A rapid HPV typing assay to support global cervical cancer screening and risk‐based management: A cross‐sectional study

Abstract

The World Health Organization recommends human papillomavirus (HPV) testing for cervical screening. Extended genotyping can identify the highest‐risk HPV‐positive women. An inexpensive, rapid, mobile isothermal amplification assay (ScreenFire HPV RS test) was recently redesigned to yield four channels ordered by cancer risk (ie, hierarchical approach): HPV16, HPV18/45, HPV31/33/35/52/58 and HPV39/51/56/59/68. Stored specimens from 2076 women (mean age 30.9) enrolled in a colposcopy clinic, with high HPV prevalence, were tested with ScreenFire. We calculated hierarchical channel positivity and non‐hierarchical channel and type positivity, according to histologic diagnosis (256 cancer, 350 cervical intraepithelial neoplasia [CIN]3, 409 CIN2, 1020 < CIN2) and known virologic reference results (Linear Array and TypeSeq). Additionally, we analyzed ScreenFire time‐to‐positive up to 60 min by channel and histology. Overall clinical sensitivity for CIN3+ was 94.7% (95% confidence interval 92.6‐96.4), similar to Linear Array (92.3, 89.7‐94.3) and TypeSeq (96.0, 93.9‐97.6). Sensitivity was high for all types and channels. The hierarchical approach was well in line with HPV typing and histologic diagnosis, prioritizing higher risk women having HPV16 and precancer. For HPV16, time‐to‐positive was shorter in women with precancer. ScreenFire showed excellent agreement with research reference typing tests and detection of CIN2+. Risk‐based type results could help guide clinical management of HPV‐positive women. Time‐to‐positive combined with genotyping might be useful. ScreenFire is rapid, mobile, relatively inexpensive and designed for implementation of HPV‐based screening and management, including in lower‐resource settings. Further validation in screening by self‐sampling and practical effectiveness merit evaluation.

Comparison of the ScreenFire and Xpert HPV assays for the detection of human papillomavirus and cervical precancer among women living with HIV in Malawi

“…While the benefits and limitations of the ScreenFire assay have also been discussed in a recent large cross-sectional study, 37 our data add two very important aspects specific to LMIC settings. First, this is the first time, to our knowledge, that the novel redesigned ScreenFire HPV assay has been validated on samples from WLWH, who are known to have generally a higher prevalence of multiple HPV infections.…”

“…The existing 15-type isothermal assay (AmpFire HPV) was recently redesigned for public health use as a 13-type assay (ScreenFire) with four channels based on differential cancer risk: (i) HPV 16, (ii) HPV 18/45, (iii) HPV 31/33/35/52/58, (iv) HPV 39/51/56/59/68. 28,37 This redesigned assay, performed in a single tube, was compared to an AmpliTaq Gold MY09-MY11 PCR-based HPV test on 453 provider-collected samples from Nigeria with very high agreement – the weighted kappa for ScreenFire versus AmpliTaq Gold was 0.90 (95% CI: 0.86-0.93). 28 Recently, ScreenFire was compared to Linear Array and TypeSeq using 2,076 provider-collected samples from the USA, demonstrating excellent clinical performance, with a CIN3+ sensitivity of 94.7% (95% CI: 92.6-96.4) for ScreenFire, 92.3% (95% CI: 89.7-94.3) for Linear Array and 96.0% (95% CI: 93.9-97.6) for TypeSeq.…”

“…28 Recently, ScreenFire was compared to Linear Array and TypeSeq using 2,076 provider-collected samples from the USA, demonstrating excellent clinical performance, with a CIN3+ sensitivity of 94.7% (95% CI: 92.6-96.4) for ScreenFire, 92.3% (95% CI: 89.7-94.3) for Linear Array and 96.0% (95% CI: 93.9-97.6) for TypeSeq. 37 Similarly, ScreenFire was compared to Ampfire, Cobas and SeqHPV genotyping using Chinese samples to satisfy Meijer’s Criteria for clinical endpoint validation, 38 and simulating the VALGENT framework for inter and intra-laboratory validation. 39 In this study using samples from the Chinese Multi-Site Screening Trial (CHIMUST), ScreenFire demonstrated excellent genotype-specific concordance when evaluated for clinical guidance in hierarchical fashion with both the Cobas 4800 and SeqHPV for <CIN2, CIN2, and >CIN3.…”

Comparison of the ScreenFire and Xpert HPV assays for the detection of human papillomavirus and cervical precancer among women living with HIV in Malawi

“…The existing 15-type isothermal assay (AmpFire HPV) was recently redesigned for public health use as a 13-type assay (ScreenFire) with four channels based on differential cancer risk: (i) HPV 16, (ii) HPV 18/45, (iii) HPV 31/33/35/52/58, (iv) HPV 39/51/56/59/68 [28,37]. This redesigned assay, performed in a single tube, was compared to an AmpliTaq Gold MY09-MY11 PCR-based HPV test on 453 provider-collected samples from Nigeria with very high agreement -the weighted kappa for ScreenFire versus AmpliTaq Gold was 0.90 (95% CI 0.86-0.93) [28].…”

“…This redesigned assay, performed in a single tube, was compared to an AmpliTaq Gold MY09-MY11 PCR-based HPV test on 453 provider-collected samples from Nigeria with very high agreement -the weighted kappa for ScreenFire versus AmpliTaq Gold was 0.90 (95% CI 0.86-0.93) [28]. Recently, ScreenFire was compared to Linear Array and TypeSeq using 2,076 provider-collected samples from the USA, demonstrating excellent clinical performance, with a CIN3 + sensitivity of 94.7% (95% CI 92.6-96.4) for ScreenFire, 92.3% (95% CI 89.7-94.3) for Linear Array and 96.0% (95% CI 93.9-97.6) for TypeSeq [37]. Similarly, ScreenFire was compared to Ampfire, Cobas and SeqHPV genotyping using Chinese samples to satisfy Meijer's Criteria for clinical endpoint validation [38], and simulating the VALGENT framework for inter and intra-laboratory validation [39].…”