Prevalence of Human Papillomavirus (HPV) 16 and 18 in oral malignant and potentially malignant disorders: A polymerase chain reaction analysis – A comparative study

Objective Human papillomavirus (HPV) has been identified as the principal etiologic agent for cervical cancer and its precursors. Different HPV types have been associated with different oncogenic potential. The purpose of this study was to evaluate the relationship between specific HPV type infection and expression pattern of the ras family oncogenes in different grades of HPV-associated human cervical neoplasia. Methods HPV typing was performed using polymerase chain reaction (PCR) in 31 HPV-positive human cervical specimens from patients with squamous intraepithelial lesions (SIL) or squamous cervical carcinoma (SCC). The mRNA expression levels of H-, K- and N-ras oncogenes were examined using the reverse transcriptase polymerase chain reaction (RT-PCR) technique. Statistical analyses were performed using SPSS software. Results Among patients with SCC, H-, K- and N-ras expression levels were higher in HPV 16/18-associated cases compared to HPV 16/18-unassociated samples (p=0.003, p=0.004 and p=0.0001, respectively). The expression levels for H-, K-and N-ras were significantly higher in SCC patients with multiple HPV infection compared with SCC patients with single HPV infection (p=0.009, p=0.01 and p=0.021, respectively). Among patients with SIL, no statistically significant relationship was found between ras expression and HPV status. Conclusion Our findings indicate the possible role of ras signaling interaction with “high-risk” HPV 16/18 and multiple HPV infection in cervical cancer development.

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Detection of Human Papillomavirus in Cervical Gradings by Immunohistochemistry and Typing of HPV 16 and 18 in High-Grades by Polymerase Chain Reaction

Journal of Laboratory Physicians ◽

10.4103/0974-2727.66711 ◽

2010 ◽

Vol 2 (01) ◽

pp. 031-036 ◽

Cited By ~ 3

Author(s):

Mrudula Soma ◽

Suhasini Kamaraj

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

High Risk ◽

Tamil Nadu ◽

Low Grade ◽

Chain Reaction ◽

Hpv 16 ◽

Screening Programs ◽

South Indian ◽

Polymerase Chain

ABSTRACT Background: Cervical cancer has a major impact on developing countries, where screening programs are not well established or effective. Aim: This study aims to investigate Human papillomavirus (HPV) 6, 11 and 18 expression in cervical biopsies by immunohistochemistry, (IHC) followed by typing of high-risk HPV 16 and 18 in high-grades by polymerase chain reaction (PCR). Settings and Design: During the study period of six months, 30 biopsy samples were obtained from patients attending various gynecology clinics in and around Trichy District, Tamil Nadu, between January and June 2009. Materials and Methods: The ecto- and endoscopic biopsy specimens of the cervix were fixed in 10% buffered formalin; routine paraffin sections were taken for processing and stained with hematoxylin and eosin. The samples were graded as Normal cervicitis, Cervical intraepithelial neoplasia (CIN) I, II, III, and squamous cell carcinoma (SCC), for original diagnosis by pathologists. The extra sections were studied for the expression of HPV 6, 11 and 18 by immunohistochemistry and HPV DNA 16 and18 by PCR. Results: Out of thirty samples, 15 expressed positive and 15 negative for HPV marker. Twenty-seven cases of cervical gradings have been categorized into high grade CIN II/III, SCC (23) and low grade CIN I (4). The high grades were subjected to PCR for high-risk typing. The results revealed that 15 cases were positive for HPV genotype 16 and eight cases for HPV genotype 18. The prevalence of HPV infection was found to be higher in women aged between 50 and 59. Conclusion: This study reveals a significant detection of HPV in the South Indian suspected individuals, by the use of advanced techniques such as IHC and PCR.

