Ngoc Yen Kim Tran¹*, Thuy Phan¹, Genevieve McKew^1,2 and Steven Siarakas¹

¹Department of Microbiology and Infectious Disease, Concord Repatriation General Hospital, NSW Health Pathology, 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

Abstract

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

References

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Citation

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

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.