scholarly journals Comparison of the Hybrid Capture II Method with a PCR-Based Screening Method Using a Carboxyfluorescein-Labeled Primer for Detecting Human Papillomavirus in Cervicovaginal Liquid-Based Cytology

2020 ◽  
Vol 1 (1) ◽  
pp. 9-18
Author(s):  
Yusuke Saiki ◽  
Yuka Gion ◽  
Asami Nishikori ◽  
Yoshiaki Norimatsu ◽  
Yasuharu Sato
Keyword(s):  
Screening Method ◽  
Human Papillomaviruses ◽  
Screening Methods ◽  
Consensus Primer ◽  
Perfect Agreement ◽  
Diagnostic Reliability ◽  
Pap Tests ◽  
Hybrid Capture ◽  
Hpv Dna ◽  
Liquid Based Cytology

Objective: Human papillomaviruses (HPVs) are DNA viruses, of which over 120 types have been identified. The main screening methods for HPV-DNA include the hybrid capture II (HC-II) and polymerase chain reaction (PCR) assays. Liquid-based cytology (LBC) is a high-quality technique developed to improve the diagnostic reliability of traditional Papanicolaou tests (Pap tests). However, relatively few studies have compared the efficacy of PCR and HC-II assays using cervicovaginal LBC specimens. In this study, we conducted a comparative analysis with results derived from the HC-II assay to assess whether a PCR-based assay using a novel carboxyfluorescein (FAM)-labeled primer could be applied to cervicovaginal LBC specimens. Methods and Results: We analyzed 59 specimens diagnosed as atypical squamous cells of undetermined significance (ASCUS) by Pap tests. After extracting DNA from cervicovaginal LBC specimens, we performed PCR using a FAM-labeled consensus primer, and then conducted fragment analysis to confirm the results. The value of the kappa statistic measuring the agreement between the PCR and HC-II results was 0.8557, or “almost perfect agreement.” Conclusion: Our novel HPV-PCR assay can be successfully applied to cervicovaginal LBC specimens for the detection of HPV subtypes.

scholarly journals Detection of human papillomavirus in cervical cell specimens by hybrid capture and PCR with different primers.

2006 ◽  
Vol 53 (3) ◽  
pp. 603-607 ◽  
Author(s):  
Slawa Szostek ◽  
Malgorzata Klimek ◽  
Barbara Zawilinska ◽  
Janusz Rys ◽  
Jolanta Kope ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
High Risk ◽  
Consensus Primer ◽  
Hybrid Capture ◽  
Hpv Dna ◽  
Cervical Smears ◽  
Capture Assay ◽  
Cervical Cell ◽  
Target Dna ◽  
Primer Sets

The purpose of this study was to compare hybrid capture assay with PCRs using different primers for the L1, E6-E7 regions for the detection of human papillomavirus (HPV) genome. One hundred twenty-five cervical smears with normal (n=42) and abnormal (n=83) cytology were investigated. Those at high-risk for HPV were studied by hybridization antibody capture assay and PCR with the pU-1M/pU-2R primers. Target DNA from the HPV L1 region was amplified by SPF10 primer set and home-PCR with MY09/MY11 primers. The presence of HPV DNA in cervical smears was detected by SPF10 (in 72% of cases), MY09/MY11 (58%), hybrid capture (55%) and pU-1M/pU-2R (39%). Results obtained with the SPF10 and MY09/MY11 consensus primer sets as well as hybrid capture and pU-1M/pU-2R specific for high-risk types differed significantly (chi2, P

scholarly journals Evaluation of a liquid bead array system for high-risk human papillomavirus detection and genotyping in comparison with Hybrid Capture II, DNA chip and sequencing methods

2011 ◽  
Vol 60 (2) ◽  
pp. 162-171 ◽  
Author(s):  
Eun Jung Cho ◽  
Jin Hwan Do ◽  
Yoon Sun Kim ◽  
Sumi Bae ◽  
Woong Shick Ahn
Keyword(s):  
Human Papillomavirus ◽  
High Risk ◽  
Intraepithelial Neoplasia ◽  
Dna Chip ◽  
Detection Methods ◽  
Specific Information ◽  
Perfect Agreement ◽  
Hybrid Capture ◽  
Hpv Dna ◽  
Genotype Information

