Evaluation of a loop-mediated isothermal amplification method for rapid detection of human respiratory syncytial virus in children with acute respiratory infection

Abstract

Introduction: Human Respiratory Syncytial Virus (hRSV) is the most frequent cause of acute respiratory infection of the lower respiratory tract in children under five years of age. The development of molecular techniques to identify hRSV is one of the current challenges in the field of clinical research. Objective: To evaluate an isothermal amplification method for the rapid detection of respiratory syncytial hVSR in children with acute respiratory infection. Materials and methods: Viral RNA was extracted from 304 samples by nasal swab (NP) in children with symptoms of acute respiratory infection who attended the emergency service of the University Hospital of the North in the city of Barranquilla during the months of April 2016 to July 2017. The Reverse Transcriptase-LAMP assay (RT-LAMP (M)) was evaluated against molecular techniques such as polymerase chain reaction-multiple nested reverse transcriptase (Nested RT- PCR), which the National Institute of Health (NIH) Colombia routinely applied and was used as a standard; quantitative PCR (qPCR) and Reverse Transcriptase-LAMP (RT-LAMP (L)) for the rapid response of hRSVA and hRSVB. Results: The RT-LAMP (M) assay had a sensitivity (93.59%), a specificity (92.92%) and a concordance of (0.83 ± 0.036) with the nested RT-PCR test. The Kappa index of RT-LAMP (M) was higher, while the RT-LAMP (L) and qPCR values are in agreement (0.75 ± 0.043 and 0.71 ± 0.045, respectively). Conclusion: These results indicate that RT-LAMP (M) can be considered as a tool of clinical utility to detect hRSVA, considering the shorter time it takes to obtain results, lower costs and better performance than the other molecular tests evaluated.