Obtaining of a rapid diagnostic test for Cholera, based on latex particles coupled with a monoclonal antibody against Vibrio cholera O1 lipopolysaccharide

Abstract

Cholera is an acute contagious intestinal disease caused by ingestion of food or water contaminated with O1 and O139 serotypes of the bacterium Vibrio cholerae. Cholera is characterized by abundant secretory diarrhea leading to dehydration. Death occurs within hours without treatment, so early diagnosis is very important, especially at the beginning of the disease, because it is difficult to differentiate from other acute diarrheal diseases. The diagnostic golden test is the stool culture; however, it does not guarantee a rapid detection of the disease. Rapid tests have been recently developed; they are based on test strips and agglutination with latex particles, which are very effective, but difficult to acquire for their high prices. The objective of this research was to obtain a quick assay based on latex particles coupled with a monoclonal antibody (mAb) against V. cholerae O1 lipopolysaccharide obtained in Finlay Institute. Latex particles of 0.8 µm were used in a 10% suspension, and they were coupled to the mAb (0.25 mg/ml) for 2 hours at 37°C. The sensitivity, specificity and performance were evaluated in 84 stool samples from patients with presumptive diagnosis of cholera. The diagnostic test obtained showed no cross-reactivity against no-O1 strains and other enteropathogens. Latex diagnostic test showed values of sensitivity, specificity and efficacy of 97.87; 97.29 and 97.6% respectively, very similar to the commercial diagnostic test CTK- Biotech. The latex reagent obtained can be used in the rapid diagnosis of the disease.

Keywords

• monoclonal antibodies a
• Vibrio cholerae O1 a
• diagnostic a
• latex