Next Article in Journal
Demographic and Lifestyle Factors Associated with Patient-Reported Acute COVID-19 Vaccine Reactivity
Previous Article in Journal
COVID-19 Vaccine Acceptance and Hesitancy among Migrants, Refugees, and Foreign Workers: A Systematic Review and Meta-Analysis
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Vaccines
Volume 11
Issue 6
10.3390/vaccines11061071
vaccines-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Clinical Performance of Cobas 6800 for the Detection of High-Risk Human Papillomavirus in Urine Samples

by
Brian Joseph Hajjar
1,
Ummar Raheel
1,*,
Rachel Manina
1,
Jovanie Simpson
1,
Muhammad Irfan
1 and
Yasir Waheed
2,3,*
1
Telostrand Innovations LLC, Hackensack, NJ 07601, USA
2
Office of Research, Innovation & Commercialization, Shaheed Zulfiqar Ali Bhutto Medical University (SZABMU), Islamabad 44000, Pakistan
3
Gilbert and Rose-Marie Chagoury School of Medicine, Lebanese American University, Byblos 1401, Lebanon
*
Authors to whom correspondence should be addressed.
Vaccines 2023, 11(6), 1071; https://doi.org/10.3390/vaccines11061071
Submission received: 23 January 2023 / Revised: 16 April 2023 / Accepted: 8 May 2023 / Published: 6 June 2023
Browse Figure
Review Reports Versions Notes

Abstract

:
Testing for high-risk human papillomavirus (HPV) as part of primary cervical cancer screening has become more common recently. The Cobas 6800, an FDA-approved cervical screening platform, detects 14 high-risk HPVs, including HPV16 and HPV18. However, this test is limited to only women, which leads to low screening rates in trans men and other non-binary people. The cervical screening of trans men and other genders, especially those lying on the female-to-male spectrum, is equally important. Furthermore, cisgender males, particularly homosexuals, are also prone to chronic HPV infections and serve as HPV carriers, transmitting it to women and other men through sexual contact. Another limitation of the test is its invasive specimen collection, which induces discomfort and genital dysphoria. Therefore, there is a need for an innovative, less invasive method that would allow the sampling process to be more comfortable. In this study, we assess the performance of the Cobas 6800 for high-risk HPV detection in urine samples spiked with HPV16, HPV18, and HPV68. The limit of detection (LOD) was calculated using a dilution series (1.25–10,000 copies/mL) over a course of three days. Furthermore, the clinical validation was performed by calculating sensitivity, specificity, and accuracy. The limit of detection ranged from 50–1000 copies/mL depending upon the genotype. Moreover, the urine test demonstrated a high clinical sensitivity of 93%, 94%, and 90% for HPV16, HPV18, and HPV68, with 100% specificity. The overall percent agreement was calculated to be 95% for both HPV16 and HPV18, and 93% for HPV68. The high concordance, reproducibility, and clinical performance of the current assay suggest that the urine-based HPV test fulfills the requirements for its use in primary cervical screening. Moreover, it has the potential to be used for mass screening to not only identify high-risk individuals, but also to monitor vaccine effectiveness.

1. Introduction

Human Papillomavirus (HPV), a member of Papillomaviridae, is a non-enveloped double-stranded DNA virus which is classified into low-risk HPVs involved in causing anogenital and cutaneous warts, and high-risk HPVs (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) that are oncogenic and whose persistent infections are responsible for oropharyngeal and anogenital cancers [1,2,3]. Most HPV infections are asymptomatic, and about 90% of these are cleared by the immune system itself [4,5]. The remaining 10–12% of the infections that are caused by high-risk HPV lead to invasive cancers [6,7].
Cervical cancer is the most common type of HPV-induced cancer worldwide [1], and is the third most common cancer [8], responsible for the highest mortality in women, after breast cancer [9]. About 604,000 HPV-related cases of cervical cancer were reported in 2020 alone, and HPV was found to be responsible for 341,000 cervical-cancer-related deaths [10]. Since most HPV infections do not show any symptoms, routine screening is essential for the prevention and treatment of sexually transmitted infections (STIs). The number of cases of cervical cancer may be reduced by organized screening programs. Moreover, HPV-based screening is more effective than cytology screening in preventing cervical cancer in large randomized controlled studies [11,12].
The Cobas 6800 is an automated high-throughput FDA-approved platform used for routine cervical cancer screening. It uses a quantitative polymerase chain reaction (PCR) to detect 14 types of high-risk HPV along with HPV16 and HPV18. The assay starts with automated nucleic acid extraction from the clinician-obtained samples followed by its binding with magnetic glass particles. The purified nucleic acid is then amplified using a thermostable DNA polymerase. The amplification of sequences of high-risk HPVs and internal control β-globin occurs simultaneously, using specific primers and probes. The automated management software assigns the results for the tests as positive, negative, or invalid [13,14]. The Cobas 6800 is a sensitive HPV detection system, but its utility is limited by the invasive nature of the sampling procedures and the fact that it targets only women. It has been estimated that about 73% of the women attend the screening programs and half of cervical cancer cases are reported among the women who are not willing to attend due to discomfort and embarrassment [15]. Moreover, the female-focused screening misses the fact that both cis and trans men act as carriers for the transmission of HPV to women and other men through sexual intercourse [16]. Furthermore, the rates of cervical cancer screening in transgender men and other non-binary people with an intact cervix is very low compared with cisgender women despite comparable susceptibility to HPV infection [17,18,19,20]. The main reason behind this low rate of screening is the discomfort and genital dysphoria associated with anxiety-provoking pelvic examinations [21,22]. Keeping in view these challenges and limitations, it is necessary to develop and use non-invasive, less painful, and gender-neutral screening approaches, such as urine testing, for detecting HPV infections to circumvent these barriers [23]. The detection of HPV in urine samples is a non-invasive and cost-effective method, which is well accepted by women and trans people in the United States [24,25,26]. The first-void urine testing also has the potential to increase the rates of HPV screening, especially in non-binary people who prefer self-sampling and are not willing to go through an invasive speculum examination [27,28]. In this study, we aim to develop a Cobas 6800-based non-invasive and gender-inclusive test for HPV detection in first-void urine samples.

