With great progress in HPV assay development over the past 5-7 years, and especially PCR based assays, BGI Shenzhen (the world’s largest genomic sequencing facility) asked the question if sequencing technology can move the science and the POI screening paradigm even further. With the new bench-top next generation sequencing instruments currently available such as the Ion Torrent Personal Genome Machine (PGM, Life Technologies, Carlsbad CA, USA ) and MiSeq (Illumina, San Diego, CA, USA), fast progress in DNA sequencing technology has allowed a substantial reduction in costs along with improved accuracy and throughput. BGI in collaboration with POI believed it would be possible that by using next generation sequencing (NGS) technology, we could drive the costs lower and the applicability even wider due to the cost and transportability of the latest in NGS technology.To achieve this goal, we designed a high throughput HPV genotyping assay based on multiplex PCR and next generation sequencing. We used PGM and MiSeq, and optimized the HPV positive cutoff of this new assay and then validated the assay on population based self-collected samples from SHENCCAST II. The result was a high throughput, highly sensitive, low cost per assay. The sensitivity for ≥CIN 2 and ≥CIN 3 of the self-sampling specimens tested by the new NGS HR-HPV genotyping assay (SEQHPV) was similar to that of direct sampling specimens tested by HC2 (p>0.05)，but the specificity for ≥CIN 2 and ≥CIN 3 of SEQHPV was significantly higher than HC2 (p<0.01). This work “Development and validation of a new HPV genotyping assay based on next generation sequencing”, was Accepted November 2013, for publication in the Am J. Clinical Pathology. We hope to continue this work to further improve assay specificity by incorporating the detection of viral methylation.
REFERENCES
1. PLoS One. 2020 May 1;15(5):e0232117. doi: 10.1371/journal.pone.0232117.
eCollection 2020.
The application of BMRT-HPV viral load to secondary screening strategies for cervical cancer.
Duan L(1)(2), Du H(1)(2), Wang C(1)(2), Huang X(1)(2), Qu X(3), Shi B(4), Liu
Y(5), Zhang W(6), Duan X(7), Wei L(8), Belinson JL(9)(10), Wu R(1)(2).
Author information:
(1)Department of Gynecology and Obstetrics, Peking University Shenzhen Hospital,
Shenzhen, China.
(2)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecological Diseases, Shenzhen, PR,China.
(3)Sanming Project of Medicine in Shenzhen Peking University Shenzhen Hospital,
Shenzhen, China.
(4)Department of Gynecology and Obstetrics, The Second Hospital of Hebei Medical
University, Hebei, China.
(5)Department of Gynecology and Obstetrics, Huashan Hospital North, Fudan
University, Shanghai, China.
(6)Department of Gynecology and Obstetrics, Zhongnan Hospital of Wuhan
University, Wuhan, China.
(7)Department of Gynecology and Obstetrics, Capital Medical University Beijing
Tongren Hospital, Beijing, PR, China.
(8)Department of Gynecology and Obstetrics, Peking University People’s Hospital,
Beijing, PR, China.
(9)Preventive Oncology International, Cleveland Heights, OH, United States of
America.
(10)Women’s Health Institute, Cleveland Clinic, Cleveland, OH, United States of
America.
OBJECTIVE: Evaluate the significance of BMRT HPV assay viral load and its
performance for secondary screening.
METHODS: BMRT-HPV reports type-specific viral loads/10,000 cells. We tested
1,495 physician collected, stored specimens from Chinese Multiple-center
Screening Trial (CHIMUST), that were positive by Cobas, SeqHPV, and/or Cytology
(≥LSIL); and 2,990 age matched, negatives in a nested case control study. We
explored the relationship between BMRT HR-HPV viral load and cervical lesions,
determined alternative CIN2+ cut-points by ROC curve, and evaluated BMRT HR-HPV
for primary / secondary cervical cancer screening.
RESULTS: The viral loads of HPV16/18, 12 other subtypes HR-HPV and 14 HR-HPV
were statistically different in all grades of cervical lesions (P<0.05, among
which HPV16, 33 and 58 showed the strongest relationship (P<0.01). The viral
load of HR-HPV also increased with the grade of cervical lesions (P<0.05). The
sensitivity for CIN2+ and CIN3+ of BMRT was comparable to Cobas (92.6% vs 94.3%,
100% vs 100%, P>0.05), specificity was higher than Cobas (84.8% vs 83.3%, 83.5%
vs 82.0%, P<0.001). When using HPV16/18 viral load(log cut-point ≥3.2929), plus
the viral-load of 12 other subtypes (log cut-point ≥3.9625) as secondary triage,
compared with Cobas HPV16/18+ plus cytology ≥ASC-US as triage, the sensitivities
for CIN2+ and CIN3+ were similar (P>0.05). However, the BMRT HR-HPV viral load
combined with subtypes did not require cytology.
