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Disclaimer: Early release articles are not considered as final versions. Any changes will be reflected in the online version in the month the article is officially released.

Volume 32, Number 10—October 2026

Research

Isothermal Detection of Influenza D Virus using Reverse Transcription Loop-Mediated Isothermal Amplification

Carlos Abelardo dos Santos, Jialu Li, Pauline M. van Diemen, Andrew McMahon, Benjamin C. Mollett, Andrew M. Ramsay, Meshach M. Maina, Joe James, Helen E. Everett, Janet M. Daly, and Nicole C. RobbComments to Author 
Author affiliation: Warwick Medical School, University of Warwick, Coventry, UK (C.A. dos Santos, J. Li, N.C. Robb); Animal and Plant Health Agency Weybridge, Addlestone, UK (P.M. van Diemen, B.C. Mollett, A.M. Ramsay, J. James, H.E. Everett); School of Life Sciences, University of Warwick, Coventry (A. McMahon); One Virology, Wolfson Centre for Global Virus Research, University of Nottingham, Nottingham, UK (M.M. Maina, J.M. Daly)

Main Article

Figure 4

Specificity and sensitivity assays for influenza D virus (IDV) detection in study of isothermal detection of IDV using reverse transcription loop-mediated isothermal amplification (RT-LAMP). A) The total of 106 copies/µL of gBlocks containing the influenza A, B, C and D virus polymer basic protein 2 gene sequences were tested. Only the gBlocks containing the target sequences were amplified, as demonstrated by color change in reactions. B) Two percent gel electrophoresis of 1 representative of each amplification products of the RT-LAMP reactions shown in panel A, showing the typical lamp ladder-like pattern in IDV virus samples and no visible nonspecific bands in the other target samples. C) Dilutions of target RNA in 8 replicates for sensitivity analysis at different concentrations, varying from 500 copies per µL (cp/μL) of target (top row) to 10 cp/μL of target (bottom row). D) Probit-analysis curve of probability of detection in relation to the number of copies. The limit of detection is defined as the minimum number of copies detected >95% of the time. NTC, nontemplate control.

Figure 4. Specificity and sensitivity assays for influenza D virus (IDV) detection in study of isothermal detection of IDV using reverse transcription loop-mediated isothermal amplification (RT-LAMP). A) The total of 106 copies/µL of gBlocks containing the influenza A, B, C and D virus polymer basic protein 2 gene sequences were tested. Only the gBlocks containing the target sequences were amplified, as demonstrated by color change in reactions. B) Two percent gel electrophoresis of 1 representative of each amplification products of the RT-LAMP reactions shown in panel A, showing the typical lamp ladder-like pattern in IDV virus samples and no visible nonspecific bands in the other target samples. C) Dilutions of target RNA in 8 replicates for sensitivity analysis at different concentrations, varying from 500 copies per µL (cp/μL) of target (top row) to 10 cp/μL of target (bottom row). D) Probit-analysis curve of probability of detection in relation to the number of copies. The limit of detection is defined as the minimum number of copies detected >95% of the time. NTC, nontemplate control.

Main Article

Page created: August 13, 2026
Page updated: September 21, 2026
Page reviewed: September 21, 2026
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