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Human papillomavirus genotyping by a polymerase chain reaction-based genechip method in cervical carcinoma treated with neoadjuvant chemotherapy plus radical surgery

International Journal of Gynecological Cancer ◽

10.1136/ijgc-00009577-200407000-00012 ◽

2004 ◽

Vol 14 (4) ◽

pp. 639-649 ◽

Cited By ~ 4

Author(s):

H. J. Huang ◽

S. L. Huang ◽

C. Y. Lin ◽

R. W. Lin ◽

F. Y. Chao ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

Cervical Carcinoma ◽

Radical Surgery ◽

Chain Reaction ◽

Hpv 16 ◽

Hpv Dna ◽

Hpv Genotyping ◽

Polymerase Chain ◽

Hpv 18

The aim of this study was to evaluate the accuracy of human papillomavirus (HPV) genotyping by a polymerase chain reaction (PCR)-based genechip method and to determine the prognostic value of HPV genotype in bulky stage IB or IIA cervical carcinoma treated with neoadjuvant chemotherapy (NAC) and radical surgery. A total of 149 patients had adequate tissue for the study. The SPF1/GP6+ primers were used to amplify a 184 bp fragment. The amplimers were submitted for direct sequencing and hybridization with a genechip using revert-blot detection of 39 types of HPV DNA in a single reaction. Two runs of PCR with respective hybridization were performed for each tumor. The complete concordance of HPV genotyping was 80.5% (120/149) of the paired genechip results. The kappa coefficient was 0.634 (P < 0.0001). HPV DNA sequences were detected in 100% of the specimens, among which 67.8% harbored single type and 32.2% contained multiple types. HPV-16 was detected in 98.7%, HPV-18 in 22.8%, HPV-31 in 0.7%, HPV-45 in 1.3%, HPV-52 in 2.0%, HPV-58 in 6.7%, HPV-59 in 4.7%, and HPV-67 in 0.7%. In multivariate analyses, the HPV genotype [HPV-18 or HPV-16 and HPV-18 only versus all others: relative risk (RR), 2.33; 95% CI, 1.17–4.64; P = 0.016] and pre-NAC tumor size (>5 versus ≤5 cm: RR, 2.25; 95% CI, 1.13–4.48; P = 0.021) were significantly related to overall survival. This PCR-based genechip method is sensitive and reproducible for HPV genotyping. The association of HPV-18 or HPV-16 and HPV-18 with poor outcome in cervical carcinoma treated with NAC plus radical surgery is confirmed.

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Molecular Methods for a Correct Diagnosis of Multiple HPV Infections and Clinical Implications for Vaccine

International Journal of Gynecological Cancer ◽

10.1097/igc.0b013e31820f5eed ◽

2011 ◽

Vol 21 (3) ◽

pp. 545-550 ◽

Cited By ~ 4

Author(s):

Andrea Tinelli ◽

Giuseppe Leo ◽

Domenico Dell'Edera ◽

Fabio Storelli ◽

Maria Maddalena Galante ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

Real Time ◽

Sequence Homology ◽

Direct Sequencing ◽

Chain Reaction ◽

Hpv 16 ◽

Hpv Genotyping ◽

Polymerase Chain ◽

Hpv 18

IntroductionThe human papillomavirus (HPV) family is characterized by minimal genotypic differences corresponding to different virus types. The aim of this study was to detect the HPV coinfections and the inner genotype in a series of 336 cervical-vaginal samples.MethodsA total of 336 cervical-vaginal samples were taken from 2007 to 2009 using specific molecular techniques such as molecular sequencing and hybridizations. The genome amplification of the L1 open reading frame was analyzed by real-time polymerase chain reaction; direct sequencing was performed by SYBR green fluorescent molecule and degenerate primers MY09 and MY11. The HPV genotyping was accomplished via oligonucleotide probe hybridization. The phylogenetic correlations in coinfections were analyzed by sequence homology of the L1 genomic region. Identified genotypes were then compared.ResultsHuman papillomavirus positivity was observed in 125 cases (37.2%), with 21 cases (16.8%) of HPV presence in coinfections. Coinfections involved HPV 16 genotype (8 cases) and HPV 18 (5 cases). The HPV 16 infection was mainly associated with genotypes with a lower-than-broad sequence homology, so the HPV 18 was linked to genotypes represented in the opposite phylogenetic tree.ConclusionsThe combined and steady use of diagnostic procedures, such as real-time polymerase chain reaction, molecular hybridization, direct sequencing, and HPV genotyping test, allow accurate diagnosis of monoinfections and coinfections. This may faciliate the development of specific viral tests and prophylactic anti-HPV vaccines.

