title: Rapid detection of human rotavirus using NSP4 gene specific reverse transcription loop-mediated isothermal amplification assay.
abstract: The seasonal outbreaks of human rotavirus (RV) infection occur every winter. Most patients are diagnosed clinically by a rapid latex agglutination detection kit or polymerase chain reaction assays for RV from stool samples, but some problems have been reported on the specificity and sensitivity of such rapid detection assays. To ratify these issues, a sensitive, specific, simple, and rapid nucleic acid based diagnostic method is expected to be introduced and the reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect the RV in human stool samples by incubation at 60 °C for 1 h and amplification was confirmed by electrophoretic laddering, restriction enzyme digestion, and hydroxynapthol blue discoloration. The assay established in this study was found to detect only the RVs and no cross-reaction with other viruses, demonstrating its high specificity. By using serial samples dilution as template, the detection limit of LAMP was 10 times more than that of PCR. The results showed the potential clinical feasibility of RT-LAMP as a useful diagnostic tool for the detection of RV with high sensitivity in comparison to conventional RT-PCR.
authors.affiliation: Indian Veterinary Research Institute, Bareilly, Uttar Pradesh India.
authors.forename: Yashpal Singh
authors.initials: YS
authors.lastname: Malik
authors.affiliation: Indian Veterinary Research Institute, Bareilly, Uttar Pradesh India.
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Sharma
authors.affiliation: Indian Veterinary Research Institute, Bareilly, Uttar Pradesh India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Indian Veterinary Research Institute, Bareilly, Uttar Pradesh India.
authors.forename: Sathish B
authors.initials: SB
authors.lastname: Shivachandra
authors.affiliation: Government Medical College, Haldwani, Uttarakhand India.
authors.forename: Vinita
authors.initials: V
authors.lastname: Rawat
authors.affiliation: Government Medical College, Haldwani, Uttarakhand India.
authors.forename: Ritu
authors.initials: R
authors.lastname: Rakholia
authors.affiliation: Project Directorate on Foot and Mouth Disease, Mukteswar, Nainital, 263138 Uttarakhand India.
authors.forename: Rajeev
authors.initials: R
authors.lastname: Ranjan
authors.affiliation: Division of Virology, National Institute of Cholera & Enteric Diseases (NICED), Indian Council of Medical Research (ICMR), P-33, C.I.T. Road, Scheme XM, Beliaghata, Kolkata, 700010 India.
authors.forename: Balasubramanian
authors.initials: B
authors.lastname: Ganesh
authors.affiliation: Defence R & D Establishment DRDE, DRDO, Ministry of Defence, Jhansi Road, Gwalior, 474002 Madhya Pradesh India.
authors.forename: Manmohan
authors.initials: M
authors.lastname: Parida
keywords: Group A; Human; NSP4 gene; RT-LAMP; Rotavirus