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 59 concentration (laboratory standard). Agreement of HPV detection was 89Á8% (j = 0Á798; P = 0Á007), with HPV detected in 10 more samples for the MP96. There was a high concordance of detection for any oncogenic HPV genotype (j = 0Á77; P = 0Á007) and for any low-risk HPV genotype (j = 0Á85; P = 0Á008). DNA extracted at laboratory standard had a lower overall agreement 85Á2% (j = 0Á708; P < 0Á001), with 17/115 discordant positive samples that tested negative after STARlet extraction. Of the discordant genotypes, 72Á7% 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.
Context.—
Detection of human papillomavirus (HPV) in formalin-fixed, paraffin-embedded (FFPE) tissues may identify the cause of lesions and has value for the development of new diagnostic assays and epidemiologic studies. Seegene Anyplex II assays are widely used for HPV screening, but their performance using FFPE samples has not been fully explored.
Objective.—
To validate Anyplex II HPV HR Detection (Anyplex II, Seegene) using FFPE samples.
Design.—
We used 248 stored DNA extracts from cervical cancer FFPE samples collected during 2005–2015 and that tested HPV positive using the RHA kit HPV SPF10-LiPA25, v1 (SPF10, Labo Biomedical Products) HPV genotyping assay, manufacturer-validated for FFPE samples.
Results.—
Of the selected 248 samples, 243 were used in our analysis. Consistent with SPF10 genotyping results, Anyplex II detected all 12 oncogenic types and had an overall HPV detection rate of 86.4% (210 of 243 samples). Anyplex II and SPF10 showed very high agreement for the detection of the 2 most important oncogenic genotypes: HPV 16 (219 of 226; 96.9%; 95% CI, 93.7–98.75) and HPV 18 (221 of 226; 97.8%; 95% CI, 94.9–99.3).
Conclusions.—
Overall results showed that both platforms produced comparable HPV genotyping results, indicating the suitability of Anyplex II for FFPE samples. The Anyplex II assay has the added convenience of being an efficient, single-well semiquantitative polymerase chain reaction assay. Further optimization of Anyplex II may enhance its performance using FFPE samples by improving the detection limit.
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