2. Materials and Methods

2.1. Study Design

The study was performed on a total of 40 urine samples, out of which 10 were negative controls and 30 were spiked positive samples. All the participants, after filling out the informed consent, provided self-collected first-void urine samples which were stored at 2–27 °C. The study was aimed at finding the LOD and evaluating the sensitivity, specificity, reproducibility, and clinical accuracy of the Cobas 6800 for HPV detection using urine samples. All the experiments were carried out at Telostrand Laboratory, LLC, in Hackensack, NJ, USA, a Clinical Laboratory Improvement Amendments (CLIA)-accredited clinical laboratory for high-complexity testing.

2.2. Clinical Samples and HPV Detection Using Cobas 6800

The study included 30 positive urine samples, which were spiked with HPV 16, 18, and 68 virus, and 10 negative samples with no HPV. The positive samples were spiked at different concentrations of the HPV whole organism controls (Exact Diagnostics LLC, Fort Worth, TX, USA) based on LOD data ranging from 1.25 copies/mL to 10,000 copies/mL with a final volume of 1 mL. All the positive spiked and negative urine samples were run on the Cobas 6800 according to the manufacturer’s instructions in triplicate over a period of three days to examine the sensitivity, specificity, percent agreement, and accuracy of the assay. The Cobas 6800 is an automated high-throughput real-time PCR-based system, which is used for routine cervical cancer screening and detects 14 types of high-risk HPV (HPV-31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -66, and -68) along with HPV16 and HPV18. The assay starts with the extraction and release of nucleic acid from the samples, which then binds to the silica surface of the added magnetic glass particles. The purified nucleic acid is then amplified by a PCR reaction using thermostable DNA polymerase. The HPV and β-globin sequences are amplified simultaneously using a universal PCR amplification profile with predefined temperature steps and number of cycles. The automated management software assigns the results for the tests as positive, negative, or invalid [14,15]. In order to assess the sensitivity of the HPV detection test in real urine samples from individuals with a known history of urinary and reproductive health concerns, a pilot study was carried out. The study involved 137 male and 63 female urine samples, and the HPV detection was carried out using similar conditions.

2.3. LOD Studies

The lowest concentration detected in the three replicates was determined as the LOD. The LOD was established with dilution series of 10,000 copies/mL, 5000 copies/mL, 1000 copies/mL, 500 copies/mL, 250 copies/mL, 125 copies/mL, 50 copies/mL, 25 copies/mL, 5 copies/mL, 2.5 copies/mL, and 1.25 copies/mL using whole organism HPV 16, 18, and 68 (Exact Diagnostics LLC) along with internal positive and negative, non-template, and negative extraction controls. All the samples were run thrice, on three different days. The Ct values for assay replicates were averaged, and the values were plotted. The LOD was determined to be between 1.25 and 10,000 copies/mL, depending on the uniformity of the Ct value of the detectable copy number.

2.4. Stability and Reproducibility Analysis

The stability and reproducibility of the assay using urine samples was analyzed with a total of 20 samples, including 5 HPV-negative and 15 HPV-spiked positive samples over a course of seven days. The stability was measured based on the detection of β globulin, and the urine sample was considered valid as long as the β globulin was detectable according to the protocol provided by the manufacturer, Roche Diagnostics. The reproducibility of the assay was assessed by calculating the coefficient of variation (CV%).

2.5. Comparative Analysis of ThinPrep and Urine-Based HPV Detection

The diagnostic accuracy of the Cobas 6800 system for identifying HPV types 16, 18, and 68 in urine was compared with that for identifying ThinPrep samples. First-void urine and ThinPrep samples were obtained from participants for analysis. For both sample types, the sensitivity, specificity, accuracy, and percentage of agreement were determined.

2.6. Statistical Analysis

The clinical sensitivity, specificity, overall percent agreement and accuracy were evaluated for all the samples and calculated as percentages. The Ct values were expressed as mean and standard deviation.

3. Results

3.1. LOD Analysis

Valid test results were obtained for all the positive spiked and negative urine samples using the Cobas 6800. The LOD for HPV16 and HPV18 was established at 50 copies/mL with three out of three positive calls with an average Ct value of 37.03 ± 1.37 and 35.56 ± 0.8, respectively. Similarly, the LOD for HPV68 was measured to be 1000 copies/mL with three out of three positive calls with an average Ct cut-off value of 33.03 ± 0. 31.
No HPV was detected for the negative controls, and the overall concordance was found to be 100% for all the urine samples tested. The LOD results of HPV16, HPV18, and HPV68 plotted against their respective mean Ct values are given in Figure 1.

