Development of a latex agglutination test for norovirus detection

Abstract

Norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Currently, reverse transcription polymerase chain reaction (RT-PCR) is used commonly to detect NoVs in both clinical and environmental samples. However, RT-PCR requires expensive equipment and cannot be performed on site. In this study, a latex agglutination test (LAT) using antibody-labeled latex beads for detecting NoVs was developed. Two kinds of polyclonal antibodies, one generated from synthetic peptides and the other from E. coli-expressed NoV capsid proteins, were used to develop the LAT. Each of these polyclonal antibodies was immobilized on the surface of latex beads and tested for the ability to detect NoVs. Under optimized conditions, our LAT detected GII.4 NoV at concentrations as low as 3.3x10(5) RT-PCR units/ml in stool samples. The detection limit for the LAT was approximately 1.7 103 RT-PCR units. Forty-eight stool samples were tested for NoVs using this LAT. In comparison with an RT-PCR assay, the sensitivity and specificity of the LAT were 35% and 100%, respectively. With further optimization, this LAT used with appropriate antibodies could be applied for convenient detection of NoVs in clinical diagnosis and food monitoring.

The validation and utility of a quantitative one-step multiplex RT real-time PCR targeting Rotavirus A and Norovirus

“…Viruses, specifically Rotavirus A (RoV) and genogroup I and II Norovirus (NoVI and II) are predominant causes of viral gastroenteritis worldwide, and are responsible for over 40% of all cases of diarrhea in developing countries (Davidson et al, 2002; Patel et al, 2008; Widdowson et al, 2009). A variety of techniques are used to detect RoV and NoV in stool samples, including electron microscopy (Caul, 1996a,b), latex agglutination (Lee et al, 2010), PCR amplification, and enzyme immunoassay (EIA) (Jiang et al, 2000). Commercial EIA kits are the most common methods used for diagnosis, as they offer simplicity and good specificity, but may lack overall sensitivity in some settings (Barreira et al, 2010; Ramani et al, 2010).…”

Rapid Waterborne Pathogen Detection with Mobile Electronics

“…Traditionally, immunoagglutination is only used in a qualitative test; however, together with the narrow beam technique, immunoagglutination offers a simple, generally applicable, and non-hazardous method for fast and bacterium-specific quantitative detection [33,34,35,36,37,38,39,40,41,42,43,44,45,46]. More importantly, the agglutination reaction is a one-step reaction method mediated by specific reactions between antibodies immobilized on microbeads and antigens in the sample, requiring no further washing steps prior to detection.…”

Comparison of methods for the detection of coliphages in recreational water at two California, United States beaches

“…There have been improvements introduced such as the use of different types of beads and more directional binding of the immunoglobulin (Inzana, 1995;Molina-Boívar et al, 1998;Perez-Amodio et al, 2001), which may improve performance of the method used in this study. The development of CLAT assay has spurred others to develop a latex agglutination method for Norovirus (Lee et al, 2010).…”