CONCLUSION: BMRT is as sensitive as Cobas4800 for primary cervical cancer
screening. BMRT HR-HPV viral load combined with subtypes can be used as a
secondary strategy for cervical cancer screening, especially for areas with
insufficient cytological resources.
DOI: 10.1371/journal.pone.0232117
PMCID: PMC7194433
PMID: 32357165 [Indexed for MEDLINE]
Conflict of interest statement: JLB is affiliated with Preventive Oncology
International, Inc, OH. There are no patents, products in development or
marketed products to declare. This does not alter our adherence to PLOS ONE
policies on sharing data and materials. The authors have declared that no other
competing interests exist
2. Infect Agent Cancer. 2020 Oct 23;15:65. doi: 10.1186/s13027-020-00328-1.
eCollection 2020.
Evaluation of an isothermal amplification HPV detection assay for primary cervical cancer screening.
Zhang W(#)(1)(2), Du H(#)(1)(2), Huang X(1)(2), Wang C(1)(2), Duan X(3), Liu
Y(4), Shi B(5), Zhang W(6), Qu X(7), Wei L(8), Schiffman M(9), Belinson
JL(10)(11), Wu R(1)(2).
Author information:
(1)Peking University Shenzhen Hospital, Shenzhen, 518036 P. R. China.
(2)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecological diseases, Shenzhen, P. R. China.
(3)Capital Medical University Beijing Tongren Hospital, Beijing, P. R. China.
(4)Fudan University Huanshan Hospital, Shanghai, P. R. China.
(5)The second Hospital of Hebei Medical University, Shijiazhuang, P. R. China.
(6)Wuhan University Zhongnan Hospital, Wuhan, P. R. China.
(7)Expert in Three Engineering Office, Shenzhen Maternal of Peking University
Shenzhen Hospital, Shenzhen, 518036 China.
(8)Department of Obstetrics and Gynecology, Peking University People’s Hospital,
Beijing, P. R. China.
(9)National Cancer Institute, Division of Epidemiology and Genetics, Bethesda,
USA.
(10)Women’s Health Institute, Cleveland Clinic, Cleveland, OH USA.
(11)PPreventive Oncology International, Inc., Shaker Heights, USA.
(#)Contributed equally
OBJECTIVE: The aim of this research was to evaluate independently the
performance of a new isothermal amplification assay for cervical cancer
screening compared to two previously validated PCR-based assays and histologic
endpoints.
METHODS: This is a sub-study from the Chinese multi-center screening trial
(CHIMUST). The self-collected and clinician-collected specimens stored in
PreservCyt at - 4 °C from 6042 women with complete data were tested with the
AmpFire assay. These specimens had been previously tested with Cobas and SeqHPV
assays. In the primary study all patients with an abnormal test were referred to
colposcopy where all had directed and/or random biopsies plus ECC. No additional
patients were called back based on the AmpFire results.
RESULTS: 6042/6619 women had complete data (mean age 44.1). There were 57 cases
of CIN 2, 35 cases of CIN 3 and 2 cancers. The sensitivity for CIN2+ and CIN3+
were similar among the three assays (both direct and self-collected). For the
specificities in all categories (CIN2+/CIN3+ and self and direct collection),
isothermal amplification assay was either equal to or more specific than Cobas
but consistently less specific than SeqHPV.
CONCLUSION: The AmpFire HPV assay showed similar sensitivity to Cobas and SeqHPV
for CIN2+ and CIN3+ on both self and clinician-collections (P>0.05), with good
specificity. The speed, low cost, and simplicity of this assay will make it
particularly suited for low and middle resource settings. Its accuracy with
self-collection makes it applicable for mass screening programs.
© The Author(s) 2020.
DOI: 10.1186/s13027-020-00328-1
PMCID: PMC7583687
PMID: 33110442
Conflict of interest statement: The authors declare no conflict of interest.
3. Zhonghua Fu Chan Ke Za Zhi. 2020 Oct 25;55(10):708-715. doi:
10.3760/cma.j.cn112141-20200325-00266.
