test strip utilizing this general approach was developed for the detection of rabies in canine serum [92]. Quantum dots were employed in a sandwich immunoassay format for the analysis of C-reactive protein [93], and chromatographic sandwich immunoassays have been made for detecting lily mottle virus and lily symptomless virus [94,95].…”