3.2. Analysis of Clinical Performance

The clinical performance of the Cobas 6800 on urine samples was evaluated by calculating the sensitivity, specificity, accuracy, and overall percent agreement. The validation study was performed over three days for each positive and negative sample to evaluate the clinical performance of the assay. For HPV16, the sensitivity was calculated to be 93% with 100% specificity, as no false positive was recorded. Similarly, the sensitivity for HPV18 was established to be 94% with 100% specificity. For HPV68, the sensitivity was calculated to be 90% and the specificity was 100%, similar to that of HPV16 and HPV18. The overall percent agreement and accuracy for HPV16 and HPV18 were calculated to be 95%. However, both the overall percent agreement and accuracy were 93% for HPV68. The results of the clinical performance of the assay are given in Table 1.
A pilot study on urine samples of patients with a known history of reproductive and urinary health issues was also carried out to assess the sensitivity of the assay for HPV detection in real urine samples. The study included 137 male and 63 female urine samples. The HPV detection was performed on clinical urine samples using similar conditions. The assay showed positive results for HPV in 2 out of 137 male patients and 7 out of 63 female patients. The results of HPV detection in patients’ urine samples are given in Table 2.

3.3. Stability and Reproducibility

The stability study was performed over a course of 7 days on 20 urine samples, of which 5 were negative, and 15 were positive samples spiked with HPV16, HPV18, and HPV68. The samples were stored at 2–8 °C. All 20 samples were run on the Cobas 6800, and the Ct values were recorded and plotted for the complete duration of the stability experiment. For HPV16, there was a shift of approximately four Ct values observed, whereas for HPV18 and HPV68, a shift of two Ct values was observed. The coefficient of variation (CV%) was calculated as between 4.2 and 5.1% for HPV16, 1.9 and 3.6% for HPV18, 2.3 and 3.6% for HPV68, and 1.2 and 4.4% for β globulin, which lies within the acceptable range. For β globulin, this shift was four Ct values. The day 5 stability data for samples 1–5 was lost due to instrument malfunction. The results of stability analyses are given in Table 3.

3.4. Comparison of ThinPrep and Urine-Based HPV Detection

In one of the experimental studies, we compared the diagnostic parameters for detecting HPV16, 18, and 68 in urine and ThinPrep samples (classical/traditional method). It was found that the sensitivity of urine-based HPV detection using the Cobas 6800 for these HPV types ranged from 90% to 94%, whereas the sensitivity for ThinPrep was 100% for all three types. The calculated specificity for both sample types was found to be 100%. Additionally, we evaluated the accuracy and overall percent agreement for urine, which ranged from 93% to 95%, in comparison with 100% for ThinPrep. The results of comparison study are represented in Table 4. Based on the results, we can conclude that both methods, urine-based HPV detection using the Cobas 6800 and the ThinPrep collection method, exhibited comparable sensitivity, specificity, accuracy, and overall percent agreement.

