A novel real-time PCR assay panel for detection of common respiratory pathogens in a convenient, strip-tube array format

Mohammad Rubayet Hasan; Hassan Al Mana; Virginia Young; Patrick Tang; Eva Thomas; Rusung Tan; Peter Tilley

Respanel PCR results in 91%-100% agreement with the reference results (Supplemental table 3) . 223 Next, a total of 34 nasopharyngeal wash specimens that were previously tested by 224 Resplex I and II were tested by Respanel assay as described in the materials and methods. The 225 overall, observed agreement with Resplex assay was ~95% (kappa = 0.734; 95%CI: 0.643-226 0.826). In total, 71 pathogens were identified in these samples by Respanel, which was 48% 227 higher than that of Resplex assays ( for each of the individual pathogens were compared, Respanel assay results were more correlated 240 (>95%) to FTDRP21 assay than Resplex assays (Table 1 and 2).The cost of Respanel is 241 approximately 25% of Resplex assays and requires less than 10 minutes of hands-on time per 242 sample, apart from the initial effort required for producing PCR-ready aliquots of primer/probe 243 mixes in PCR strip-tubes. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/455568 doi: bioRxiv preprint of automation. Many test methods have also been described in the literature for potential use as 254 laboratory-developed tests (LDT). To make an appropriate choice, a careful assessment of 255 performance characteristics as well as costs and benefits associated with the test method is 256 necessary. Ideally, when performance characteristics of test methods are comparable and meet 257 regulatory requirements, a test method and a platform that provides rapid TAT and requires 258 minimum technical skill and hands on time would be preferable. However, the costs associated 259 with the platform, such as test kits and service contracts may not be affordable to many The purpose of this study was to develop a laboratory-developed, real-time PCR based 271 test panel for selected respiratory pathogens with minimum multiplexing and in a format so that 272 minimum hands-on time is required. Accordingly, we designed our workflow so that assay 273 specific reagents are ready to be used for routine testing (Figure 1 ). We also minimized repeated The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/455568 doi: bioRxiv preprint The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/455568 doi: bioRxiv preprint