CHIMUST | Preventive Oncology International, Inc

1. 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.

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.

2. Front Public Health. 2022 May 25;10:840879. doi: 10.3389/fpubh.2022.840879.
eCollection 2022.

Prevalence of Human Papillomavirus Among Chinese Han and Mongols Minority Women in Inner Mongolia, China: Reflected by Self-Collected Samples in CHIMUST.

Guo C(1)(2)(3), Du H(1)(2)(3), Qu X(1)(2), Duan X(4), Li J(5), Li R(1)(2)(3),
Jin H(6), Wang C(1)(2)(3), Zhao C(5), Bao J(1)(2)(3), Luo H(5), Wei L(5),
Belinson JL(7), 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 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)Capital Medical University Beijing Tongren Hospital, Beijing, China.
(5)Peking University People’s Hospital, Beijing, China.
(6)Wushenqi People’s Hospital, Inner Mongolia, China.
(7)Preventive Oncology International, Inc., and the Cleveland Clinic, Cleveland,
OH, United States.

BACKGROUND: The disparities of hr-HPV infection among races/ethnicities have not
been fully discussed. This study aimed to investigate the difference of hr-HPV
infection between Chinese Han and Mongols minority women in Inner Mongolia.
METHODS: Genotyping and histopathology data of Chinese Han and Mongols minority
women in Inner Mongolia from Chinese Multi-Center Screening Trial were used to
analyze the hr-HPV prevalence, and type-specific distribution in abnormal
pathology results.
RESULTS: The hr-HPV infection rates of Han women was 15.9% while of Mongols was
21.6% (P < 0.001). The most prevalent genotypes in Han women were ranked as
HPV-16,-52,-18/-58,-31/-39, and-59 while in Mongols were-16,-31,-58,-18 and-52.
When analyzing the age-specific of hr-HPV infection, two peaks were found at age
of 40-44 (20.5%) and 55-59 (23.5%) years in Han women while three peaks were
observed at age of 30-34 (22.1%), 45-49 (22.9%), and 55-59 (31.8%) years,
respectively, in Mongols. HPV-16 accounting for 62.5 and 53.8% of the CINII+ in
Han and Mongols, respectively.
CONCLUSION: The prevalence of hr-HPV was significantly different between the Han
and Mongols minority women in Inner Mongolia, races/ethnicities background
should be taken into consideration for the refinement of cervical cancer
screening strategies and vaccine implementation in China.

DOI: 10.3389/fpubh.2022.840879
PMCID: PMC9174663
PMID: 35692337 [Indexed for MEDLINE]

Conflict of interest statement: JB was employed by Preventive Oncology
International, Inc., and the Cleveland Clinic. 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.

3. J Clin Epidemiol. 2021 Nov;139:319-329. doi: 10.1016/j.jclinepi.2021.06.009.
Epub 2021 Jun 20.

The prevalence of HR-HPV infection based on self-sampling among women in China exhibited some unique epidemiologic features.

Du H(1), Luo H(2), Wang C(1), Qu X(3), Belinson JL(4), Wu R(5).

Author information:
(1)Department of Gynecology and Obstetrics, Peking University Shenzhen Hospital,
Shenzhen, China; Shenzhen Key Laboratory on Technology for Early Diagnosis of
Major Gynecological Diseases, Shenzhen, PR, China; Institute of Obstetrics and
Gynecology, Shenzhen PKU-HKUST Medical Center, Shenzhen,China.
(2)Department of Gynecology and Obstetrics, Peking University People’ Hospital,
Beijing, PR China.
(3)Institute of Obstetrics and Gynecology, Shenzhen PKU-HKUST Medical Center,
Shenzhen,China.
(4)Cleveland Clinic Lerner College of Medicine, Women’ s Health Institute,
Cleveland, Ohio; Preventive Oncology International, Shaker Heights, Ohio.
(5)Department of Gynecology and Obstetrics, Peking University Shenzhen Hospital,
Shenzhen, China; Shenzhen Key Laboratory on Technology for Early Diagnosis of
Major Gynecological Diseases, Shenzhen, PR, China; Institute of Obstetrics and
Gynecology, Shenzhen PKU-HKUST Medical Center, Shenzhen,China. Electronic
address: wurfpush@126.com.

