Evaluation of a commercial immunochromatographic strip assay (ICT) for rapid detection of bovine, porcine and human Rotavirus A

The most frequent diarrhoeal pathogen “Rotavirus” is commonly detected by Ribo Nucleic Acid- Polyacrylamide Gel Electrophoresis (RNA-PAGE), Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Immunoassay (Enzyme Linked Immunosorbent Assay) in humans. However, use of these tests is limited in animals therefore, penside test like Immunochromatographic test/Lateral Flow Assay (ICT/LFA) is necessary. The present study was carried out to compare the diagnostic efficacy of rapid commercial ICT strip assay, RNA-PAGE and RT-PCR for specific detection of Rotavirus A (RVA) from stool samples of calves, piglets and children. A total of 313 faecal samples were collected between November 2022 to April 2023 from children below 5years of age (n = 100), calves (n = 100) and piglets (n = 113) ≤ 3 months of age and were screened by RNA-PAGE, RT-PCR and commercial ICT kit for rotavirus A. The overall positivity of RVA from human, calves and piglets using RNA-PAGE, RT-PCR and LFA was found to be 35 % (35/100), 8 % (8/100) and 14.16 % (16/113), respectively. Higher positivity of rotavirus was recorded in male children (44.26 %, 27/61) than in female children (20.51 %, 8/39). The kappa agreement between LFA and RT-PCR was 0.79 (substantial agreement); between LFA and RNA-PAGE was 0.61 (substantially low agreement) and between RNA-PAGE and RT-PCR was found to be 0.75 (substantial agreement). Relative diagnostic sensitivity and specificity of LFA in comparison with RT-PCR was 74.54 % and 98.44 %, respectively and in comparison with RNA-PAGE was 72.97 % and 93.47 %, respectively. RNA-PAGE in comparison with RT-PCR is having diagnostic sensitivity and specificity of 65.45 % and 99.6 %, respectively. In conclusion, the ICT strip assay is a rapid, convenient and effective method with satisfactory efficacy for detection of RVA from children, calves and piglets. ICT fulfilled the WHO ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free and Delivered) criteria for point-of-care testing. It can be useful in determining rotavirus outbreaks in resource-limited settings. • Overall RVA positivity of stool samples from human, calves and piglets using RNA-PAGE, RT-PCR and LFA was found to be 35% , 8% and 14.16, respectively. • Relative sensitivity and specificity of LFA in comparison with RT-PCR was 74.54% and 98.44% and with RNA-PAGE was 72.97% and 93.47%, respectively. • ICT/LFA is a rapid, convenient and effective method with satisfactory efficacy for detection of RVA. • ICT can be useful in determining RVA outbreaks in resource-limited settings.