Development of RT-qPCR for quantification of human enterovirus D68 in vitro

The common cold is the most frequent viral infectious disease of the upper respiratory tract with different intensities based on the serotype and the characteristics of the virus. Numerous human rhinoviruses have been identified and classified. Human rhinovirus 87 (HRV87), also known as enterovirus D68 (EV-D68), is one of the common viruses causing respiratory infections. In this study, a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay was developed, optimized, and validated for the detection of EV-D68. Method development also covers specificity, sensitivity, efficiency, and inter-and-intra-assay variations. Overall, this one-step qPCR assay will permit quantitative assessments of human enterovirus D68 RNA. • Enterovirus D68 is a reemerging viral agent causing respiratory infection. • RT-qPCR assay developed for detection of human enterovirus D68. • In this article validation to secure reproducibility is done according to MIQE guidelines.