Objective To investigate the epidemiological characteristics of high-risk human
papillomavirus(HR-HPV) infection based on vaginal self-collected samples. Study
Design and Setting The pooled data of 3045 self-collected samples used for the
analysis derived from four previous studies on cervical cancer screening(The
Chinese Multi-Center Screening Trial, CHIMUST; The Shenzhen Cervical Cancer
Screening Trial-2, SHENCCAST-2; The Chinese Cervical Cancer Prevention Study,
CHIPCAPS; Pingshan trial, PINGSHAN)conducted across China by our team since
2011. These cases were evaluated for HR-HPV type prevalence relative to lesion
grade and age. The occurrence of cervical intraepithelial neoplasia(CIN) with
specific HPV types and the influence of co-infection is explored. Results The
top three most common genotypes among the HR-HPV positives were HPV-52(23.4%),
HPV-16(18.0%), and HPV-58(15.50%). For women with CIN2+, the most frequent
genotypes were HPV-16, 58, 52, and 18 in sequence. HPV-16 accounted for the
majority of CIN2/CIN3/Ca with attribution rate of 23.86%, 44.78% and 50.00%
respectively. HPV-58 accounted for 19.48%, 16.79% and 13.46% respectively. CIN2+
was found in the following types most frequently: HPV-16(31.23%),
HPV-33(24.03%), HPV-58(18.41%), HPV-31(11.76%), HPV-18(7.75%), and
HPV-52(7.30%). HPV-16 showed preference for co-infection with HPV-52 and HPV-58.
Conclusion The prevalence of HR-HPV infection based on self-sampling among women
in China exhibited some unique epidemiologic features.

DOI: 10.1016/j.jclinepi.2021.06.009
PMID: 34161804 [Indexed for MEDLINE]

4. J Med Screen. 2021 Sep;28(3):318-324. doi: 10.1177/0969141320943634. Epub 2020
Sep 1.

The effectiveness of human papillomavirus load, reflected by cycle threshold values, for the triage of HPV-positive self-samples in cervical cancer screening.

Song F(1)(2), Du H(1)(2), Wang C(1)(2), Huang X(1)(2), Qu X(3), Wei L(4),
Belinson JL(3), Wu R(1)(2); CHIMUST team.

Author information:
(1)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
Shenzhen, Guangdong, PR China.
(2)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecological Diseases, Shenzhen, Guangdong, PR China.
(3)Sanming Project of Medicine in Shenzhen Peking University Shenzhen Hospital,
Shenzhen, Guangdong, PR China.
(4)Department of Obstetrics and Gynecology, Peking University People’s Hospital,
Beijing, PR China.