[Evaluation of the effectiveness of BMRT-HPV for cervical cancer screening].
[Article in Chinese; Abstract available in Chinese from the publisher]
Duan LF(1), Du H(1), Wang C(1), Huang X(1), Qu XF(1), Duan XZ(2), Liu Y(3), Shi
B(4), Zhang W(5), Wei LH(6), Belinson L(7), Wu RF(1).
Author information:
(1)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
Shenzhen Key Laboratory on Technology for Early Diagnosis of Major Gynecological
Diseases, Shenzhen 518036, China.
(2)Department of Obstetrics and Gynecology, Capital Medical University Beijing
Tongren Hospital, Beijing 100730, China.
(3)Department of Obstetrics and Gynecology, Huashan Hospital, Fudan University,
Shanghai 200003, China.
(4)Department of Obstetrics and Gynecology, the Second Hospital of Hebei Medical
University, Shijiazhuang 050005, China.
(5)Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan
University, Wuhan 430071, China.
(6)Department of Obstetrics and Gynecology, Peking University People’s Hospital,
Beijing 100044, China.
(7)Preventive Oncology International, Cleveland Heights, OH, United States of
America, 44101.
Objective: Evaluation of the clinical value of the BioPerfectus multiplex real
time (BMRT)-HPV for cervical cancer screening. Methods: Physician-collected
specimens of 1 495 women who were positive of Cobas 4800 HPV (Cobas-HPV), HPV
genotyping based on SEQ uencing (SEQ-HPV), and (or) cytology ≥low grade squamous
intraepithelial lesion (LSIL) in the primary screening of Chinese
Multiple-center Screening Trial (CHIMUST), and 2 990 women selected from those
who were negative of primary screening in the same project through nested
control randomization with age-matching were tested for BMRT-HPV, which reported
type-specific viral loads/10 000 cells in each specimen. With comparing to
Cobas-HPV results and taking cervical histopathological diagnosis as the
endpoint, the concordance of high-risk (HR)-HPV subtypes among the three assays
was explored ,and the sensitivity and specificity of BMRT-HPV for cervical
cancer screening were evaluated. Results: (1) The overall agreenment of HR-HPV
subtypes between BMRT-HPV and Cobas-HPV, or SEQ-HPV test sample was 94.8%,
94.4%, with Kappa values 0.827, 0.814. (2) The sensitivity and specificity for
cervical intraepithelial neoplasia (CIN) Ⅱ+ of BMRT-HPV, Cobas-HPV and SEQ-HPV
were 92.62%, 94.26%, 93.44% and 84.67%, 83.25%, 82.76%, respectively. There were
no significant difference in sensitivity among the three HPV assays (all
P>0.05), but the specificity of BMRT-HPV for CIN Ⅱ+ was higher than those of
Cobas-HPV and SEQ-HPV (P<0.01). The sensitivity for CIN Ⅲ+ of three HPV assays
were all 100.00%, and the specificity for CIN Ⅲ+ of BMRT-HPV was higher than
those of Cobas-HPV and SEQ-HPV (83.40% vs 81.95%, 83.40% vs 81.50%; P<0.01). The
number of pathological examinations of colposcopy for cervical biopsy detected
in 1 case of CIN Ⅱ+ or CIN Ⅲ+ in BMRT-HPV was less than those in Cobas-HPV and
SEQ-HPV (P<0.01). When using HPV 16/18 + cytology ≥atypical squamous cell of
undetermined signification (ASCUS) to triage HPV positive women among three
assays, there was no different in the sensitivities of detecting CIN Ⅱ+ and CIN
Ⅲ+ (P>0.05). The specificity BMRT-HPV was slightly higher than those in
Cobas-HPV or SEQ-HPV (all P<0.05), and the colposcopy referral rate was lower
than those in Cobas-HPV and SEQ-HPV (all P<0.05). Conclusions: BMRT-HPV is as
sensitive as Cobas-HPV or SEQ-HPV for primary cervical cancer screening, and has
higher specificity. Therefore it could be used as a primary screening method for
cervical cancer, which is worthy of clinical application.
