Use of Multiplex Polymerase Chain Reaction for Detection of High-Risk Human Papillomavirus Genotypes in Women Attending Routine Cervical Cancer Screening in Harare
Background/Aims: In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by...
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Published in | Intervirology Vol. 62; no. 2; pp. 90 - 95 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Basel, Switzerland
01.01.2019
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Subjects | |
Online Access | Get full text |
ISSN | 0300-5526 1423-0100 1423-0100 |
DOI | 10.1159/000502206 |
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Abstract | Background/Aims: In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening. Methodology: Stored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA). Results: A total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR). Conclusion: Multiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare. |
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AbstractList | In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening.
Stored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA).
A total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in -HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR).
Multiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare. In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening.BACKGROUND/AIMSIn Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening.Stored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA).METHODOLOGYStored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA).A total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in -HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR).RESULTSA total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in -HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR).Multiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare.CONCLUSIONMultiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare. Background/Aims: In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening. Methodology: Stored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA). Results: A total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR). Conclusion: Multiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare. |
Author | Dube Mandishora, Racheal Marembo, Takudzwa Borok, Margaret |
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Keywords | Women Zimbabwe Human papillomavirus Multiplex polymerase chain reaction Genotypes |
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References | Cancer Profile In Zimbabwe [Internet]. [cited 2019 Mar 12]. Available from: http://www.zimcancerregistry.co.zw/cancer-profile-in-zimbabwe.html. Rousseau M, Pereira JS, Prado JC. Cervical co-infection with human papillomavirus (HPV) as a predictor of acquisition and persistence of other HPV types. J Infect Dis. 2001;184:1508–17. 10.1086/324579117407250022-1899 Matuvhunye T, Dube-Mandishora RS, Chin’ombe N, Chakafana G, Mbanga J, Zumbika E, et al.. Genotyping Human Papillomavirus in Women Attending Cervical Cancer Screening Clinic in Harare, Zimbabwe. Br Microbiol Res J. 2016Aug;16(6):1–9. 10.9734/BMRJ/2016/284812231-0886 Herald T. First Lady launches HPV vaccination programme [Internet]. The Herald. [cited 2018 Jul 7]. Available from: https://www.herald.co.zw/first-lady-launches-hpv-vaccination-programme Chin’ombe N, Sebata NL, Ruhanya V, Matarira HT. Human papillomavirus genotypes in cervical cancer and vaccination challenges in Zimbabwe. Infect Agent Cancer. 2014May;9(1):16. 10.1186/1750-9378-9-16248473771750-9378 Multiplex PCR Guidelines for Multiplex PCR 5X Master Mix | NEB [Internet]. [cited 2019 Mar 12]. Available from: https://international.neb.com/protocols/2012/09/13/multiplex-pcr-guidelines-for-multiplex-pcr-5x-master-mix-m0284 Nilyanimit P, Chansaenroj J, Poomipak W, Praianantathavorn K, Payungporn S, Poovorawan Y. Comparison of Four Human Papillomavirus Genotyping Methods: Next-generation Sequencing, INNO-LiPA, Electrochemical DNA Chip, and Nested-PCR. Ann Lab Med. 2018Mar;38(2):139–46. 10.3343/alm.2018.38.2.139292147582234-3806 Chirara M, Stanczuk GA, Tswana SA, Nystrom L, Bergstrom S, Moyo SR, et al.. Low risk and high risk human papillomaviruses (HPVs) and cervical cancer in Zimbabwe: epidemiological evidence. Cent Afr J Med. 2001Feb;47(2):32–5. 10.4314/cajm.v47i2.8589119572680008-9176 Bruni L, de Sanjosé S, Barrionuevo-Rosas L, Serrano B. Mena, Bosch FX, et al. Human Papillomavirus and Related Diseases in Zimbabwe. Summary Report. ICO/IARC Information Centre on HPV and Cancer. HPV Information Centre; 2017. Konopnicki D, De Wit S, Clumeck N. HPV and HIV coinfection: a complex interaction resulting in epidemiological, clinical and therapeutic implications. Future Virol. 2013Aug;8(9):903–15. 10.2217/fvl.13.691746-0794 Strickler HD, Burk RD, Fazzari M, Anastos K, Minkoff H, Massad LS, et al.. Natural history and possible reactivation of human papillomavirus in human immunodeficiency virus-positive women. J Natl Cancer Inst. 2005Apr;97(8):577–86. 10.1093/jnci/dji073158408800027-8874 Denny L. Control of cancer of the cervix in low- and middle-income countries. Ann Surg Oncol. 