Ngoc Yen Kim Tran1*, Thuy Phan1, Genevieve McKew1,2 and Steven Siarakas1
1Department of Microbiology and Infectious Disease, Concord Repatriation General Hospital, NSW Health Pathology, Concord, NSW, Australia 2Faculty of Medicine and Health, The University of Sydney, Concord, NSW, Australia
1Department of Microbiology and Infectious Disease, Concord Repatriation General Hospital, NSW Health Pathology, Concord, NSW, Australia
2Faculty of Medicine and Health, The University of Sydney, Concord, NSW, Australia
*Corresponding Author: Ngoc Yen Kim Tran, Department of Microbiology and Infectious Disease, Concord Repatriation General Hospital, NSW Health Pathology, Concord, NSW, Australia.
Received: February 17, 2022; Published: March 29, 2022
A validation study was conducted on the AusDiagnostics MT-Prep XL and AusDiagnostics SARS-CoV-2, Influenza and RSV 8-well assay performed on the AusDiagnostics Ultraplex 3 system. The Cycle threshold (Ct) of clinical patient samples spiked with a positive control, was compared to the reference laboratory standard, AusDiagnostics MT-Prep and Highplex System. The analytical performance was evaluated to establish the limit of detection (LOD), precision and agreement.
There is an average difference in Ct of -0.883 and 0.078 between the nucleic extractors and MT-PCR systems, respectively. There was strong correlation (R-value ranging from 0.78 – 0.91 for all gene targets) between the AusDiagnostics Ultraplex-3 and Highplex. Bland Altman analysis demonstrated the overall differences between the assays are within the limits of agreement (LoA). The AusDiagnostics MT-Prep XL has a processing time of 1.04 minute/sample and AusDiagnostics Ultraplex 3, 1.93 minute/sample, demonstrating a higher efficiency rate compared to the reference standard.
RSV and SARS-CoV-2 detection in clinical samples were compared to the AusDiagnostics Highplex system as a reference standard. The assay demonstrated 99.10% overall agreement, with a PPA 100% and NPA of 99%. Kappa statistics of 0.95 (p < 0.05) suggests excellent agreement level between the two platforms. Moreover, precision study was conducted to observe the intra-run and inter-run assay variability. The calculated average Ct observed in all targets is 1.063% and 1.682%, respectively. The CV% is <2% suggesting high precision, low variability, and dispersion in data points.
The commercialized AusDiagnostics MT-Prep XL and Ultraplex 3 is a viable platform, fit for purpose in detecting the SARS-CoV-2, Influenza A, Influenza B and RSV respiratory viruses.
It’s high efficiency rate and 96-sample capacity enhances laboratory throughput and enables faster time to result in comparison to the current reference standard.
Keywords: COVID-19; Influenza; Respiratory Syncytial Virus; Laboratory Diagnosis; MT-PCR
Citation: Ngoc Yen Kim Tran., et al. “Evaluation and Validation of the AusDiagnostics MT-Prep XL and AusDiagnostics Ultraplex-3 [SARS-CoV-2, Influenza and RSV 8-well Assay]". Acta Scientific Microbiology 5.4 (2022): 156-165.
Copyright: © 2022 Ngoc Yen Kim Tran., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.