Comparison of Luminex NxTAG Respiratory Pathogen Panel and xTAG Respiratory Viral Panel FAST Version 2 for the Detection of Respiratory Viruses

Lee, Chun Kiat; Lee, Hong Kai; Ng, Christopher Wei Siong; Chiu, Lily; Tang, Julian Wei-Tze; Loh, Tze Ping; Koay, Evelyn Siew-Chuan

All samples were initially tested with the xTAG RVP FAST v2 as part of our routine clinical service. In brief, the extracted nucleic acid (10 µL) was used for target amplification by multiplex reverse transcription PCR (RT-PCR). The PCR product (2 µL) was hybridized to a bead mix; next, reporter dye was added in a new reaction vessel, which was sealed and incubated. The amplification and hybridization/incubation were performed on the Applied Biosystems Veriti thermal cycler (Thermo Fisher Scientific, Wohlen, Switzerland), as per the manufacturer's recommendations. Signal acquisition was performed on the MAGPIX instrument (Luminex Corp). After testing, the extracted nucleic acids were immediately frozen at -80°C until further testing. Residual frozen archival samples were retrieved and tested with the NxTAG RPP, a closed-tube nucleic acid assay containing premixed lyophilized reagents for target amplification, PCR product hybridization/incubation, and detection. All procedures were carried out according to the manufacturer's instructions. The extracted nucleic acid (35 µL) was added to resuspend the preplated lyophilized bead reagents in the vessel. Multiplex RT-PCR, bead hybridization, and reporter dye incubation were performed on the Veriti thermal cycler, as per the manufacturer's recommendations. Finally, the vessel was placed onto the MAG-PIX instrument for signal acquisition.