WHO/BS/2016.2299 Collaborative Study to Establish a WHO International Reference Panel for Dengue Virus types 1 to 4 RNA for Nucleic Acid Amplification Technology (NAT)-Based Assays

Overview

An international collaborative study was conducted to assess the suitability of a candidate reference panel for Dengue virus (DENV) types 1 to 4 RNA for use in nucleic acid amplification technology (NAT)-based assays. Two reference panel candidates were prepared for each DENV type, one liquid frozen (Panel 1) and one lyophilized (Panel 2). The reference panel candidates consisted of a DENV prototype laboratory strain that was grown in cell culture in mosquito cells, subjected to inactivation of infectivity by heat treatment and diluted in human plasma. Both reference panels were sent coded to the participants for testing in four independent runs utilizing the DENV NAT assay available in their laboratory. Results were communicated to the U.S. Food and Drug Administration (FDA) where the analyses were performed. A total of 28 laboratories from 20 countries accepted to participate in the study, of which 21 submitted the results for qualitative and quantitative assessments.

It is proposed that Panel 1, which consists of heat-inactivated and lyophilized preparations of cell culture-derived DENV-1(Hawaii, GenBank# KM204119), DENV-2 (New Guinea C, GenBank # KM204118), DENV-3 (H87, GenBank# KU050695 ), and DENV4 (H241 GenBank# KR011349 ) strains be established as the 1 st International Reference Panel for DENV RNA with a unitage of 13,500, 69,200, 23,400, and 33,900 International Units per ml for DENV-1 to 4, respectively. The real-time and accelerated stability studies of the proposed International Reference Panel revealed that the material is stable at the recommended storage temperature, i.e. at or below 4°C, for 12 months, and is therefore suitable for long term use.