2015Mar;22(3):728–33. 10.1245/s10434-014-4344-8256055131068-9265 World Health Organization. African coalition on maternal, newborn and child health. Africa cervical cancer multi indicator incidence and mortality scorecard. 2014. Cancer Profile In Zimbabwe [Internet]. [cited 2019 Mar 12]. Available from: http://www.zimcancerregistry.co.zw/cancer-profile-in-zimbabwe.html Moscicki AB, Ellenberg JH, Farhat S, Xu J. Persistence of human papillomavirus infection in HIV-infected and -uninfected adolescent girls: risk factors and differences, by phylogenetic type. J Infect Dis. 2004Jul;190(1):37–45. 10.1086/421467151952410022-1899 Romero-Pastrana F. Detection and typing of human papilloma virus by multiplex PCR with type-specific primers. ISRN Microbiol. 2012Mar;2012:186915. 10.5402/2012/186915237243182090-7478 Kuguyo O, Matimba A, Tsikai N, Magwali T, Madziyire M, Gidiri M, et al.. Cervical cancer in Zimbabwe: a situation analysis[Internet]. Pan Afr Med J. 2017Jul;27:215. [cited 2017 Dec 25] Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5622829/10.11604/pamj.2017.27.215.12994289796171937-8688 Schim van der Loeff MF, Mooij SH, Richel O, de Vries HJ, Prins JM. HPV and anal cancer in HIV-infected individuals: a review. Curr HIV/AIDS Rep. 2014Sep;11(3):250–62. 10.1007/s11904-014-0224-x249908101548-3568 Bello BD, Spinillo A, Alberizzi P, Cesari S, Gardella B, D’Ambrosio G, et al.. Cervical infections by multiple human papillomavirus (HPV) genotypes: prevalence and impact on the risk of precancerous epithelial lesions. J Med Virol. 2009Apr;81(4):703–12. 10.1002/jmv.21429192358470146-6615 Dube Mandishora RS, Christiansen IK, Chin’ombe N, Duri K, Ngara B, Rounge TB, et al.. Genotypic diversity of anogenital human papillomavirus in women attending cervical cancer screening in Harare, Zimbabwe. J Med Virol. 2017Sep;89(9):1671–7. 10.1002/jmv.24825283901420146-6615 WHO. | WHO leads the way towards the elimination of cervical cancer as a public health concern [Internet]. WHO. [cited 2019 Jun 1]. Available from: http://www.who.int/reproductivehealth/cervical-cancer-public-health-concern/en ref13 ref12 ref14 ref11 ref10 ref2 ref1 ref8 ref7 ref9 ref4 ref3 ref6 ref5 |
References_xml | – reference: Cancer Profile In Zimbabwe [Internet]. [cited 2019 Mar 12]. Available from: http://www.zimcancerregistry.co.zw/cancer-profile-in-zimbabwe.html. – reference: Bruni L, de Sanjosé S, Barrionuevo-Rosas L, Serrano B. Mena, Bosch FX, et al. Human Papillomavirus and Related Diseases in Zimbabwe. Summary Report. ICO/IARC Information Centre on HPV and Cancer. HPV Information Centre; 2017. – reference: WHO. | WHO leads the way towards the elimination of cervical cancer as a public health concern [Internet]. WHO. [cited 2019 Jun 1]. Available from: http://www.who.int/reproductivehealth/cervical-cancer-public-health-concern/en/ – reference: Denny L. Control of cancer of the cervix in low- and middle-income countries. Ann Surg Oncol. 2015Mar;22(3):728–33. 10.1245/s10434-014-4344-8256055131068-9265 – reference: Strickler HD, Burk RD, Fazzari M, Anastos K, Minkoff H, Massad LS, et al.. Natural history and possible reactivation of human papillomavirus in human immunodeficiency virus-positive women. J Natl Cancer Inst. 2005Apr;97(8):577–86. 10.1093/jnci/dji073158408800027-8874 – reference: Dube Mandishora RS, Christiansen IK, Chin’ombe N, Duri K, Ngara B, Rounge TB, et al.. Genotypic diversity of anogenital human papillomavirus in women attending cervical cancer screening in Harare, Zimbabwe. J Med Virol. 2017Sep;89(9):1671–7. 10.1002/jmv.24825283901420146-6615 – reference: Romero-Pastrana F. Detection and typing of human papilloma virus by multiplex PCR with type-specific primers. ISRN Microbiol. 2012Mar;2012:186915. 10.5402/2012/186915237243182090-7478 – reference: Herald T. First Lady launches HPV vaccination programme [Internet]. The Herald. [cited 2018 Jul 7]. Available from: https://www.herald.co.zw/first-lady-launches-hpv-vaccination-programme/ – reference: Chirara M, Stanczuk GA, Tswana SA, Nystrom L, Bergstrom S, Moyo SR, et al.. 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Snippet | Background/Aims: In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness... In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods... |
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SubjectTerms | Adolescent Adult Aged Aged, 80 and over Cervix Uteri - virology Cross-Sectional Studies DNA, Viral - genetics Early Detection of Cancer - methods Female Genotype HIV Infections - virology Human papillomavirus 16 - genetics Humans Mass Screening Middle Aged Multiplex Polymerase Chain Reaction Papillomaviridae - genetics Papillomavirus Infections - diagnosis Papillomavirus Infections - prevention & control Papillomavirus Infections - virology Research Article Risk Factors Uterine Cervical Neoplasms - diagnosis Uterine Cervical Neoplasms - prevention & control Uterine Cervical Neoplasms - virology Young Adult Zimbabwe |
Title | Use of Multiplex Polymerase Chain Reaction for Detection of High-Risk Human Papillomavirus Genotypes in Women Attending Routine Cervical Cancer Screening in Harare |
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