Publisher: 目的： 评价核酸分型定量法HPV检测（BMRT-HPV）用于子宫颈癌筛查的临床价值。 方法：
采用巢式抽样法，在2016年9月—2018年1月中国子宫颈癌筛查多中心研究（CHIMUST）项目5个筛查点的8
856例妇女中，选取其中医生取样或自取样标本中任一检测方法HPV阳性或HPV阴性但细胞学结果≥低级别鳞状上皮内病变（LSIL）者共1
495例行BMRT-HPV检测，同时按照1∶2比例抽取年龄和参加筛查时间相匹配的HPV与细胞学结果均为阴性的2
990例为对照。CHIMUST项目中对任一HPV阳性妇女回叫行阴道镜下子宫颈病灶四象限定点活检+随机活检+子宫颈管搔刮术方案子宫颈活检，以医生取样标本的基于实时PCR技术的Cobas
4800
HPV检测（Cobas-HPV）、基于第2代基因测序技术的HPV分型检测（SEQ-HPV）为对照，以子宫颈活检病理诊断为“金标准”，分析BMRT-HPV检测的高危型HPV（HR-HPV）亚型与Cobas-HPV、SEQ-HPV检测的一致性，并比较3种HPV检测方法对子宫颈上皮内瘤变（CIN）Ⅱ及以上级别病变（CIN
Ⅱ+）、CIN Ⅲ及以上级别病变（CIN Ⅲ+）的筛查效率。 结果：
（1）BMRT-HPV检测方法分别与Cobas-HPV、SEQ-HPV检测方法比较，检测HR-HPV亚型整体的一致性分别为94.8%、94.4%，Kappa值分别为0.827、0.814。（2）BMRT-HPV、Cobas-HPV、SEQ-HPV
3种方法对CINⅡ+筛查的敏感度分别为92.62%、94.26%、93.44%，两两比较，差异均无统计学意义（P>0.05）；特异度分别为84.67%、83.25%、82.76%，BMRT-HPV检测对CINⅡ+筛查的特异度显著高于Cobas-HPV、SEQ-HPV检测（P<0.01）。3种HPV检测方法对CIN
Ⅲ+筛查的敏感度均达到100.00%，BMRT-HPV检测对CIN
Ⅲ+筛查的特异度也显著高于Cobas-HPV、SEQ-HPV检测（分别为83.40%、81.95%、81.50%，P<0.01）。检出1例CINⅡ+或CIN
Ⅲ+所需要行阴道镜下子宫颈活检病理检查的患者数，BMRT-HPV检测显著少于Cobas-HPV和SEQ-HPV检测（P<0.01）。Cobas-HPV、SEQ-HPV、BMRT-HPV检测初筛阳性者分别以HPV
16和（或）18型（HPV
16/18型）阳性联合细胞学结果≥未明确诊断意义的不典型鳞状上皮细胞（ASCUS；即为方案一、二、三）进行二次分流，3种筛查方案对CIN Ⅱ+、CIN
Ⅲ+筛查的敏感度分别比较均无显著差异（P>0.05），但方案三（即BMRT-HPV初筛阳性者）筛查CIN Ⅱ+、CIN
Ⅲ+的特异度显著高于方案一（即Cobas-HPV初筛阳性者）和方案二（即SEQ-HPV初筛阳性者；P<0.05），而且阴道镜转诊率方案三也显著低于方案一、二（P<0.05）。
结论： BMRT-HPV检测筛查子宫颈癌具有与Cobas-HPV、SEQ-HPV检测相似的敏感度，但特异度更高，可作为子宫颈癌的初筛方法，值得临床推广使用。.
DOI: 10.3760/cma.j.cn112141-20200325-00266
PMID: 33120484 [Indexed for MEDLINE]
4. Infect Agent Cancer. 2020 Nov 30;15(1):72. doi: 10.1186/s13027-020-00333-4.
An evaluation of solid versus liquid transport media for high-risk HPV detection and cervical cancer screening on self-collected specimens.
Du H(1), Duan X(2), Liu Y(3), Shi B(4), Zhang W(5), Wang C(1), Qu X(1), Bao
J(1), Li J(6), Zhao C(6), Jiang J(4), Liu J(3), Wu K(5), Xiao A(1), Duan L(1),
Huang X(1), Bian S(1), Zhang L(1), Luo H(1), Wei L(7), Belinson JL(8), Wu R(9);
CHIMUST group.
Author information:
(1)Peking University Shenzhen Hospital, Shenzhen, PR China.
(2)Capital Medical University Beijing Tongren Hospital, Beijing, PR China.
(3)Fudan University Huashan Hospital North, Shanghai, PR China.
(4)The second Hospital of Hebei Medical University, Shijiazhuang, PR China.
(5)Wuhan University Zhongnan Hospital, Wuhan, PR China.