4. Discussion

Screening for cervical cancer has entered a new era, wherein cytology-based examinations are replaced by more sensitive and robust hrHPV DNA testing, whereas cytology is now only performed for triage purposes among women who test positive for HPV. The Cobas 6800 is an automated PCR-based sensitive HPV detection platform using clinician-obtained cervical samples. The test uses DNA amplification and hybridization to detect 14 high-risk HPV genotypes, including HPV16 and HPV18 [29,30]. The sampling process for HPV detection using the Cobas 6800 is often found to be uncomfortable, and women, especially young girls, feel reluctant and embarrassed to go through Pap smears and vaginal examinations [15]. Another major limitation of Cobas HPV testing is that it is gender-specific and mainly targets women. However, HPV screening is equally important for males and non-binary people, especially those with an intact cervix. A screening method that relies on non-invasive self-sampling techniques would not only eliminate the need for a pelvic examination, but it would also eliminate the requirement of a health infrastructure to collect relevant biological samples. The first-void urine-based testing for high-risk HPV detection provides a convenient and less-invasive alternative to the Pap test [27,31]. Several studies have investigated urine-based high-risk HPV detection in an attempt to provide a non-invasive and well-accepted alternative for trans men and other non-binary people who are reluctant to take part in screening programs due to embarrassment and genital dysphoria [24,28,32,33,34]. These factors further support the use of HPV detection in urine as a cervical cancer screening method. In the present study, we optimized and assessed the performance of the Cobas 6800 automated screening platform for HPV detection using spiked urine samples. The Cobas 6800 is an FDA-approved PCR-based HPV detection platform only for clinician-obtained cervical samples, and no study has been performed yet to validate this test for urine samples.
The results of clinical performance showed that the Cobas 6800 performed very well and is compatible with urine samples for HPV detection. The LOD study was performed using urine samples spiked with HPV at a concentration ranging from 10,000–1.25 copies/mL. The findings revealed that the Cobas 6800 can effectively detect both HPV16 and HPV18 at a minimum concentration of 50 copies/mL with an average Ct value of 37 and 35.6, respectively, demonstrating higher sensitivity than the previously recorded results [35]. Moreover, both of the HPV genotypes produced two out of three positive calls at a concentration of 25 copies/mL, which were considered discordant results. These genotypes were completely undetected at much lower copy numbers such as 5, 2.5, and 1.25 copies/mL. The LOD for HPV68, however, was established at 1000 copies/mL with an average Ct value of 33. There were two out of three and one out of three positive calls recorded at lower copy numbers, i.e., 500 and 250 copies/mL, respectively. Previous studies have also shown comparatively less sensitivity and insufficient amplification of HPV68, which can be attributed to the low prevalence and sequence variance in the open reading frame (ORF) of the E6, E7, and L1 genes of HPV68 [36,37]. The internal positive control was also run with all three genotypes and showed high concordance, with an average Ct value of 35. Moreover, no HPV was detected for any of the negative controls. Overall, the Cobas 6800 assay showed good concordance in detecting high-risk HPV in urine samples, which is consistent with HPV detection in clinician-obtained cervical samples [38].
The clinical performance of the assay was performed over a course of three days, and the results showed a high sensitivity, specificity, and accuracy of the urine-based high-risk HPV detection. The overall sensitivity was found to be 93% for HPV16, 94% for HPV18, and 90% for HPV68. The Cobas detected all three high-risk HPVs in spiked urine samples with 100% specificity with no false positive sample detected. Both the overall percent agreement and accuracy were calculated to be 95% for HPV16 and for HPV18, and 93% for HPV68. The sensitivity and specificity of this assay was comparable with HPV detection using clinician-obtained cervical samples [13,29,38]. Moreover, the results were also consistent with the sensitivity and specificity thresholds that are considered adequate for cervical screening [39]. To evaluate the diagnostic performance of HPV detection in urine samples of patients, a pilot study of 137 patients was performed. The assay effectively detected HPV in the first-void urine samples of 2 out of 110 males and 7 out of 63 females. The results of this pilot study, which was conducted using real clinical samples, indicate that the assay is compatible with the detection of HPV in urine samples and has the potential to be used for mass screening.
The stability and reproducibility of urine samples was also assessed in this study over a course of 7 days. The results showed a shift of approximately four Ct values observed over 7 days for HPV16, and β globulin, whereas a shift of only two Ct values was observed for HPV18 and HPV68, indicating a high stability of the urine samples. The reproducibility of the assay was assessed by calculating the coefficient of variation (CV%), which was found to be between 4.2 and 5.1% for HPV16, 1.9 and 3.6% for HPV18, 2.3 and 3.6% for HPV68, and 1.2 and 4.4% for β globulin, which lies within the acceptable range. The Cobas 6800 system also demonstrated high intra-laboratory reproducibility with urine samples when the assay was performed by two different technicians, which proves that the assay is compatible with first-void urine samples for detecting high-risk HPVs. The strength of this study was that the participants included both males and females. A significant amount of research shows that both cis and trans men are the main carriers for HPV and transfer the infection to women and other men through sexual contact. Therefore, HPV screening is equally important for men and non-binary people as for cisgender women [40,41].

5. Conclusions

In conclusion, the current study indicates the clinical utility and compatibility of self-collected first-void urine samples for high-risk HPV detection using the Cobas 6800 system. The assay showed high concordance, sensitivity, specificity, and reproducibility, which demonstrates that urine-based high-risk HPV detection using the Cobas 6800 system meets the requirements for usage in population-based cervical screening. Furthermore, this private, gender-inclusive, and non-invasive assay can circumvent the limitations of the Cobas 6800, and has the potential to increase the participation of young women, non-binary people, trans people, and cisgender men in screening programs. Urine-based HPV detection is cost-effective and well accepted, and can be used for mass screening of populations, allowing for the monitoring of vaccine effectiveness. This, in turn, can not only increase the screening coverage, but will also help the public understand more about HPV and the presence it has in our day-to-day lives.

Author Contributions

Conceptualization: B.J.H. and U.R.; Methodology: U.R., R.M. and B.J.H.; Validation: U.R. and R.M.; Analysis: M.I., U.R. and J.S.; Investigation: U.R. and J.S.; Resources: B.J.H. and J.S.; Data Curation: R.M., M.I. and Y.W.; Writing—Original Draft Preparation: M.I., U.R., R.M. and Y.W.; Writing—Review and Editing: J.S., M.I. and Y.W.; Supervision: U.R. and J.S.; Project Administration: U.R.; Funding Acquisition: B.J.H. and J.S. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

The study was conducted in accordance with the declaration of Helsinki and approved by the Review Board of Telostrand Laboratory.

Informed Consent Statement

Informed consent was obtained from all the subjects involved in the study.

Data Availability Statement

Not applicable.

