Development of a lateral flow immunoassay strip for rapid detection of rotavirus in fecal samples

Rotavirus is the most common cause of severe, dehydrating diarrhea in children worldwide. Without treatment rotavirus infection may result in severe illness with dehydration and disturbances of the body’s normal electrolyte balance, especially in babies and preschool children. Rotavirus is the cause of up to 50% of the hospitalized cases of diarrheal illness in infants and young children and a major cause of infant mortality in the developing countries. Rapid detection of rotavirus infection could help in effective treatment of diarrheal patients. In this paper, we reported the results of the development of a lateral flow immunoassay (LFA) strip for rapid detection of rotavirus. Rabbit and guinea pig anti-rotavirus IgGs were successfully purified from serum by protein A affinity chromatography. For making the conjugate pad, optimal amount of rabbit anti-rotavirus antibody was 0.25 µg/pad. The optimal amount of guinea pig anti-rotavirus antibody for immobilization on nitrocellulose membrane was 0.1 µg at the test line. The cellulose membrane could be processed at 4 o C and 25 o C overnight or 42 o C for 30 mins after antibody immobilization. The detection limit of the generated LFA strip was 1.6 ´ 10 4 virus particles. The produced LFA strips were tested with 30 fecal specimens and compared with ELISA assay.