Development of a simple and rapid latex test for rotavirus in stool samples

Abstract

In the fields of infectious diseases, a variety of immunoassay methods has been reported. More simple, rapid and cost-effective manual tests for infectious diseases are needed. For the purpose of simple diagnosis of rotavirus infection, a rapid test against human rotavirus was developed. As for the preparation of the reagent, clinically isolated human rotavirus was cultured using MA-104 cells. Then, the cultured virus was purified by an ultracentrifugation method. The purified virus was inactivated with formaldehyde for use as antigen for immunization of rabbits. The antibody was purified and applied to sensitize the surface of latex particles to react with rotavirus in diarrheal stools. The latex agglutination test showed that the sensitivity was 98.2% and the specificity was 94.8%, in comparison with electron microscopy as the gold standard. Furthermore, to measure the virus concentration in stool samples, an automated latex photometric immunoassay system (LPIA) was used, which has been developed for quantitative measurement of agglutination. The range of the reactive viruses detected by the LPIA system was 3 x 10(7)-10(9) virions/mL. The rotavirus-positive and rotavirus-negative subjects were clearly discriminated. The results were in good accordance with electron microscope results. Those results showed that the latex agglutination test is a good tool for the simple and rapid detection of rotavirus in stool samples.

Investigation of avian rotavirus infections in broiler chicks from commercial flocks with different performance efficiency indexes

“…The lowest concentration of rotavirus detected in human fecal samples by ss-PAGE was 10 8 virions/mL (Kohno et al 2000 ). Due to the low sensitivity of the ss-PAGE technique (Otto et al 2006 ), positive results obtained in this assay indicate a high viral excretion at the time of sampling.…”

Detection of Anti-GAD65 Antibodies in Sera of Diabetic Patients Using a Home-Made Latex Agglutination Kit

“…), pH was optimized at 7, and the mixture was incubated at 37 0 C for 20 hours. After that, the mixture was ready for use as latex agglutination test reagent for the detection of anti-GAD 65 antibodies and stored at 4 0 C. [(Hideki et al 2000), with some modifications] (12) .…”

Development of an enzyme-linked immunospot assay for determination of rotavirus infectivity

“…Detection of rotaviruses depends on electron microscopy (Brandt et al, 1981), enzyme-linked immunosorbant assay (Beards et al, 1984), latex agglutination (Kohno et al, 2000), lateral-flow immunoassay (Al-Yousif et al, 2002) and nucleic acid amplification (RT-PCR) (Kang et al, 2004;Ranheim et al, 2006). However, cell culture method remains the gold standard for virus diagnosis because it is the only method for detection of infectious viral particles.…”