An isothermal, multiplex amplification assay for detection and genotyping of human papillomaviruses in formalin-fixed, paraffin-embedded tissues Journal Article

   

Authors:	Tang, Y. W.; Lozano, L.; Chen, X.; Querec, T. D.; Katabi, N.; Moreno-Docón, A.; Wang, H.; Fix, D.; De Brot, L.; McMillen, T. A.; Yoon, J. Y.; Torroba, A.; Wang, Y.; Unger, E. R.; Park, K. J.
Article Title:	An isothermal, multiplex amplification assay for detection and genotyping of human papillomaviruses in formalin-fixed, paraffin-embedded tissues
Abstract:	Rapid and accurate identification of human papillomavirus (HPV) is important for both clinical management and population screening. Analytic validation of Atila AmpFire Multiplex HPV assays on formalin-fixed, paraffin-embedded (FFPE) cervix/vulva and oropharynx diagnostic tissue samples was performed. The AmpFire assay incorporates a novel isothermal multiplex amplification coupled with real-time fluorescent detection to detect and genotype 15 high-risk (HR) HPV genotypes. Limits of detection determined by plasmids cloned with HPV genotype-specific sequences were 2 copies/reaction for HPV16, HPV18, and some HR HPV genotypes, and 20 copies/reaction for the remaining HR HPV genotypes. The performance of the AmpFire assays in clinical samples was evaluated using 214 FFPE specimens. The AmpFire assay failed in one clinical specimen for an invalid rate of 0.5%. The AmpFire assay detected HPV in clinical samples with positive percent agreements of 100.0% for HPV16, 100.0% for HPV18, and 94.7% for non-16/18 HR HPV, and 100% negative percent agreements for HPV16, HPV18, and non-16/18 HR HPV. Qualitative detection agreement was obtained in the reproducibility study. In summary, the Atila AmpFire HPV assay demonstrated excellent analytic sensitivity and specificity for detection and genotyping of 15 HR HPV genotypes. Assay parameters of simple specimen processing, small sample size requirement, rapid turnaround time, and being near instrument-free render it well suited for HPV detection and genotyping in FFPE specimens. © 2020 American Society for Investigative Pathology and the Association for Molecular Pathology
Keywords:	controlled study; gene sequence; human cell; nonhuman; diagnostic accuracy; genotype; mass screening; molecular cloning; real time polymerase chain reaction; dna extraction; virus detection; wart virus; molecular pathology; limit of detection; human papillomavirus type 16; human papillomavirus type 18; human; female; article; hela cell line; human papillomavirus type 33; ca ski cell line; human papillomavirus type 31; human papillomavirus type 35; human papillomavirus type 39; human papillomavirus type 45; human papillomavirus type 51; human papillomavirus type 52; human papillomavirus type 53; human papillomavirus type 56; human papillomavirus type 58; human papillomavirus type 59; human papillomavirus type 66; human papillomavirus type 68
Journal Title:	Journal of Molecular Diagnostics
Volume:	22
Issue:	3
ISSN:	1525-1578
Publisher:	Elsevier Science, Inc.
Date Published:	2020-03-01
Start Page:	419
End Page:	428
Language:	English
DOI:	10.1016/j.jmoldx.2019.12.004
PUBMED:	31978559
PROVIDER:	scopus
PMCID:	PMC7081142
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-85080893255&doi=10.1016%2fj.jmoldx.2019.12.004&partnerID=40&md5=5f3991351e00599e379b98f6346c2e1f
Notes:	Article -- Export Date: 1 April 2020 -- Source: Scopus

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