Development of a reverse transcription loop-mediated isothermal amplification assay (RT-LAMP) that detects enteroviruses by targeting the highly conserved 5 ′-UTR region

In this study, we developed a rapid, simple, sensitive, and specific isothermal reverse transcription assay (RT-Loop -Mediated Amplification, RT-LAMP) for the detection ofEV-A, B, C, and D species of enteroviruses, by targeting the highly conserved 5 ′UTR region. The assay was designed and validated based on reference sequences of the four species and on clinical and environmental isolates. The limit of detection of the assay is 0.75 CCID50/assay forEnterovirus A, B, andD and 0.075 CCID50/assay forEnterovirus C. LAMP allows immediate diagnosis in just 30 –50 min, instead of a minimum of 120 min needed for PCR with an equal or better sensitivity. Moreover, due to its isothermal nature, there is no need for expensive equipment, thus decreasing the cost of each reaction. Therefore, this assay is ideal for use in resource-limited settings such as pr imary care facilities and environmental and clinical laboratories in developing countries.
Source: Virus Genes - Category: Genetics & Stem Cells Source Type: research