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Comparative study of DNA extraction methodologies from goat sperm and its effects on polymerase chain reaction analysis

Genetics and Molecular Research ◽

10.4238/2014.august.7.21 ◽

2014 ◽

Vol 13 (3) ◽

pp. 6070-6078 ◽

Cited By ~ 9

Author(s):

E.C.B. Silva ◽

M.A. Pelinca ◽

A.C. Acosta ◽

D.M.F. Silva ◽

M.A. Gomes Filho ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Comparative Study ◽

Dna Extraction ◽

Polymerase Chain Reaction Analysis ◽

Chain Reaction ◽

Polymerase Chain

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Quantitative real-time polymerase chain reaction analysis of the type distribution, viral load, and physical status of human papillomavirus in liquid-based cytology samples from cervical lesions

International Journal of Gynecological Cancer ◽

10.1111/j.1525-1438.2007.00979.x ◽

2008 ◽

Vol 18 (1) ◽

pp. 121-127 ◽

Cited By ~ 21

Author(s):

T. YOSHIDA ◽

T. SANO ◽

T. KANUMA ◽

N. OWADA ◽

S. SAKURAI ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

Viral Load ◽

Polymerase Chain Reaction Analysis ◽

Physical Status ◽

Cervical Lesions ◽

Chain Reaction ◽

Type Distribution ◽

Liquid Based Cytology ◽

Polymerase Chain

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Identifikasi Type Human Papillomavirus (HPV) pada Penderita Kanker Serviks

Jurnal Sains Farmasi & Klinis ◽

10.29208/jsfk.2016.3.1.100 ◽

2016 ◽

Vol 3 (1) ◽

pp. 54

Author(s):

Marlina Marlina ◽

Yufri Aldi ◽

Andani Eka Putra ◽

Densi Selpia Sopianti ◽

Dewi Gulyla Hari ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

High Risk ◽

Chain Reaction ◽

Hpv 16 ◽

Polymerase Chain

Human Papillomavirus (HPV) merupakan faktor resiko yang paling signifikan penyebab dari kanker serviks. Tujuan penelitian ini untuk identifikasi tipe HPV 16, 18, 31, 33, 45 dan 52 pada pasien kanker serviks. HPV tipe ini merupakan deretan HPV tipe high risk yang dapat menyebabkan kanker serviks. Total sampel sebanyak 78 diisolat DNAl yang berasal dari FFPE, apusan serviks dan jaringan segar kanker serviks yang diperoleh dari RSUP. Dr. M Djami., Padang dan RSUD. Achmad Arifin, Pekanbaru. Deteksi DNA HPV dilakukan dengan menggunakan Polymerase Chain Reaction (PCR) menggunakan primer universal GP5+/6+. Tipe HPV yang diidentifikasi dengan metode PCR dengan primer spesifik. Total tipe sampel yang didapat bervariasi dengan konsentrasi antara 0,9-645 ng/ml dengan kemurnian DNA sesuai dengan kemurnian yang ditetapkan untuk amplifikasi PCR. Hasil penelitian dari 78 sampel penderita kanker serviks, 42 sampel (54 %) teridentifikasi DNA HPV. HPV type 18 lebih mendominasi dan disusul HPV type 16 dibandingkan dengan type lainnya yaitu dengan persentase 40,4 % dan 28,5%. HPV type 45 (7,1%), HPV type 52 (2,3%) dan HPV 31 dan HPV type 33 tidak terdeteksi.