Acknowledgments

The authors acknowledge John H. Hajjar, Sharon Fagliarone Hajjar, Michael Hajjar, Marc Hajjar and Joseph Diehl Mateer for their excellent technical assistance. Furthermore, the authors thank The Nederlander Producing Company of America, Inc. (New York, NY, USA) for their assistance with this project.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. De Martel, C.; Ferlay, J.; Franceschi, S.; Vignat, J.; Bray, F.; Forman, D.; Plummer, M. Global burden of cancers attributable to infections in 2008: A review and synthetic analysis. Lancet Oncol. 2012, 13, 607–615. [Google Scholar] [CrossRef] [PubMed]
  2. De Martel, C.; Plummer, M.; Vignat, J.; Franceschi, S. Worldwide burden of cancer attributable to HPV by site, country and HPV type. Int. J. Cancer 2017, 141, 664–670. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  3. Buchanan, T.R.; Graybill, W.S.; Pierce, J.Y. Morbidity and mortality of vulvar and vaginal cancers: Impact of 2-, 4-, and 9-valent HPV vaccines. Hum. Vaccines Immunother. 2016, 12, 1352–1356. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  4. Malik, S.; Sah, R.; Muhammad, K.; Waheed, Y. Tracking HPV infection, associated cancer development, and recent treatment efforts—A comprehensive review. Vaccines 2023, 11, 102. [Google Scholar] [CrossRef] [PubMed]
  5. Waheed, Y.; Sah, R.; Muhammad, K. Recent developments in vaccines for viral diseases. Vaccines 2023, 11, 198. [Google Scholar] [CrossRef] [PubMed]
  6. Pinto, A.P.; Crum, C.P. Natural history of cervical neoplasia: Defining progression and its consequence. Clin. Obstet. Gynecol. 2000, 43, 352–362. [Google Scholar] [CrossRef]
  7. Myers, E.R.; McCrory, D.C.; Nanda, K.; Bastian, L.; Matchar, D. Mathematical model for the natural history of human papillomavirus infection and cervical carcinogenesis. Am. J. Epidemiology 2000, 151, 1158–1171. [Google Scholar] [CrossRef] [Green Version]
  8. Magalhães, G.M.; Vieira, É.C.; Garcia, L.C.; Carvalho-Leite, D.; de Lourdes Ribeiro, M.; Guedes, A.C.M.; Araújo, M.G. Update on human papilloma virus-part I: Epidemiology, pathogenesis, and clinical spectrum. An. Bras. Dermatol. 2021, 96, 1–16. [Google Scholar] [CrossRef]
  9. Bray, F.; Ferlay, J.; Soerjomataram, I.; Siegel, R.L.; Torre, L.A.; Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 2018, 68, 394–424. [Google Scholar] [CrossRef] [Green Version]
  10. Sung, H.; Ferlay, J.; Siegel, R.L.; Laversanne, M.; Soerjomataram, I.; Jemal, A.; Bray, F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 2021, 71, 209–249. [Google Scholar] [CrossRef]
  11. Dillner, J.; Rebolj, M.; Birembaut, P.; Petry, K.-U.; Szarewski, A.; Munk, C.; De Sanjose, S.; Naucler, P.; Lloveras, B.; Kjaer, S.; et al. Long term predictive values of cytology and human papillomavirus testing in cervical cancer screening: Joint European cohort study. BMJ 2008, 337, a1754. [Google Scholar] [CrossRef] [Green Version]
  12. Ronco, G.; Dillner, J.; Elfström, K.M.; Tunesi, S.; Snijders, P.J.; Arbyn, M.; Kitchener, H.; Segnan, N.; Gilham, C.; Giorgi-Rossi, P.; et al. Efficacy of HPV-based screening for prevention of invasive cervical cancer: Follow-up of four European randomised controlled trials. Lancet 2014, 383, 524–532. [Google Scholar] [CrossRef]
  13. Frayle, H.; Gori, S.; Rizzi, M.; Graziani, B.N.; Vian, E.; Rossi, P.G.; Del Mistro, A. HPV testing for cervical cancer screening: Technical improvement of laboratory logistics and good clinical performance of the cobas 6800 in comparison to the 4800 system. BMC Women’s Health 2019, 19, 47. [Google Scholar] [CrossRef]
  14. Aretzweiler, G.; Leuchter, S.; Simon, C.O.; Marins, E.; Frontzek, A. Generating timely molecular diagnostic test results: Workflow comparison of the cobas® 6800/8800 to Panther. Expert Rev. Mol. Diagn. 2019, 19, 951–957. [Google Scholar] [CrossRef] [Green Version]
  15. Dugué, P.-A.; Lynge, E.; Bjerregaard, B.; Rebolj, M. Non-participation in screening: The case of cervical cancer in Denmark. Prev. Med. 2012, 54, 266–269. [Google Scholar] [CrossRef]
  16. Hernandez, B.Y.; Wilkens, L.R.; Zhu, X.; Thompson, P.; McDuffie, K.; Shvetsov, Y.B.; Kamemoto, L.E.; Killeen, J.; Ning, L.; Goodman, M.T. Transmission of human papillomavirus in heterosexual couples. Emerg. Infect. Dis. 2008, 14, 888. [Google Scholar] [CrossRef]
