1. Am J Clin Pathol. 2024 Jun 3;161(6):535-542. doi: 10.1093/ajcp/aqad181.
AmpFire HPV and ScreenFire RS HPV validation trial.
Hou J(1)(2)(3), Belinson JL(4)(5), Du H(1)(2)(3), Li C(1)(2)(3), Zhang
W(1)(2)(3), Zhang L(1)(2)(3), Zhang Y(1)(2)(3), Qu X(1)(2)(3), Wu R(1)(2)(3).
Author information:
(1)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
Shenzhen, China.
(2)Institute of Obstetrics and Gynecology, Shenzhen PKU-HKUST Medical Center,
Shenzhen, China.
(3)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecological Diseases, Shenzhen, China.
(4)Preventive Oncology International, Cleveland, OH, US.
(5)Women’s Health Institute, Lerner College of Medicine, Cleveland Clinic,
Cleveland, OH, US.
OBJECTIVES: The human papillomavirus (HPV) screening assays from Atila
Biosystems, including the new AmpFire (14 type) and ScreenFire RS (13 type),
were subjected to a series of validation tests.
METHODS: We used a set of samples from the Chinese Multi-Site Screening Trial
(previously tested with cobas 4800 and the next-generation SeqHPV) to satisfy
Meijer’s criteria for clinical end-point validation. We selected 556
self-collected specimens composed of 273 high-risk HPV (hrHPV) positives and 283
hrHPV negatives on the cobas 4800 and SeqHPV. Of the 273 hrHPV-positive cases,
108 had a disease end point of cervical intraepithelial neoplasia grade 2 (CIN2)
or higher, including 47 with cervical intraepithelial neoplasia grade 3 (CIN3+)
or higher. We simulated the VALGENT framework for inter- and intralaboratory
validation and evaluated the new 4-channel risk-stratified ScreenFire assay in a
hierarchal fashion.
RESULTS: Both AmpFire and ScreenFire detected 106 (98.1%) of 108 cases with CIN2
or higher, with specificities of 56.7% and 58.1%, respectively. Intralaboratory
concordance for 2 runs of AmpFire and ScreenFire was 95.13% and 96.03%,
respectively, for overall hrHPV types and 99.10% and 99.46%, respectively, for
HPV 16. The interlaboratory concordance of AmpFire and ScreenFire was 93.68% and
94.04% for overall hrHPV and 98.92% and 99.28%, respectively, for HPV 16. Other
genotype correlation percentages were similarly high, with κs ranging from 0.86
to 0.94. The ScreenFire RS assay demonstrated excellent “genotype-specific
concordance” when evaluated for “clinical guidance” in a hierarchal fashion
(noting only the highest risk channel) with both the cobas 4800 and SeqHPV for
less than CIN2, CIN2, and CIN3 or higher.
CONCLUSIONS: The excellent intra- and interlaboratory reproducibility and the
established clinical performance, together with the platforms’ simplicity, make
these assays particularly applicable to low-resource settings.
© The Author(s) 2024. Published by Oxford University Press on behalf of American
Society for Clinical Pathology. All rights reserved. For commercial re-use,
please contact reprints@oup.com for reprints and translation rights for
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DOI: 10.1093/ajcp/aqad181
PMID: 38365314 [Indexed for MEDLINE]
2. Infect Agent Cancer. 2024 Nov 29;19(1):59. doi: 10.1186/s13027-024-00622-2.
Analytic performance of ScreenFire HPV RS assay Zebra BioDome format and its potential for large-scale population HPV screening.
Wang J(1)(2), Imade G(3), Akanmu AS(4), Musa J(5)(6)(3), Anorlu R(7), Zheng
Y(5)(6), Garcia-Bedoya O(8), Sanchez GI(9), Belinson J(10), Kim K(5), Maiga
M(5)(6), Gursel DB(11), Sagay AS(3), Ogunsola FT(12), Murphy RL(13)(14), Hou
L(15)(16).
Author information:
(1)Department of Preventive Medicine, Division of Cancer Epidemiology and
Prevention, Feinberg School of Medicine, Northwestern University, 680 N Lake
Shore Dr, Suite 1400, Chicago, IL, 60611, USA. j-wang4@northwestern.edu.
(2)Center for Global Oncology, Robert J. Havey MD Institute for Global Health,
Feinberg School of Medicine, Northwestern University, Chicago, USA.
j-wang4@northwestern.edu.
(3)Department of Obstetrics and Gynecology, College of Health Sciences,
University of Jos, Jos, Nigeria.
(4)Department of Hematology and Blood Transfusion, College of Medicine, Lagos
University Teaching Hospital, University of Lagos, Lagos, Nigeria.
(5)Department of Preventive Medicine, Division of Cancer Epidemiology and
Prevention, Feinberg School of Medicine, Northwestern University, 680 N Lake
Shore Dr, Suite 1400, Chicago, IL, 60611, USA.