OBJECTIVE: The performance of Cobas4800 cycle threshold value (Ct-value,
reflecting viral load) combined with human papillomavirus (HPV) 16/18 genotyping
was explored as a method of risk stratification to triage patients after primary
HPV screening of self-collected samples.
METHODS: The Chinese Multi-site Screening Trial database was reviewed, with
focus on self-collected samples, using the results of Cobas4800 HPV assay.
Quartiles of Ct-values of each genotype were used for grouping and developing
screening algorithms. Diagnostic accuracy for paired comparisons between
algorithms was obtained using McNemar’s test.
RESULTS: A total of 10,498 women were included. The Ct-values of HPV16 and other
high-risk HPV were inversely correlated with the severity of cervical lesions
(p < 0.001). Risks for cervical intraepithelial neoplasia (CIN2+/CIN3+) were
significantly stratified by Ct-values from channels detecting HPV16 and other
high-risk HPV types. “HPV with HPV16/18 and reflex Ct <33.7” (algorithm G)
achieved a favorable sensitivity to “HPV with atypical squamous cells of
undetermined significance or worse (≥ASCUS)” (81.9% vs. 70.1% for CIN2+,
p < 0.001), a comparable sensitivity to “HPV with HPV16/18 reflex cytology
≥ASCUS” (81.9% vs. 81.3% for CIN2+, p > 0.05), and resulted in a slightly lower
specificity than the latter two algorithms (92.6% vs. 97.4% and 95.4%
respectively for CIN2+, p < 0.05). However, algorithm G achieved a comparable
sensitivity to HPV testing alone for CIN3+, and reduced the colposcopy referral
rate from 13.7% for HPV testing alone to 8.4%.
CONCLUSIONS: HPV viral loads reflected by Ct-values are associated with the
severity of cervical lesions. Ct-values with an appropriate cut-off of 33.7,
combined with HPV16/18 genotyping, represent a promising triage of HPV-positive
women particularly for self-collected samples.

DOI: 10.1177/0969141320943634
PMID: 32869705 [Indexed for MEDLINE]

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+.

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. Infect Agent Cancer. 2021 Mar 25;16(1):21. doi: 10.1186/s13027-021-00360-9.

The prevalence and distribution of human papillomavirus among 10,867 Chinese Han women.

Guo C(#)(1)(2), Du H(#)(1)(2), Belinson JL(3), Wang C(1)(2), Huang X(1)(2), Qu
X(4), Wu R(5)(6); CHIMUST team.

Author information:
(1)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
No. 1120, Lianhua Road, Shenzhen, Guangdong, 518036, PR China.
(2)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecological Diseases, Shenzhen, Guangdong, PR China.
(3)Preventive Oncology International, Inc. Shaker Heights, USA and Cleveland
Clinic, Women’s Health Institute, Cleveland, OH, USA.
(4)Sanming Project of Medicine in Shenzhen Peking University Shenzhen Hospital,
Shenzhen, Guangdong, PR China.
(5)Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital,
No. 1120, Lianhua Road, Shenzhen, Guangdong, 518036, PR China. wurf100@126.com.
(6)Shenzhen Key Laboratory on Technology for Early Diagnosis of Major
Gynecological Diseases, Shenzhen, Guangdong, PR China. wurf100@126.com.
(#)Contributed equally

OBJECTIVE: To assess the prevalence and distribution of HPV genotypes among
Chinese Han women, and to explore the risk of high-grade cervical lesions
associated with individual hr-HPV genotypes.
METHODS: Genotyping and histopathology data from the Chinese Multi-Center
Screening Trial (CHIMUST) and its pilot screening trial, from 6 regions across
mainland China, were re-analyzed. The data from physician- and self-collected
samples from 10,867 Chinese Han women (ages 30-69) were used to determine the
prevalence and distribution of hr-HPV and to explore the risk association
between hr-HPV genotypes and precancerous lesions.
RESULTS: 9.2% of the study population tested hr-HPV positive in
physician-collected sample. The prevalence varied regionally from the lowest in
Guangdong (6.3%) to the highest in Inner Mongolia (13.0%). The most prevalent
genotypes found were HPV-52 (21.7%), HPV-16 (19.2%), HPV-58 (15.0%), HPV-39
(8.9%), and HPV-51 (8.2%). The overall odds ratios for CIN2+ and CIN3+ for the
presence of HPV-16 was 58.6 (95% CI 39.2-87.5) and, 91.6 (95%CI 54.3-154.6),
respectively and remained the highest odds ratio for CIN3+ in all 6 regions.
CONCLUSION: Geographical variation exists in the prevalence and distribution of
hr-HPV in mainland China. HPV-16/52/58 were the most prevalent genotypes, and
HPV-16 had the highest risk for high-grade cervical lesions.
TRIAL REGISTRATION: CHIMUST, Registration number: ChiCTR-EOC-16008456 .
Registered 11 May 2016.