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A Multiplexed Human Papillomavirus 16 and 18 Diagnostic Chip for Cervical Cancer in Limited-Resource Settings

Journal of Global Oncology ◽

10.1200/jgo.18.30000 ◽

2018 ◽

Vol 4 (Supplement 1) ◽

pp. 11s-11s ◽

Cited By ~ 1

Author(s):

Winnie S. Wong ◽

Catherine M. Klapperich

Keyword(s):

Cervical Cancer ◽

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

Limit Of Detection ◽

Screening Method ◽

Limited Resource ◽

Clinical Samples ◽

Chain Reaction ◽

Hpv 16 ◽

Polymerase Chain

Abstract 16 Purpose Despite being treatable, cervical cancer is responsible for 300,000 deaths annually and is the second most frequent cause of cancer-related death worldwide. The burden of cervical cancer disproportionately falls on developing countries, where 87% of deaths occur from limited or no resources for screening. Of cervical cancer cases, 99% are caused by human papillomavirus (HPV), with more than 70% of cases from HPV genotypes 16 and 18. We developed a low-cost cervical cancer diagnostic chip to detect HPV 16 and 18 DNA quickly and reliably in limited-resource settings (LRSs). Methods Unlike conventional HPV diagnostics that use polymerase chain reaction and resource-intensive equipment, our chip used isothermal—one temperature—amplification, specifically loop-mediated amplification, and required only a heat source, which made it suitable for LRSs. Amplified DNA was visually detected on a lateral flow strip, which is similar to a pregnancy test, providing a simple yes or no readout. Specific line patterns indicated the presence of HPV 16, 18, both, or neither. Each chip was single use and self-contained to be easily disposed of as biohazard waste, which reduced the risk of contamination and false positives. Results The loop-mediated amplification assay was optimized to detect HPV 16 and 18 simultaneously. Sensitive and specific amplification of cloned HPV 16 and 18 DNA was achieved and confirmed via specific restriction enzyme digests. The lower limit of detection was 104 and 103 copies for HPV 16 and 18, respectively. Clinical samples, namely discarded cervical swab samples, were also tested with the chip. Results were comparable with the gold standard of polymerase chain reaction, which proves that the chip is feasible for clinical samples. Time to result was less than 1 hour, making the chip appropriate for LRSs. Conclusion Our multiplexed HPV 16 and 18 diagnostic chip is clinically relevant and provides a much-needed screening method for LRSs. The chip will increase access to screening for a treatable cancer and provide a faster route to treatment as well as a decrease in deaths from cervical cancer. AUTHORS' DISCLOSURES OF POTENTIAL CONFLICTS OF INTEREST No COIs from the authors.

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Human Papillomavirus (HPV) and HPV 16–Variant Distribution in Vulvar Squamous Cell Carcinoma in Sweden

International Journal of Gynecological Cancer ◽

10.1097/igc.0b013e31826a0471 ◽

2012 ◽

Vol 22 (8) ◽

pp. 1413-1419 ◽

Cited By ~ 20

Author(s):

Gabriella Lillsunde Larsson ◽

Gisela Helenius ◽

Sören Andersson ◽

Fredrik Elgh ◽

Bengt Sorbe ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

Squamous Cell ◽

Survival Rates ◽

Significant Prognostic Factor ◽

Chain Reaction ◽

Hpv 16 ◽

Hpv Types ◽

Polymerase Chain

ObjectiveTo investigate the human papillomavirus (HPV) and HPV type 16–variant distribution in a series of vulvar squamous cell carcinomas (VSCC) and to evaluate the impact of HPV and HPV 16–variant on prognosis.MethodsA series of 133 patients who had a diagnosis of VSCC (1983-2008) was selected for the study. Detection of 11 high-risk HPV types (16, 18, 31, 33, 39, 45, 51, 52, 56, 58, and 59) and 2 low-risk HPV types (6 and 11) was performed with real-time polymerase chain reaction. Samples positive for HPV 16 were further analyzed for variant determination of 7 positions in theE6gene with polymerase chain reaction and pyrosequencing.ResultsForty (30.8%) of 130 tumors were found to be HPV positive. Human papillomavirus type 16 was found in 31 cases, HPV 18 was found in 2 cases, HPV 33 was found in 5 cases, and HPV 56 and HPV 59 were found in one case each. All but one tumor harboring HPV 16 were of European linage, and the 3 most common variants were E-p (n = 13), E-G350 (n = 7), and E-G131 (n = 5). HPV positivity was associated with the basaloid tumor type and occurred in significantly younger patients. Overall and recurrence-free survival rates were better in HPV-positive cases, but after correction for age and tumor size, HPV status was no longer an independent and significant prognostic factor. The survival rates of the various HPV 16 variants were not significantly different, but there was a trend of worse outcome for the E-G131–variant group.ConclusionsHuman papillomavirus positivity of 30.8% is similar to other reports on VSCC. To our knowledge, this first variant determination of HPV 16 in vulvar carcinoma in a Swedish cohort indicated that the variant E-G131 may have an increased oncogenic potential in patients with VSCC.

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The Impact of the Distribution of Human Papillomavirus Types and Associated High-Risk Lesions in a Colposcopy Population for Monitoring Vaccine Efficacy

Archives of Pathology & Laboratory Medicine ◽

10.5858/2008-132-54-tiotdo ◽

2008 ◽

Vol 132 (1) ◽

pp. 54-60

Author(s):

Nick A. Antonishyn ◽

Greg B. Horsman ◽

Rod A. Kelln ◽

Jasdeep Saggar ◽

Alberto Severini

Keyword(s):

Polymerase Chain Reaction ◽

Human Papillomavirus ◽

Cervical Intraepithelial Neoplasia ◽

Real Time ◽

Intraepithelial Neoplasia ◽

Chain Reaction ◽

Hpv 16 ◽

Hpv Types ◽

Polymerase Chain ◽

Hpv 18

Abstract Context.—Impact studies of the new human papillomavirus (HPV) vaccines will be biased unless local baseline distribution studies are conducted. Vaccine cross protection for other important oncogenic HPV types and the emergence of potential genotype replacements require the knowledge of the prevaccine epidemiology of HPV. Objective.—To determine the prevaccine distribution of HPV types in Saskatchewan, using a subpopulation of women referred to a colposcopy clinic. Design.—One thousand three hundred fifty-five specimens obtained during colposcopic examination were typed for HPV using L1 or E1 gene polymerase chain reaction and direct sequencing. HPV-16 and HPV-31 infections were confirmed with real-time E6 polymerase chain reaction. Indeterminate samples were analyzed using Luminex technology. Correlations of the HPV type and histology were examined for statistical significance. Results.—The most commonly identified genotype in patients with cervical intraepithelial neoplasia grade 2 or worse was HPV-16 (46.7%) followed by HPV-31 (14.7%) and then HPV-18 (3.9%). Fifteen of 330 specimens that were positive for HPV-16 or HPV-31 were further resolved to be mixed HPV-16/HPV-31 infections by real-time polymerase chain reaction. The risk of cervical intraepithelial neoplasia associated with HPV-18 infection (0.4–1.7) is substantially lower than with either HPV-16 (3.6–11.0) or HPV-31 (1.8–12.6). Conclusions.—HPV-31 is contributing significantly to the proportion of women with cervical intraepithelial neoplasia in our population and shows a higher prevalence than HPV-18 in high-grade lesions. The clinical significance of HPV-31 may be underestimated and its continued significance will depend on the level of cross protection offered by the new vaccines.

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