(6)Center for Global Oncology, Robert J. Havey MD Institute for Global Health,
Feinberg School of Medicine, Northwestern University, Chicago, USA.
(7)Department of Obstetrics and Gynecology, College of Medicine, University of
Lagos, Lagos, Nigeria.
(8)Division of Academic Internal Medicine and Geriatrics, Institute for Minority
Health Research, University of Illinois College of Medicine, Chicago, USA.
(9)Infection and Cancer Group, Faculty of Medicine, University of Antioquia,
Medellin, Colombia.
(10)Women’s Health Institute, Cleveland Clinic, Cleveland, USA.
(11)Department of Pathology, Feinberg School of Medicine, Northwestern
University, Chicago, IL, USA.
(12)Department of Medical Microbiology, College of Medicine, University of
Lagos, Lagos, Nigeria.
(13)Division of Infectious Diseases, Department of Medicine, Feinberg School of
Medicine, Northwestern University, Chicago, USA.
(14)Robert J. Havey MD Institute for Global Health, Northwestern University,
Chicago, USA.
(15)Department of Preventive Medicine, Division of Cancer Epidemiology and
Prevention, Feinberg School of Medicine, Northwestern University, 680 N Lake
Shore Dr, Suite 1400, Chicago, IL, 60611, USA. l-hou@northwestern.edu.
(16)Center for Global Oncology, Robert J. Havey MD Institute for Global Health,
Feinberg School of Medicine, Northwestern University, Chicago, USA.
l-hou@northwestern.edu.
BACKGROUND: Easy-to-use, rapid, scalable, high-throughput, and cost-effective
HPV tests are urgently needed for low-resource settings. Atila Biosystems’
high-throughput, cost-effective, and clinically validated ScreenFire HPV Risk
Stratification (RS) assay identifies 13 high risk HPV (hrHPV) in 4 groups based
on their oncogenic risk (i.e., HPV16, HPV18/45, HPV31/33/35/52/58, and
HPV51/59/39/56/68). The current standard format is subject to laboratory
contamination, which is common for any molecular PCR test. To overcome this
drawback, Atila has recently upgraded it into an innovative, contamination-free
Zebra BioDome format. The contamination-free feature makes this novel assay
format more suitable for large-scale community- and population-based cervical
screening. This study evaluated the analytical performance of the Zebra BioDome
format.
METHODS: We conducted a study to test the analytical performance of Zebra
Biodome format in comparison to the results of using the ScreenFire HPV RS assay
standard format on Biorad CFX-96 real-time PCR instrument. We used overall
agreement rate and unweighted kappa value to compare the performance.
RESULTS: The overall agreement for detection of hrHPV was 96.0% with unweighted
kappa value 0.94 (95% confidence interval: 0.90-0.98). The agreement rates
between hrHPV genotype 16 and risk stratification genotype group (HPV18/45,
HPV31/33/35/52/58, and HPV51/59/39/56/68) were all > 97.5%.
CONCLUSION: The innovative ScreenFire HPV RS assay Zebra BioDome format produced
highly concordant results with the standard format. The shared features by the
two assay formats, such as easy-to-use, high throughput, cost-appropriate, and
no requirements for DNA extraction. The unique contamination-prevention feature
along with no requirement of preparation of reagents make the Zebra BioDome
format more suitable for large-scale HPV screening to reduce global cervical
cancer burden.
© 2024. The Author(s).
DOI: 10.1186/s13027-024-00622-2
PMCID: PMC11606105
PMID: 39614290
Conflict of interest statement: Declarations. Ethics approval and consent to
participate: This study is covered under IRB approval (STU00207051) at
Northwestern University. Consent for publication: Not applicable. Competing
interests: The authors declare no competing interests.
3. Infect Agent Cancer. 2025 Apr 30;20(1):28. doi: 10.1186/s13027-025-00651-5.
Analytical performance of the ScreenFire HPV RS Zebra BioDome assay on four different qPCR platforms.
Wang J(1)(2), Imade G(3), Akanmu AS(4), Musa J(3), Anorlu R(5), Zheng Y(6)(7),
Joyce B(6)(7), Adewole I(8), Morhason-Bello IO(9), Belinson J(10), Maiga
M(6)(7), Gursel DB(11), Sagay AS(3), Ogunsola FT(12), Murphy RL(13)(14), Hou
L(15)(16).
Author information:
(1)Department of Preventive Medicine, Division of Cancer Epidemiology and
Prevention, Feinberg School of Medicine, Northwestern University, Chicago, USA.
j-wang4@northwestern.edu.
(2)Center for Global Oncology, Robert J. Havey Institute for Global Health,
Feinberg School of Medicine, Northwestern University, Chicago, USA.
j-wang4@northwestern.edu.
(3)Department of Obstetrics and Gynecology, College of Health Sciences,
University of Jos, Jos, Nigeria.
(4)Department of Hematology and Blood Transfusion, College of Medicine, Lagos
University Teaching Hospital, University of Lagos, Lagos, Nigeria.
