Comparative evaluation of antigen ELISA technique and PCR for detection of Rotavirus in stool samples of pediatric patients with diarrhea

Background: Acute diarrheal disease is a common infection of young children and adults which is characterized by rapid dehydration of body fluid. Iidentification of the causative agent and timely treatment can be life saving of children. The objective of this study was to detect Rotavirus (RV) in stool samples and to compare ELISA and RT-PCR techniques. Methods: Hospitalised children less than 5 years admitted for acute diarrhea were examined and clinical profile of diarrhea in the children. Detection of Rotaviral antigen in stool samples was done by a commercial Enzyme Immuno Solvent Assay Premier Rotaclone kit (Meridian Bioscience, Inc.) and the samples were processed for extraction of nucleic acid using an automated extraction system and RT-PCR was performed. Results: A total of 135 stool samples were tested by ELISA and RT-PCR. A total of 20/135 (14.81%) samples were found to be positive for RV infection by ELISA. Positivity of RV by RT-PCR was 33/135 (24.44%). The positivity by ELISA was significantly lower than by RT-PCR. The sensitivity and specificity of PCR for detection of RV was 100%. Conclusion: RT-PCR is a better and sensitive technique as compared to ELISA for detection of Rotavirus. Introduction Acute diarrheal disease is a common infection of young children and adults which is characterized by rapid dehydration of body and one of the most important worldwide causes of morbidity and mortality, accounting for an estimated 1.3 million deaths in children under five aged (1,2) . Clinically, rotavirus gastroenteritis is characterized by profuse diarrhea, mild fever, and vomiting, leading to mild to severe dehydration. Rotavirus is the major cause of acute gastroenteritis and severe dehydrating diarrhea in young children (3) . Rotavirus is estimated to cause about 40% of all hospital admissions due to diarrhea among children under five years of age worldwide (4) In India rotavirus is responsible for 40% of children diarrhea hospitalization and approximately 527,000 (475,000 580,000) deaths annually (5) or 29 to 45% of all death due to diarrhea among children ≤5 year of age globally (6,7) . Several techniques have been developed to detect rotavirus in stool samples, including electron microscopy, polyacrylamide gel electrophoresis (PAGE) of viral nucleic acid, various immuno assays, and PCR-based molecular methods (1) . Several rapid methods, such as the www.jmscr.igmpublication.org Impact Factor (SJIF): 6.379 Index Copernicus Value: 71.58 ISSN (e)-2347-176x ISSN (p) 2455-0450 DOI: https://dx.doi.org/10.18535/jmscr/v6i6.143