(6)Peking University People’s Hospital, Beijing, PR China.
(7)Peking University People’s Hospital, Beijing, PR China. weilhpku@163.com.
(8)Preventive Oncology International Inc, and the Cleveland Clinic, Cleveland,
OH, USA. jib@poiinc.org.
(9)Peking University Shenzhen Hospital, Shenzhen, PR China. wurfpush@126.com.
BACKGROUND: The solid transport media is a small size card that allows fast,
easy DNA extraction from a variety of biological samples. In 2016 we developed a
solid media transport card; for that pilot study to control the self-collection
we used a pseudo-self-collection technique. The current study expands this prior
work using true self-collections and only the POI card, and aims to evaluate the
solid media transport card to detect HR-HPV in self-samples compared to liquid
transport media.
METHODS: Ten thousand eight hundred eighty-five women between the ages of 30-59
with no screening for 3 years were enrolled. The self-collected sample was first
applied to a new solid media transport card (Labeled as SC) then the brush
placed in 6 ml ThinPrep liquid (Labeled as SL). Then a physician collected a
direct endocervical specimen into ThinPrep liquid (Labeled as DL). Samples were
tested with Cobas 4800 and the SeqHPV NGS assay for HR-HPV. Patients positive on
any test were recalled for colposcopy and biopsy.
RESULTS: Ten thousand three hundred thirty-nine participants had complete data.
The mean age was 43.9 years. CIN 2+ rates were 1.4% (142/10339). The agreement
in HPV detection between the two different self-sample collection media was also
good (Cobas HPV kappa = 0.86; SeqHPV kappa = 0.98). Tested with Cobas, the
sensitivity of Cobas-SL and Cobas-SC for CIN 2+ was95.07 and 94.37%; and for
CIN3+ was 96.30, 96.30% respectively. The specificity of Cobas-SC, and Cobas-SL
for CIN2+ was 88.74 and 87.35%; for CIN3 was 88.04and 86.65% respectively.
Tested with SeqHPV, the sensitivity for CIN2+ of Seq-SC and Seq-SL was 95.77 and
96.48%; for CIN3+, both the SC and SL specimens had a sensitivity of 100%. The
specificity for CIN2+ of Seq-SC and Seq-SL was 89.54 and 89.53%; for CIN3+ was
88.84,88.82% respectively. For both HR-HPV assays, the sensitivities were
similar for the two self-sample media (SC vs SL, p = 1.00).
CONCLUSIONS: The solid transport card for collecting vaginal self-samples as
accurate as liquid transport media assayed by two different PCR based HR-HPV
tests. The solid transport media is a suitable medium for collecting and storing
vaginal self-samples.
DOI: 10.1186/s13027-020-00333-4
PMCID: PMC7706049
PMID: 33292341
Conflict of interest statement: The funders had no role in study design, in the
collection, analysis and interpretation of data, and decision to publish, or
preparation of the manuscript. All the authors declare no conflicts of
interests.
5. J Low Genit Tract Dis. 2021 Jan 1;25(1):22-26. doi:
10.1097/LGT.0000000000000577.
Evaluation of Cobas HPV and SeqHPV Assays in the Chinese Multicenter Screening Trial.
Du H, Duan X(1), Liu Y(2), Shi B(3), Zhang W(4), Wang C, Qu X(5), Li J(6), Zhao
C(6), Liu J(2), Jiang J(3), Jin H(7), Li H(8), Xiao A, Bao J, Duan L, Huang X,
Luo H(5), Bian S(5), Zhang L(5), Wei L(6), Belinson JL(9), Wu R.
Author information:
(1)Capital Medical University Beijing Tongren Hospital, Beijing, China.
(2)Fudan University Huanshan Hospital North, Shanghai, China.
(3)The second Hospital of Hebei Medical University, Shijiazhuang, China.
(4)Wuhan University Zhongnan Hospital, Wuhan, China.
(5)Peking University Shenzhen Hospital, Shenzhen, China.
(6)Peking University People’s Hospital, Beijing, China.
(7)Wushenqi People’s Hospital, Inner Mongolia, China.
(8)Pingxiang Ganxi Cancer Hospital, Jiangxi Province, China.
(9)Preventive Oncology International, Inc, and the Cleveland Clinic, Cleveland,
OH.
OBJECTIVE: The aim of the study was to evaluate the Cobas 4800 Assay and the
SeqHPV Assay with self (S) and direct (D) cervical samples in the Chinese
Multicenter Screening Trial (CHIMUST).