(5)Department of Obstetrics and Gynecology, College of Medicine, University of
Lagos, Lagos, Nigeria.
(6)Department of Preventive Medicine, Division of Cancer Epidemiology and
Prevention, Feinberg School of Medicine, Northwestern University, Chicago, USA.
(7)Center for Global Oncology, Robert J. Havey Institute for Global Health,
Feinberg School of Medicine, Northwestern University, Chicago, USA.
(8)HPV Research Consortium, College of Medicine, University of Ibadan, Ibadan,
Nigeria.
(9)Department of Obstetrics and Gynaecology, and HPV Research Consortium,
College of Medicine, University of Ibadan, Ibadan, Nigeria.
(10)Women’s Health Institute, Cleveland Clinic, and Preventive Oncology
International, Inc., Cleveland, USA.
(11)Department of Pathology, Feinberg School of Medicine, Northwestern
University, Chicago, USA.
(12)Department of Medical Microbiology, College of Medicine, University of
Lagos, Lagos, Nigeria.
(13)Division of Infectious Diseases, Department of Medicine, Feinberg School of
Medicine, Northwestern University, Chicago, USA.
(14)Robert J. Havey MD Institute for Global Health, Northwestern University,
Chicago, USA.
(15)Department of Preventive Medicine, Division of Cancer Epidemiology and
Prevention, Feinberg School of Medicine, Northwestern University, Chicago, USA.
l-hou@northwestern.edu.
(16)Center for Global Oncology, Robert J. Havey Institute for Global Health,
Feinberg School of Medicine, Northwestern University, Chicago, USA.
l-hou@northwestern.edu.
OBJECTIVES: Cervical cancer is one of the most frequently diagnosed cancers and
a leading cause of cancer-related deaths in women in low- and middle-income
countries (LMICs), accounting for nearly 85% of the global cervical cancer
burden. High-risk human papillomavirus (hrHPV) infection is the main cause of
cervical cancer. Easy-to-use, rapid, scalable, high-throughput, and
cost-effective HPV tests are urgently needed for low-resource settings. Atila
Biosystems’ clinically validated ScreenFire HPV Risk Stratification (RS) assay
identifies 13 hrHPV in 4 groups based on their oncogenic risk (i.e., HPV16,
HPV18/45, HPV31/33/35/52/58, and HPV51/59/39/56/68). While the current standard
format is subject to laboratory contamination Atila has developed an innovative,
contamination-preventive Zebra BioDome format. Recently we published the
analytical performance of ScreenFire RS Zebra BioDome on the BioRad CFX-96
real-time PCR instrument. This current study evaluated its analytical
performance on three additional qPCR platforms: Atila Portable iAMP-PS96, Atila
Powergene9600 Plus, and Thermo Fisher Quantstudio-7.
METHODS: We tested 173 DNA samples from Nigerian women with cervical cancer.
These samples were tested simultaneously using the ScreenFire HPV Zebra BioDome
assay (M5FHPV-96) on four different real-time PCR machines (Atila portable
iAMP-PS96, Atila Powergene9600 Plus, Thermo Fisher QuantStudio-7, and BioRad
CFX-96). We used overall agreement rate and unweighted kappa values to compare
different platforms.
RESULTS: The overall agreement for detection of hrHPV using Atila portable
iAMP-PS96 was 96.5% with kappa value 0.95 (95% confidence interval: 0.91-0.99)
compared to Thermo Fisher QuantStudio-7, and 97.1% with kappa value 0.96 (95%
confidence interval: 0.92-0.99) compared to BioRad CFX-96. For genotype HPV16
and risk stratification (RS) genotype groups (HPV18/45, HPV31/33/35/52/58, and
HPV51/59/39/56/68) agreement rates were all > 98.3%. For Atila Powergene9600
Plus the overall agreement was 98.8% with a kappa value of 0.98 (95% confidence
interval: 0.96-1.0) compared to Thermo Fisher QuantStudio-7, and 96.5% with a
kappa value of 0.96 (95% confidence interval: 0.94-0.99) compared to BioRad
CFX-96. The agreements for the HPV16 and RS genotype groups (HPV18/45,
HPV31/33/35/52/58, and HPV39/51/56/59/68) were at least 98.3%.
CONCLUSION: The novel ScreenFire HPV Zebra BioDome format produced highly
concordant hrHPV positivity and RS genotype results on all four qPCR platforms.
The data suggests that this innovative technology has the potential to improve
HPV testing uptake in low-resource settings without further investment in
purchasing new equipment.
© 2025. The Author(s).
DOI: 10.1186/s13027-025-00651-5
PMCID: PMC12042583
PMID: 40307888
Conflict of interest statement: Declarations. Ethics approval and consent to
participate: This study is covered under IRB approval at Northwestern University
(STU00207051), Jos University (JUTH/DCS/ADM/127/XXVII/630) and Lagos University
(CMUL/HREC/01/18/327). Consent for publication: Not applicable. Competing
interests: The authors declare no competing interests.