Evaluation of the Roche MagNA Pure 96 nucleic acid extraction platform for the Seegene Anyplex II HPV28 detection assay

Atchison, S. and Shilling, H. and Balgovind, P. and Machalek, D. A. and Hawkes, D. and Garland, S. M. and Saville, M. and Murray, G. and Molano, M. and Danielewski, J. and Phillips, S. (2021) Evaluation of the Roche MagNA Pure 96 nucleic acid extraction platform for the Seegene Anyplex II HPV28 detection assay. Journal of Applied Microbiology, 131 (5). pp. 2592-2599. ISSN 1364-5072, DOI https://doi.org/10.1111/jam.15126.

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Aim Validate the Roche, MagNAPure96 (MP96) nucleic acid extraction platform for Seegene Anyplex II HPV28 (Anyplex28) detection of Human Papillomavirus. Methods and Results Comparisons were made for Anyplex28 genotyping from 115 cervical samples extracted on the Hamilton, STARlet and the MP96. Two DNA concentrations were used for the MP96, one matched for sample input to the STARlet and another 5x concentration (laboratory standard). Agreement of HPV detection was 89 center dot 8% (kappa = 0 center dot 798; P = 0 center dot 007), with HPV detected in 10 more samples for the MP96. There was a high concordance of detection for any oncogenic HPV genotype (kappa = 0 center dot 77; P = 0 center dot 007) and for any low-risk HPV genotype (kappa = 0 center dot 85; P = 0 center dot 008). DNA extracted at laboratory standard had a lower overall agreement 85 center dot 2% (kappa = 0 center dot 708; P < 0 center dot 001), with 17/115 discordant positive samples that tested negative after STARlet extraction. Of the discordant genotypes, 72 center dot 7% were detected in the lowest signal range for Anyplex28 ('+'). Conclusions MP96 performed with high concordance to STARlet, although produced DNA with a higher analytical sensitivity on the Anyplex28. Significance and Impact of the Study This analysis supports the use of samples extracted on the MP96 for HPV genotyping using the Anyplex28. Furthermore, an increase in DNA concentration increased analytical sensitivity of the Anyplex28, particularly appropriate for prevalence studies.

Item Type: Article
Funders: Commonwealth Department of Health HPV Surveillance Fund[4-8FG4IWD]
Uncontrolled Keywords: Detection;Diagnosis;Diseases;Molecular genetic;Polymerase chain reaction
Subjects: R Medicine
Depositing User: Ms Zaharah Ramly
Date Deposited: 15 Jun 2022 04:13
Last Modified: 15 Jun 2022 04:13
URI: http://eprints.um.edu.my/id/eprint/34420

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