  17. Espersen, M.M.; Holten, I.W. Barriers in screening for cervical cancer. Ugeskr. Laeger 2005, 167, 4371–4374. [Google Scholar]
  18. Reisner, S.L.; Deutsch, M.B.; Peitzmeier, S.M.; Hughto, J.M.W.; Cavanaugh, T.P.; Pardee, D.J.; McLean, S.A.; Panther, L.A.; Gelman, M.; Mimiaga, M.J.; et al. Test performance and acceptability of self-versus provider-collected swabs for high-risk HPV DNA testing in female-to-male trans masculine patients. PLoS ONE 2018, 13, e0190172. [Google Scholar] [CrossRef] [Green Version]
  19. Harb, C.Y.; Pass, L.E.; De Soriano, I.C.; Zwick, A.; Gilbert, P.A. Motivators and barriers to accessing sexual health care services for transgender/genderqueer individuals assigned female sex at birth. Transgender Heal. 2019, 4, 58–67. [Google Scholar] [CrossRef] [Green Version]
  20. Peitzmeier, S.M.; Reisner, S.L.; Harigopal, P.; Potter, J. Female-to-male patients have high prevalence of unsatisfactory Paps compared to non-transgender females: Implications for cervical cancer screening. J. Gen. Intern. Med. 2014, 29, 778–784. [Google Scholar] [CrossRef] [Green Version]
  21. Potter, J.; Peitzmeier, S.M.; Bernstein, I.; Reisner, S.L.; Alizaga, N.M.; Agénor, M.; Pardee, D.J. Cervical cancer screening for patients on the female-to-male spectrum: A narrative review and guide for clinicians. J. Gen. Intern. Med. 2015, 30, 1857–1864. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  22. Agénor, M.; Peitzmeier, S.M.; Bernstein, I.M.; McDowell, M.; Alizaga, N.M.; Reisner, S.L.; Pardee, D.J.; Potter, J. Perceptions of cervical cancer risk and screening among transmasculine individuals: Patient and provider perspectives. Cult. Heal. Sex. 2016, 18, 1192–1206. [Google Scholar] [CrossRef] [PubMed]
  23. Nilyanimit, P.; Chansaenroj, J.; Karalak, A.; Laowahutanont, P.; Junyangdikul, P.; Poovorawan, Y. Comparison of human papillomavirus (HPV) detection in urine and cervical swab samples using the HPV GenoArray Diagnostic assay. PeerJ 2017, 5, e3910. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  24. D’Hauwers, K.W.; Tjalma, W.A. Screening for human papillomavirus: Is urine useful? Indian J. Cancer 2009, 46, 190. [Google Scholar] [CrossRef]
  25. Cuzick, J.; Cadman, L.; Ahmad, A.S.; Ho, L.; Terry, G.; Kleeman, M.; Lyons, D.; Austin, J.; Stoler, M.H.; Vibat, C.R.T.; et al. Performance and Diagnostic Accuracy of a Urine-Based Human Papillomavirus Assay in a Referral PopulationPredictors 4 Substudy: HPV Testing from Urine. Cancer Epidemiol. Biomark. Prev. 2017, 26, 1053–1059. [Google Scholar] [CrossRef] [Green Version]
  26. Tabaac, A.R.; Sutter, M.E.; Wall, C.S.; Baker, K.E. Gender identity disparities in cancer screening behaviors. Am. J. Prev. Med. 2018, 54, 385–393. [Google Scholar] [CrossRef]
  27. Vorsters, A.; Bergh, J.V.D.; Micalessi, I.; Biesmans, S.; Bogers, J.-P.; Henß, A.; De Coster, I.; Ieven, M.; Van Damme, P. Optimization of HPV DNA detection in urine by improving collection, storage, and extraction. Eur. J. Clin. Microbiol. Infect. Dis. 2014, 33, 2005–2014. [Google Scholar] [CrossRef]
  28. Enerly, E.; Olofsson, C.; Nygård, M. Monitoring human papillomavirus prevalence in urine samples: A review. Clin. Epidemiol. 2013, 5, 67. [Google Scholar] [CrossRef] [Green Version]
  29. Sundström, K.; Lamin, H.; Dillner, J. Validation of the cobas 6800 human papillomavirus test in primary cervical screening. PLoS ONE 2021, 16, e0247291. [Google Scholar] [CrossRef]
  30. Lloveras, B.; Gomez, S.; Alameda, F.; Bellosillo, B.; Mojal, S.; Muset, M.; Parra, M.; Palomares, J.C.; Serrano, S. HPV testing by cobas HPV test in a population from Catalonia. PLoS ONE 2013, 8, e58153. [Google Scholar] [CrossRef]
  31. Pathak, N.; Dodds, J.; Zamora, J.; Khan, K.S. Accuracy of urinary human papillomavirus testing for presence of cervical HPV: Systematic review and meta-analysis. BMJ 2014, 349, g5264. [Google Scholar] [CrossRef] [Green Version]
  32. Burroni, E.; Bonanni, P.; Sani, C.; Lastrucci, V.; Carozzi, F. Human papillomavirus prevalence in paired urine and cervical samples in women invited for cervical cancer screening. J. Med Virol. 2015, 87, 508–515. [Google Scholar] [CrossRef]
  33. Hagihara, M.; Yamagishi, Y.; Izumi, K.; Miyazaki, N.; Suzuki, T.; Kato, H.; Nishiyama, N.; Koizumi, Y.; Suematsu, H.; Mikamo, H. Comparison of initial stream urine samples and cervical samples for detection of human papillomavirus. J. Infect. Chemother. 2016, 22, 559–562. [Google Scholar] [CrossRef]