DOI: 10.1186/s13027-021-00360-9
PMCID: PMC7993460
PMID: 33766103

Conflict of interest statement: The authors declare no conflict of interest.

7. 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]

8. 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.

9. 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.

DOI: 10.1186/s13027-020-00328-1
PMCID: PMC7583687
PMID: 33110442

Conflict of interest statement: The authors declare no conflict of interest.

10. PLoS One. 2020 May 7;15(5):e0232107. doi: 10.1371/journal.pone.0232107.
eCollection 2020.

The effectiveness of HPV viral load, reflected by Cobas 4800 HPV-Ct values for the triage of HPV-positive women in primary cervical cancer screening: Direct endocervical samples.

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, China.
(3)Sanming Project of Medicine in Shenzhen Peking University Shenzhen Hospital,
Shenzhen, China.
(4)The Second Hospital of Hebei Medical University, Hebie, China.
(5)Fudan University, Huashan Hospital, Shanghai, China.
(6)Wuhan University, Zhongnan Hospital, Wuhan, China.
(7)Capital Medical University Beijing Tongren Hospital, Beijing, China.
(8)Peking University People’s Hospital, Beijing, China.
(9)Preventive Oncology International, Cleveland Heights, Ohio, United States of
America.
(10)Women’s Health Institute, Cleveland Clinic, Cleveland, Ohio, United States
of America.

OBJECTIVE: To explore the relationship between the viral load reflected by the
Ct value of Cobas 4800 HPV test and cervical lesions, and the effectiveness of
the viral load for secondary triage of HPV-positive women.
METHODS: The Chinese Multi-Center Screening Trial (CHIMUST) evaluated both
self-collected samples and physician-collected samples from women, aged 30 to
59, who were screened for cervical cancer in 6 regions across China. Using
physician collected samples, the relationship between the HPV-Ct values of
different subtypes and the cervical lesions was analyzed. Then the combined use
of the HPV-Ct values with the HPV subtypes was evaluated as a secondary
screening algorithm for the women who were HPV positive.
RESULTS: The Ct values of HPV16 and 12 other HPV subtypes(12-type pool), tested
with Cobas decreased with the progression of cervical lesion (HPV16: r = -0.429,
P<0.001; 12 other HR-HPV subtypes: r = -0.099, P<0.01). The HPV18-Ct value was
not correlated with cervical lesion(P>0.05). Compared with HPV16/18 and cytology
(HPV16/18 positive and 12-type pool plus cytology ≥ ASC-US), the sequential
secondary screening using HPV16/18 and the viral load of 12-type pool (cut-point
HPV-Ct≤31) had equal sensitivities for CIN2+ and CIN3+
(83.1%vs.80.3%,100%vs.92.6%,P>0.05), with slightly lower specificities
(96.2%vs.94.4%,96.5%vs.93.9%,P<0.001) and higher colposcopy referral rate
(4.90%vs.6.59%, P<0.05), but required no cytology.
CONCLUSION: Type-specific HPV viral load is closely related to cervical lesions
severity. It is feasible and efficient to use HPV16/18 and the viral load of 12
other HPV subtypes (with cut-point HPV-Ct≤31) as the secondary screening for HPV
positive women. This algorithm may be useful in low resource regions.

DOI: 10.1371/journal.pone.0232107
PMCID: PMC7205204
PMID: 32379782 [Indexed for MEDLINE]

Conflict of interest statement: Jerome L. Belinson is an expert in the research,
and the entire research has no commercial interest relationship with his POI.
POI did not play a role in the study design, data collection and analysis,
decision to publish, or preparation of the manuscript. This does not alter our
adherence to PLOS ONE policies on sharing data and materials.

11. 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 acervical 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