ABSTRACT Background Rotavirus (RV) is the leading cause of severe gastroenteritis in children and is responsible for significant morbidity and mortality in developing and low‐income countries. Hence, precise and prompt diagnosis of rotavirus infection is vital for treatment. The currently employed detection methods include rotavirus real‐time reverse transcriptase polymerase chain reaction (RT‐qPCR) and antigen‐capture enzyme‐linked immunosorbent assay (Ag‐ELISA) which have some known strengths and weaknesses. Methods Rotaviruses are difficult to grow in various cell substrates; however, a few cell lines, including MA104, show evident virus growth. Hence, MA104 cell‐based immunocolorimetric assay (ICA) was developed, and its sensitivity and specificity were studied with the existing rotavirus detection methods that target the viral antigen or nucleic acid. A total of 84 fecal specimens obtained from acute gastroenteritis children were utilized, and the results of ICA were compared with commercial Ag‐ELISA and in‐house RT‐qPCR. Results A cell culture‐based RV‐ICA was successfully standardized and validated with commercial Ag‐ELISA and in‐house RT‐qPCR. The sensitivity and specificity of ICA with commercial Ag‐ELISA were found to be 90.7% and 36.6%, respectively. The comparison of RT‐qPCR with RV‐ICA showed slightly lower sensitivity, that is, 84.3%, whereas increased specificity, that is, 57.1%. The positive predictive value of RV‐ICA with Ag‐ELISA and RT‐qPCR was 60% and 90.7%, respectively. It is well accepted that RT‐qPCR is a more sensitive and rapid assay for virus detection due to amplification and quantification of target nucleic acid. However, a cell culture‐based ICA has an added advantage over Ag‐ELISA and RT‐qPCR, due to added possibility of rotavirus isolation from positive fecal specimens. Conclusions The standardized cell culture‐based RV‐ICA supports the detection of live rotaviruses from fecal specimens over the commercial Ag‐ELISA and RT‐qPCR, thus helps identify samples suitable for rotavirus isolation.
Kumbhar et al. (Sat,) studied this question.