  34. Sahasrabuddhe, V.V.; Gravitt, P.E.; Dunn, S.T.; Brown, D.; Allen, R.A.; Eby, Y.J.; Smith, K.; Zuna, R.E.; Zhang, R.R.; Gold, M.A.; et al. Comparison of human papillomavirus detections in urine, vulvar, and cervical samples from women attending a colposcopy clinic. J. Clin. Microbiol. 2014, 52, 187–192. [Google Scholar] [CrossRef] [Green Version]
  35. Rao, A.; Young, S.; Erlich, H.; Boyle, S.; Krevolin, M.; Sun, R.; Apple, R.; Behrens, C. Development and characterization of the cobas human papillomavirus test. J. Clin. Microbiol. 2013, 51, 1478–1484. [Google Scholar] [CrossRef] [Green Version]
  36. Eklund, C.; Zhou, T.; Dillner, J. Global proficiency study of human papillomavirus genotyping. J. Clin. Microbiol. 2010, 48, 4147–4155. [Google Scholar] [CrossRef] [Green Version]
  37. Estrade, C.; Sahli, R. Updating the PGMY primers and probes for improved detection of HPV68a: Validation of version 2 of the PGMY-CHUV assay. J. Clin. Microbiol. 2014, 52, 4033–4035. [Google Scholar] [CrossRef] [Green Version]
  38. Saville, M.; Sultana, F.; Malloy, M.J.; Velentzis, L.S.; Caruana, M.; Ip, E.L.O.; Keung, M.H.T.; Canfell, K.; Brotherton, J.M.L.; Hawkes, D. Clinical Validation of the cobas HPV Test on the cobas 6800 System for the Purpose of Cervical Screening. J. Clin. Microbiol. 2019, 57, e01239-18. [Google Scholar] [CrossRef] [Green Version]
  39. Meijer, C.J.; Berkhof, J.; Castle, P.E.; Hesselink, A.T.; Franco, E.L.; Ronco, G.; Arbyn, M.; Bosch, F.X.; Cuzick, J.; Dillner, J.; et al. Guidelines for human papillomavirus DNA test requirements for primary cervical cancer screening in women 30 years and older. Int. J. Cancer 2009, 124, 516–520. [Google Scholar] [CrossRef] [Green Version]
  40. Xavier, S.D.; Castro, T.M.P.P.G.; Filho, I.B.; de Carvalho, J.M.; Framil, V.M.D.S.; Syrjänen, K.J. Prevalence of human papillomavirus (HPV) DNA in oral mucosa of men with anogenital HPV infection. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endodontol. 2009, 108, 732–737. [Google Scholar] [CrossRef]
  41. Giuliano, A.R.; Tortolero-Luna, G.; Ferrer, E.; Burchell, A.N.; de Sanjose, S.; Kjaer, S.K.; Muñoz, N.; Schiffman, M.; Bosch, F.X. Epidemiology of human papillomavirus infection in men, cancers other than cervical and benign conditions. Vaccine 2008, 26 (Suppl. 10), K17–K28. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Vaccines 11 01071 g001 550
Figure 1. LOD for HPV16, HPV18, and HPV68.
Figure 1. LOD for HPV16, HPV18, and HPV68.
Vaccines 11 01071 g001
Table
Table 1. Summary of clinical performance of Cobas 6800.
Table 1. Summary of clinical performance of Cobas 6800.
Total Samples/CallsHPV 16HPV 18HPV 68
12012090
True Positive848554
False Positive000
True Negative 303030
False Negative 656
Estimated Sensitivity =100% × TP/(TP + FN)=100% × TP/(TP + FN)=100% × TP/(TP + FN)
93%94%90%
Estimated Specificity 100% × TN/(FP + TN)100% × TN/(FP + TN)100% × TN/(FP + TN)
100%100%100%
Overall Percent Agreement =100% × (TP + TN)/Total=100% × (TP + TN)/Total=100% × (TP + TN)/Total
95%95%93%
Accuracy 95%95%93%
TP: True Positive, TN: True Negative, FP: False Positive, FN: False Negative.
Table
Table 2. Pilot validation study using clinical urine samples.
Table 2. Pilot validation study using clinical urine samples.
Clinical SamplesPositiveNegativeInvalid
Male11021080
Female637560
Total Samples17391640
Table
Table 3. Detailed table of stability and reproducibility analysis on 20 samples over a period of 7 days. Neg: Negative, IM: Instrument Malfunction.
Table 3. Detailed table of stability and reproducibility analysis on 20 samples over a period of 7 days. Neg: Negative, IM: Instrument Malfunction.
HPV16HPV18
SamplesDay 1Day 2Day 3Day 4Day 5Day 6Day 7Day 1Day 2Day 3Day 4Day 5Day 6Day 7
Sample 130.0931.4932.0532.72IM34.5834.0928.4928.929.3529.78IM30.9931.16
Sample 230.3931.4531.7432.56IM33.9933.5628.8629.2129.4329.64IM29.9330.48
Sample 330.2431.4131.9232.74IM34.8233.6428.6828.7729.3329.76IM30.6630.89
Sample 430.5631.8731.7432.67IM34.733.6328.7229.0729.3129.97IM30.4130.59
Sample 530.4631.4932.0932.74IM34.7933.7328.7729.329.429.77IM30.9530.54
Sample 630.2831.4331.8832.7933.4434.3733.7328.7428.8929.3429.9830.6130.7830.8
Sample 730.3331.131.8432.7133.2635.1333.7928.6628.4629.3529.9430.1530.7130.84