MATERIALS AND METHODS: The CHIMUST is a large population-based multicenter
clinical trial, and 10,885 women aged 30-59 years from 15 sites in 7 provinces
with no cervical cancer screening for 3 years were eligible. All participating
women contributed one self-collected sample (S) and 1 physician-collected
endocervical sample (DL). The self-collected sample was first applied to the
solid media transport card (SS), and then, the brush placed in 6 mL of
ThinPrepSolution (SL). All samples were tested with Cobas 4800 and SeqHPV
high-risk HPV assays. Patients human papillomavirus positive (self or direct)
were recalled for colposcopy and biopsies.
RESULTS: A total of 10,399 women had complete data. The mean age was 43.9 years.
A total of 1.4% (142/10,399) had cervical intraepithelial neoplasia (CIN) 2+ and
0.5% (54/10,339) had CIN 3+. In the liquid specimens, the overall HPV infection
rates were 10.8% for Cobas and 10.9% for SeqHPV in D sample, and 13.7% for Cobas
and 11.6% for SeqHPV in SL sample, respectively. The sensitivity of Cobas-DL,
Cobas-SL, SeqHPV-DL, and SeqHPV-SL for CIN 2+ was 95.07%, 95.07%, 94.33%, and
96.48%, respectively. The specificity of Cobas-DL, Cobas-SL, SeqHPV-DL, and
SeqHPV-SL for CIN 2+ was 90.38%, 87.35%, 90.21%, and 89.53%, respectively. There
were no differences in sensitivity when applying the 2 assays to both self- and
directly collected samples in liquid transport media (p > .05).
CONCLUSIONS: Both Cobas and SeqHPV screening assays using both self-collected
and directly endocervical collected specimens demonstrate similar sensitivity
for the detection of CIN 2+ and CIN 3+.
Copyright © 2020, ASCCP.
DOI: 10.1097/LGT.0000000000000577
PMID: 33347045 [Indexed for MEDLINE]
Conflict of interest statement: The authors have declared they have no conflicts
of interest.
6. Gynecol Oncol. 2021 Aug;162(2):322-330. doi: 10.1016/j.ygyno.2021.05.014. Epub
2021 May 29.
Evaluation of p16(INK4a) immunocytology and human papillomavirus (HPV) genotyping triage after primary HPV cervical cancer screening on self-samples in
China.
Song F(1), Belinson JL(2), Yan P(1), Huang X(1), Wang C(1), Du H(3), Qu X(4), Wu
R(5).
Author information:
(1)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
Shenzhen 518036, PR China; Institute of Obstetrics and Gynecology, Shenzhen
PKU-HKUST Medical Center, Shenzhen 518036, PR China; Shenzhen Key Laboratory on
Technology for Early Diagnosis of Major Gynecological Diseases, Shenzhen 518036,
PR China.
(2)Preventive Oncology International, Cleveland Heights, OH, USA; The Women’s
Health Institute, Cleveland Clinic, Cleveland, OH, USA.
(3)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
Shenzhen 518036, PR China; Institute of Obstetrics and Gynecology, Shenzhen
PKU-HKUST Medical Center, Shenzhen 518036, PR China; Shenzhen Key Laboratory on
Technology for Early Diagnosis of Major Gynecological Diseases, Shenzhen 518036,
PR China. Electronic address: duhui_107108@163.com.
(4)Sanming Project of Medicine in Shenzhen, Peking University Shenzhen Hospital,
Shenzhen 518036, PR China. Electronic address: steve1005@icloud.com.
(5)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
Shenzhen 518036, PR China; Institute of Obstetrics and Gynecology, Shenzhen
PKU-HKUST Medical Center, Shenzhen 518036, PR China; Shenzhen Key Laboratory on
Technology for Early Diagnosis of Major Gynecological Diseases, Shenzhen 518036,
PR China. Electronic address: wurfpush@126.com.
OBJECTIVE: Self-sampling for human papillomavirus (HPV) testing is an effective
option to increase the cervical screening coverage. How to best triage
HPV-positive self-samples remains controversial. Here, we evaluated the
performance of a novel p16INK4a immunocytology approach (p16) and HPV genotyping
in triaging HPV-positive self-samples.
METHODS: A cohort of 73699 women were screened via SeqHPV assay on self-samples.
HPV-positive women who met any sequential positive result of HPV16/18 or VIA or
p16 were referred for colposcopy. A triage strategy was considered favorable if
the NPV for CIN3+ was ≥98%, combined with an improvement of sensitivity and
specificity in comparison to the comparator, being the ‘ASC-US+’ triage and the
guideline strategy (HPV16/18+ or ASC-US+).
RESULTS: A total of 3510 HPV-positive women were included, 422 (12.0%) CIN2+ and
247 (7.0%) CIN3+ were identified. The positivity of p16 and ASC-US+ were 36.3%
and 22.2%, respectively. p16 was more sensitive and less specific than ASC-US+
(P < 0.0001). However, when combined p16 with cytology or genotypes, two triage
strategies were superior to the ‘ASC-US+’ strategy: p16 scored 3+;
HPV16/33/58/31+ &p16+. Moreover, four strategies were favorable to the guideline
strategy: ASC-US+ or p16+; LSIL+ or p16+; HPV16+ or p16+; HSIL+ or p16+ or
HPV16+. These strategies achieved better balance between diseases detection and
colposcopy referral.
CONCLUSIONS: Our findings indicate the promising utility of p16 immunocytology
via adjusting the staining score or serving as an ancillary tool to liquid-based
cytology or combining with genotyping for the triage of HPV-positive
self-samples, which promotes the precise screening of cervical cancer.
Copyright © 2021 Elsevier Inc. All rights reserved.
DOI: 10.1016/j.ygyno.2021.05.014
PMID: 34059349 [Indexed for MEDLINE]
Conflict of interest statement: Declaration of Competing Interest The authors
declare that they have no competing interests.
7. Front Public Health. 2022 Nov 10;10:1010066. doi: 10.3389/fpubh.2022.1010066.
eCollection 2022.
Comparison of cycle threshold values of the Cobas HPV test and viral loads of the BMRT HPV test in cervical cancer screening.
Yang Q(1)(2)(3), Du H(1)(2)(3), Qu X(1)(2), Dai W(1)(2)(3), Gui L(1)(2)(3), Li
C(1)(2)(3), Wang C(1)(2)(3), Guo C(1)(2)(3), Zhang Y(1)(2)(3), Wei L(4),
Belinson JL(5), 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, Peking University-Hong Kong
University of Science and Technology (PKU-HKUST) Medical Center, Shenzhen,
China.
(3)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecologic Diseases, Peking University Shenzhen Hospital, Shenzhen, China.
(4)Department of Obstetrics and Gynecology, Peking University People’s Hospital,
Beijing, China.
(5)Preventive Oncology International, Inc., The Women’s Health Institute,
Cleveland Clinic, Cleveland, OH, United States.
OBJECTIVE: To validate the HPV viral loads that are reflected by the cycle
threshold values of Cobas4800 as the viral load indicators by verifying the
consistency of the viral loads per unit (10,000 cells) from the BMRT assay.
METHODS: The analysis is based on data from the Chinese Multi-Center Screening
Trial (CHIMUST). The cases included in the analysis are all positive for
physician-collected hrHPV on SeqHPV and/or Cobas4800 or negative for hrHPV but
abnormal in cytology (≥LSIL), and some cases selected by nested case-control
randomization from those negative for physician-collected hrHPV and cytology.
With HPV testing results and relevant Ct values from Cobas4800 available, we
tested the entire sample set with the BMRT HPV testing assay and analyzed their
agreement with Cobas4800, followed by a comparison of the CtV from Cobas4800 and
viral loads (lg) from BMRT by lesion grade.
RESULTS: We included 4,485 women (mean age: 45.4 years) in the study, and 4,290
had complete data. The consistency of genotypes from Cobas4800 and BMRT for
hrHPV, HPV-16, HPV-18, and 12-HPV pools was 94.9% (4070/4290, Kappa = 0.827),
99.1% (4251/4290, Kappa = 0.842), 99.6% (4,273/4,290, Kappa = 0.777), and 95.3%
(4,089/4,290, Kappa = 0.821), respectively. Further analysis shows that any
inconsistency between the two assays is likely among samples with comparatively
lower viral loads. When analyzing per lesions of CIN2+ and CIN3+, the CtV from
Cobas4800 and VL (lg) from BMRT are highly correlated inversely and follow the
linear regression for HPV16 and 12-HPV pool (Pearson’s or Spearman’s correlation
coefficient (r): In CIN3+, r HPV16 = -0.641, P < 0.001; r 12-HPVpool = -0.343, P
= 0.109; In CIN2+, r HPV16 = -0.754, P < 0.001; r 12-HPVpool = -0.429, P <
0.001).
CONCLUSION: The CtV from Cobas4800 and the viral loads (lg) of per unit cells
from the BMRT are well correlated for lesion grading when tested on
physician-collected samples. Cobas-CtV is worthy of further study for clinical
application.
Copyright © 2022 Yang, Du, Qu, Dai, Gui, Li, Wang, Guo, Zhang, Wei, Belinson and
Wu.
DOI: 10.3389/fpubh.2022.1010066
PMCID: PMC9686283
PMID: 36438219 [Indexed for MEDLINE]
Conflict of interest statement: Author JB was employed by Preventive Oncology
International, Inc. The remaining authors declare that the research was
conducted in the absence of any commercial or financial relationships that could
be construed as a potential conflict of interest.
8. BMC Womens Health. 2023 Dec 1;23(1):641. doi: 10.1186/s12905-023-02733-1.
Application an internet facilitation in a community-based cervical cancer screening project.
Du H(#)(1)(2)(3), Qu X(#)(2)(3), Wang G(1), Guo C(1), Wang Z(4), Min J(4), Liu
Z(1)(2)(3), Hu Q(1)(2)(3), Luo H(1), Wang C(1)(2)(3), Huang X(1)(2)(3), Chen
Y(1)(5), Wu B(1)(5), Belinson JL(6)(7), Wu R(8)(9)(10).
Author information:
(1)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
No. 1120, Lianhua Road, Shenzhen, 518036, PR China.
(2)Institute of Obstetrics and Gynecology, Shenzhen PKU-HKUST Medical Center,
Shenzhen, PR China.
(3)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecologic Diseases, Shenzhen, PR China.
(4)Pinshan Renmin Hospital, Shenzhen, PR China.
(5)Shenzhen Medical Women’s Association, Shenzhen, PR China.
(6)Preventive Oncology International, Inc, Cleveland Heights, OH, USA.
(7)Women Health Institute, Cleveland Clinic, Cleveland, OH, USA.
(8)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
No. 1120, Lianhua Road, Shenzhen, 518036, PR China. wurf100@126.com.
(9)Institute of Obstetrics and Gynecology, Shenzhen PKU-HKUST Medical Center,
Shenzhen, PR China. wurf100@126.com.
(10)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecologic Diseases, Shenzhen, PR China. wurf100@126.com.
(#)Contributed equally
OBJECTIVE: To evaluate the feasibility of an internet-facilitated community
model for cervical cancer screening using self-collected HPV testing as primary
screening.
METHOD: A population-based cervical cancer screening program was conducted in
the suburb of Shenzhen, China, from September 2014 to July 2017. Women with
25-60 years of age and no pregnancy were eligible for participation.
Participants could register for screening by logging in a website by themselves
or with the aids of local community workers. A unique barcode was issued to each
applicant upon successful registration. After registration, women could get
sampling kits from community screening site/study clinic, collect vaginal
samples privately or in group, and provide their sample for Hr-HPV tests on
Cobas4800 and SeqHPV assays. Testing reports were checkable through personal
account for all participant and phone calls were given to all women positive of
Hr-HPV. Participants positive of both or either the 2 assays were identified as
the positives. The positives could return the study clinic for triage or search
medical care in other clinics. Colposcopy directed or ramdom biopsies were
performed on all positives who returned to the study clinics.
RESULTS: A total of 10,792 community women registered for screening, among whom,
10,010 provided their vaginal samples for tests. 99.5% of the participants were
confirmed to have correct personal identifiable information and samples, and
98.9% of them got HPV testing results from both or either assays. No adverse
event was reported.
CONCLUSION: When self-collected HPV testing is used as the primary testing, the
internet-based data platform facilitates the screening in registration, data
collection, and data tracking, and increases the screening coverage.
Internet-facilitated community model is promising to cervical cancer control and
applicable in regions with variety of resources.
© 2023. The Author(s).
DOI: 10.1186/s12905-023-02733-1
PMCID: PMC10690986
PMID: 38041116 [Indexed for MEDLINE]
Conflict of interest statement: The authors declare no competing interests.
9. 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
reprints. All other permissions can be obtained through our RightsLink service
via the Permissions link on the article page on our site—for further information
please contact journals.permissions@oup.com.
DOI: 10.1093/ajcp/aqad181
PMID: 38365314 [Indexed for MEDLINE]