Human Papillomavirus Genotyping as a Tool for Cervical Cancer Prevention: From Commercially Available Human Papillomavirus DNA Test to Next-Generation Sequencing

Dias, Tauana Christina; Longatto‐Filho, Adhemar; Campanella, Nathália C.

doi:10.2144/fsoa-2019-0159

Cited by 5 publications

(3 citation statements)

References 53 publications

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“…1. The HPV test is based on the search for 12-14 types of viruses that have a greater oncogenic potential [165]; and 2. The HPV test does not discriminate between transient infections and persistent and productive infections [165].…”

Section: Specimen Adequacymentioning

confidence: 99%

Genetic Screening of Cervical Cancer

Comparetto¹,

Borruto²

2021

obm genet

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Medical genetics plays an important role in the screening and prevention of numerous diseases. Thus, it is important to develop effective screening and prevention programs and improve the assessment of the susceptibility of diseases. The development of screening and prevention programs depends on the identification of early biomarkers (including functional and behavioral) for the risk and onset of the disease, and such programs need to be designed according to internationally accepted criteria. Cervical cancer represents a very relevant disease from the health and social perspective; around 528,000 new cases are diagnosed every year globally, of which, 85% are from developing countries, representing almost 12% of all cancers in females. Substantial reductions in the incidence of and mortality from cervical cancer have been observed after the introduction of prevention campaigns with the implementation of cervical screening programs through Papanicolaou (Pap) tests and, in particular, following the introduction of organized programs which guarantee a high level of screening coverage, as well as, the quality and continuity of diagnostic-therapeutic procedures. It is estimated that Pap smear screening every 3-5 years provides 80% protection against the onset of cancer. Advances in diagnostic techniques, particularly the development of easy-to-use molecular genetic tests, are replacing the use of the established Pap smear as a screening tool. This is possible owing to the discovery in 1975 that some cellular morphological changes (koilocytosis) were related to the presence of a Human Papillomavirus (HPV) infection. The HPV test is performed on a small sample of cells taken from the cervix, similar to the Pap test; however, it is not a morphological exam but a molecular biology exam that detects the presence of HPV by identifying its deoxyribonucleic acid (DNA) or messenger ribonucleic acid (mRNA). The results of numerous experimental studies have demonstrated a greater sensitivity of this test compared to the sensitivity of the traditional Pap test. However, the HPV test has a lower specificity due to two main factors: 1) The HPV test is based on the search for the types of viruses that have a greater oncogenic potential, and 2) It does not discriminate between transient infections and persistent and productive infections. The most widely used molecular tests are based on the search for HPV sequences and genotyping using molecular biology techniques, such as direct hybridization, qualitative polymerase chain reaction (PCR), and viral nucleotide sequencing.

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Section: Specimen Adequacymentioning

confidence: 99%

Genetic Screening of Cervical Cancer

Comparetto¹,

Borruto²

2021

obm genet

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“…Human papillomavirus is the most studied and well-known. Human papillomavirus has around 150 different types (6). Real-time PCR can distinguish nearly related sequences by combining simultaneous PCR amplification with extensive computer analysis of the collected kinetics data (7).…”

Section: Introductionmentioning

confidence: 99%

Detection and Genotyping of Papillomavirus by Real-Time PCR in Iraqi Patients

Al-Hadeithi

Jasim

Salahldina

2022

Arch Clin Infect Dis

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Background: Cervical cancer (CC) is linked to human papillomavirus (HPV). Globally, the prevalence and genotype distribution differ significantly. Objectives: The goal of this study was to find HPV 14, 16, 18, and 45 genotypes in urogenital swabs by using a real-time PCR amplification test for quantitative genotyping of HPV DNA types 16, 18, and 45 and for simultaneous quantitative detection of HPV DNA types 31, 33, 35, 39, 51, 52, 56, 58, 59, 66, and 68, for a total of 14 HPV genotypes. Methods: This case-control study included 86 cervical swabs from Iraqi women referred by the Al-Yarmook teaching hospital in Baghdad, Iraq. The ages of cases varied from 23 to 70 years and specimens were obtained between March 2020 and March 2021. The DNA was extracted for molecular assay. Fourteen HPV genotypes were detected using real-time PCR (16, 18, 45, 31, 33, 35, 39, 51, 52, 56, 58, 59, 66, and 68). The detection protocol was based on the commercial Kit V31-100/F FRT as follows. For each sample reaction, 10х(N+1) μL of PCR-mix-1-FRT HPV 14 was added into a new tube. Then, 5.0х(N+1) μL of PCR-mix-2 buffer and 0.5х(N+1) μL of TaqF DNA polymerase were added. The tubes were vortexed. Finally, the prepared tubes added 10 μL of DNA samples from test or control samples. The statistical analysis was conducted using the statistical package for SPSS and Excel 2016 software. Results: Genotype 16 had the highest frequency, followed by genotypes 45 (22%), 18 (14%), 35 and 59 (6%), 52 and 58 (4%), and 31 (2%), while genotypes 33, 39, 51, 56, 66, and 68 had the lowest frequency (1%). Conclusions: The real-time PCR was efficient for detecting and genotyping HPV-DNA and could help in earlier detection and clinical care of HPV-infected patients by reducing costs and workload.

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“…The HPV test using next generation sequencing (NGS) is a PCR-based analysis that allows for the identification of both HR-and LR-HPV sequencing of the HPV genome and for aligning the results with sequences registered in an HPV database. NGS is a highthroughput sequencing technique that has been widely used to determine the presence of HPV in human specimens using large or small-targeted panels [20][21][22][23][24][25][26][27][28][29][30][31][32][33]. The NGS panel used for HPV detection may be performed with or without a prior PCR.…”

Section: Introductionmentioning

confidence: 99%

Different Methods in HPV Genotyping of Anogenital and Oropharyngeal Lesions: Comparison between VisionArray® Technology, Next Generation Sequencing, and Hybrid Capture Assay

Acquaviva

Visani

Sanza

et al. 2021

JMP

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(1) Background: Human papillomaviruses (HPVs) are known to be related to the development of about 5% of all human cancers. The clinical relevance of HPV infection has been deeply investigated in carcinomas of the oropharyngeal area, uterine cervix, and anogenital area. To date, several different methods have been used for detecting HPV infection. The aim of the present study was to compare three different methods for the diagnosis of the presence of the HPV genome. (2) Methods: A total of 50 samples were analyzed. Twenty-five of them were tested using both next generation sequencing (NGS) and VisionArray® technology, the other 25 were tested using Hybrid Capture (HC) II assay and VisionArray® technology. (3) Results: A substantial agreement was obtained using NGS and VisionArray® (κ = 0.802), as well as between HC II and VisionArray® (κ = 0.606). In both analyses, the concordance increased if only high risk HPVs I(HR-HPVs) were considered as “positive”. (4) Conclusions: Our data highlighted the importance of technical choice in HPV characterization, which should be guided by the clinical aims, costs, starting material, and turnaround time for results.

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Human Papillomavirus Genotyping as a Tool for Cervical Cancer Prevention: From Commercially Available Human Papillomavirus DNA Test to Next-Generation Sequencing

Cited by 5 publications

References 53 publications

Genetic Screening of Cervical Cancer

Genetic Screening of Cervical Cancer

Detection and Genotyping of Papillomavirus by Real-Time PCR in Iraqi Patients

Different Methods in HPV Genotyping of Anogenital and Oropharyngeal Lesions: Comparison between VisionArray® Technology, Next Generation Sequencing, and Hybrid Capture Assay

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