Sample 830.1231.4231.8432.5933.4734.333.7328.829.129.1430.1230.9430.6430.63
Sample 930.2931.2831.6932.4733.0534.8434.128.8728.729.1729.5730.2330.931.12
Sample 1030.7931.3731.6132.5332.9634.8333.4229.1429.1829.129.8630.2131.0130.6
Sample 1130.3231.4431.8332.6933.4334.9933.7328.4928.929.0529.6929.9730.4930.85
Sample 1230.3831.7431.6832.4733.5135.0833.5428.7229.3429.2729.4930.3630.8831.18
Sample 1330.2831.1731.6932.5533.4334.9933.7128.7628.8129.1929.5130.9730.8830.83
Sample 1430.3131.431.6432.3633.3534.8433.9128.6628.929.1829.6830.4730.330.69
Sample 1530.1631.5331.6932.4533.2534.933.6328.5129.129.1929.6230.0930.8730.58
Sample 16NegNegNegNegNegNegNegNegNegNegNegNegNegNeg
Sample 17NegNegNegNegNegNegNegNegNegNegNegNegNegNeg
Sample 18NegNegNegNegNegNegNegNegNegNegNegNegNegNeg
Sample 19NegNegNegNegNegNegNegNegNegNegNegNegNegNeg
Sample 20NegNegNegNegNegNegNegNegNegNegNegNegNegNeg
Average30.3331.4331.832.633.3234.7433.7328.7228.9829.2529.7630.430.6930.78
HPV68β globulin
SamplesDay 1Day 2Day 3Day 4Day 5Day 6Day 7Day 1Day 2Day 3Day 4Day 5Day 6Day 7
Sample 129.9630.4430.9931.47IM32.6332.723031.2131.1331.58N/AN/A32
Sample 230.0830.7630.9831.42IM31.5632.2230.0931.2930.7731.56N/A32.731.41
Sample 329.930.2431.0231.58IM32.6532.4529.7330.9830.7231.33N/A33.832.07
Sample 430.2930.6730.8131.68IM32.5132.2230.0531.5430.7831.46N/A33.2631.85
Sample 530.2930.731.1531.46IM32.7632.3230.6931.1530.7831.1N/A33.9431.66
Sample 630.2130.4631.0131.6432.2832.3132.330.0831.2731.0531.6331.9733.6131.87
Sample 730.0530.1431.1331.6532.0532.7432.1929.6830.6131.3231.2531.5933.8232.27
Sample 829.8830.6830.8131.6332.3632.5532.429.5530.7930.5131.1832.0333.4232.35
Sample 930.0730.2830.9931.3232.0732.7232.729.8830.3230.6231.2431.77N/A32.73
Sample 1030.4430.3530.6331.3931.8932.7232.4630.4629.1830.5831.4131.49N/A31.66
Sample 1130.0330.5231.131.4431.8532.3932.3430.123130.7331.8131.8633.732.54
Sample 1230.2230.830.8831.4132.2732.6532.2230.2931.8430.3931.7532.4733.7731.75
Sample 1330.1430.6430.8731.4732.0432.6432.329.9231.3930.5631.3532.3433.731.86
Sample 1429.9830.3130.5731.2632.332.5732.5229.6431.1330.5531.632.4733.5932.09
Sample 1529.8830.5830.9731.3931.9932.731.6629.3630.8331.1131.5631.48N/A32.16
Sample 16NegNegNegNegNegNegNeg30.7931.8432.0231.5132.131.2731.5
Sample 17NegNegNegNegNegNegNeg31.6531.5231.6531.0932.73132.18
Sample 18NegNegNegNegNegNegNeg31.7731.231.2331.5132.331.8232
Sample 19NegNegNegNegNegNegNeg31.2831.0932.1629.732.1231.7231.72
Sample 20NegNegNegNegNegNegNeg30.9331.8231.2332.4632.0331.7231.3
Average30.0930.530.9231.4832.1132.5432.3329.9630.9730.7731.431.933.5732
Table
Table 4. Comparison of ThinPrep and urine-based HPV detection using Cobas 6800.
Table 4. Comparison of ThinPrep and urine-based HPV detection using Cobas 6800.
HPV16HPV18HPV68
Sample TypeThinPrepUrineThinPrepUrineThinPrepUrine
Total Samples/Calls30120301203090
True Positive308430853054
False Positive000000
True Negative303030303030
False Negative060506
Sensitivity = 100% × TP/(TP + FN)100%93%100%94%100%90%
Specificity = 100% × TN/(FP + TN)100%100%100%100%100%100%
Accuracy = 100% × (TP + TN)/Total100%95%100%95%100%93%
Overall Percent Agreement100%95%100%95%100%93%
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Hajjar, B.J.; Raheel, U.; Manina, R.; Simpson, J.; Irfan, M.; Waheed, Y. Clinical Performance of Cobas 6800 for the Detection of High-Risk Human Papillomavirus in Urine Samples. Vaccines 2023, 11, 1071. https://doi.org/10.3390/vaccines11061071

AMA Style

Hajjar BJ, Raheel U, Manina R, Simpson J, Irfan M, Waheed Y. Clinical Performance of Cobas 6800 for the Detection of High-Risk Human Papillomavirus in Urine Samples. Vaccines. 2023; 11(6):1071. https://doi.org/10.3390/vaccines11061071

Chicago/Turabian Style

Hajjar, Brian Joseph, Ummar Raheel, Rachel Manina, Jovanie Simpson, Muhammad Irfan, and Yasir Waheed. 2023. "Clinical Performance of Cobas 6800 for the Detection of High-Risk Human Papillomavirus in Urine Samples" Vaccines 11, no. 6: 1071. https://doi.org/10.3390/vaccines11061071

APA Style

Hajjar, B. J., Raheel, U., Manina, R., Simpson, J., Irfan, M., & Waheed, Y. (2023). Clinical Performance of Cobas 6800 for the Detection of High-Risk Human Papillomavirus in Urine Samples. Vaccines, 11(6), 1071. https://doi.org/10.3390/vaccines11061071

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop