Comparison of the AmpFire® Multiplex HPV Assay to the Xpert® HPV Assay for detection of human papillomavirus and cervical disease in women with human immunodeficiency virus: A pragmatic performance evaluation

Abstract

Background: Low- and middle-income countries (LMICs) account for nearly 85% of the global cervical cancer burden, yet have the least access to high-performance screening. International guidelines recommend human papillomavirus testing (HPV) as primary screening, yet implementation is inhibited by the cost of HPV testing.Atila AmpFire® HPV Assay (AmpFire) is both affordable and easy to use, and offers individual genotyping. The objective of this study was to compare the performance of the AmpFire HPV assay to the Xpert® HPV assay in detection of both HPV and clinically significant cervical disease. Methods: We utilized stored cervical specimens from a prospective cohortstudy of women living with human immunodeficiency virus (HIV) in Botswana conducted from May to July 2018. Positive and negative percent agreement was calculated for the AmpFire and Xpert assays, as was detection of high-grade cervical dysplasia. Results: 63 stored cervical specimens haddetectable DNA after thawing and were included in the analysis. The positive percent agreement was 91.2% (95%CI: 76.3-98.1) and negative percent agreement was 79.3% (95% CI: 60.3-92.0). Six cases positive by AmpFire but negative by Xpert were HPV genotypes 35, 52 (n=2), 58, 68, and co-infection with HPV 45 and 68. Both Xpert and AmpFire assays detected HPV in all 10 samples of women who had high-grade cervical dysplasia. Conclusions: The AmpFire HPV assay demonstrated excellent analytic performance in both detection of HPV and clinically significant cervical disease. AmpFire HPV is a promising option to increase access to affordable, type-specific HPV screening for cervical cancer in LMICs.

Intra‐ and interlaboratory reproducibility evaluation toward international validation status of the AmpFire assay

“…In Moyo et al 23 data from the “AmpFire assay vs Xpert® assay” comparison in histopathology samples of a Botswanan population ( n = 63) were found. Because of the small sample size and a bias in using histopathological samples, data were not extracted.…”

“…In the case of CIN3+, the sensitivity was 100.0% (95% CI: 88.3%-100.0%). The relative sensitivity for CIN2+ of the AmpFire assay versus the cobas 4800 HPV assay was 1.034 In Moyo et al 23 data from the "AmpFire assay vs Xpert ® assay" comparison in histopathology samples of a Botswanan population (n = 63) were found. Because of the small sample size and a bias in using histopathological samples, data were not extracted.…”

“…Based on the literature review, the AmpFire assay was mentioned in six publications. [22][23][24][25][26][27] In the Chinese multicentre screening trial (CHIMUST) study-containing 6042 Chinese women (including 57 CIN2 and 35 CIN3 cases) with a mean age of 44.1 years old-that assessed the clinical accuracy of a previous version of the AmpFire assay (including HPV53) 22 using cobas 4800 HPV assay (Roche Molecular Diagnostics) as comparator, which was recently recognized as a valid HPV comparator. 16 In the CHIMUST study, a statistical correction was applied by considering removal of all women having a single HPV53 from the analysis.…”