Since persistent infection with high-risk human papillomavirus (HPV) is a known cause of high-grade cervical intraepithelial neoplasia and cervical cancer, several HPV DNA detection methods have been developed during the last decade. The Hybrid Capture II (HCII) assay, which allows detection of 13 high-risk HPVs, has been well validated; however, it does not provide any genotype-specific information. The oncogenic activity of HPV is dependent on its genotype. The prophylactic effects of HPV vaccines are based on L1 virus-like particles and are limited mainly to infections corresponding to the HPV type used to develop the immunogen. Therefore, accurate detection and genotyping are important for treatment as well as screening. A newly developed HPV genotyping system using a liquid bead array was evaluated with 286 cervical samples and the results were compared to two commercially available methods, i.e. the HCII and HPV DNA chip assays, and sequencing. The sensitivity for detection of high-risk HPV was 85.3 % (HCII), 94.7 % (DNA chip) and 99.0 % (liquid bead array). The liquid bead array showed almost perfect agreement (κ=0.95) with genotype information confirmed by sequencing, while substantial agreement (κ=0.8) was observed between DNA chip and sequencing. Furthermore, the liquid bead array had superior detection of 26 HPVs (16 high-risk and 10 low-risk types) and has proven to be as accurate as sequencing in identifying individual HPV types, even in cases with multiple HPV infections.

scholarly journals A Comparative Study of Manual Liquid-Based Cytology with the Conventional Pap Smear for Cervical Cancer Screening in a Tertiary Care Hospital of Mandya, Karnataka

2021 ◽  
Vol 8 (16) ◽  
pp. 1009-1014
Author(s):  
Manjunath M.R ◽  
Sheetal Sheetal
Keyword(s):  
Cervical Cancer ◽  
Pap Smear ◽  
Invasive Cervical Cancer ◽  
Tertiary Care ◽  
Human Papillomaviruses ◽  
Care Hospital ◽  
Cancer Cervix ◽  
Hpv Dna ◽  
Bethesda System ◽  
Liquid Based Cytology

BACKGROUND A long pathological process for investigation of precursor lesion squamous intraepithelial lesion (SIL) leads to invasive cervical cancer. This SIL can be detected much earlier before the lesion progresses to invasive cancer. For greater than fifty years, screening for cancer cervix was done by conventional scrape smears and stained by Papanicolaou [Pap] stain but conventional Pap smears (CPAP) have been reported to have low sensitivity. To overcome these drawbacks manual liquid-based cytology [MLBC] was introduced. The objective of this study was to screen females for cervical cancer using CPAP and MLBC techniques and compare the results of these techniques. METHODS Cervical cytology samples were obtained from 120 non-pregnant females through specialised Uprep cytobrush with a detachable head. Thus, obtained samples were first smeared onto a clean glass slide for CPAP smear, and the whole head to cytobrush was dropped into the specialised Uprep liquid preservative medium and processed by using Uprep Cytospin machine to obtain MLBC smears. Both the smears were stained by conventional Pap stain and reported according to 2014 Bethesda system. RESULTS In this study, the CPAP method had a greater number of unsatisfactory smears than that of MLBC method which was statistically significant. MLBC identified more number of intraepithelial lesions when compared to CPAP and MLBC had an increased detection rate [IDR] of 73.68 % over CPAP. CONCLUSIONS Analysis of our results showed that MLBC had more advantages over CPAP. Since the cost effective MLBC has an improved rate of detection of abnormal lesions, MLBC can be used as a routine technique for screening of cancer cervix in India. Also, MLBC offers an important advantage of performing both human papillomaviruses deoxyribonucleic acid (HPV DNA) test and cytological analysis on a single sample. KEYWORDS Cervical Cancer, Conventional Pap Smear, Manual Liquid Based Cytology, Human Papilloma Virus (HPV DNA), Bethesda System

scholarly journals The Accuration of LiquidBased Cytology and HPV DNA Test Combination as Precervical Cancer Lesion Screening

2017 ◽  
pp. 241
Author(s):  
Junita Indarti ◽  
Yuven S Pratama
Keyword(s):  
False Negative ◽  
Screening Method ◽  
Cross Sectional Study ◽  
Health Clinic ◽  
Cross Sectional ◽  
Dna Test ◽  
Hpv Dna ◽  
Liquid Based Cytology ◽  
Cancer Lesion ◽  
Hpv Dna Test

Objective: To investigate the accuracy of liquid-based cytology, HPV DNA test, and the combination of liquid-based cytologyband HPV DNA test, compared to histopathology as the gold standard of precervical cancer lesion screening. Methods: This was a cross-sectional study. The medical records of patients who came to the Women’s Health Clinic of Dr. Cipto Mangunkusumo Hospital during the period of July 2013 to December 2015 were evaluated. Results: The high risk type HPV DNA is detected in 76% CIN 1, 88.46% CIN 2, and 84.21 CIN 3 in histopathology results. The accuracy of liquid-based cytology; sensitivity 88.54%, specificity35.71%, PPV 75.89%, and NPV 57.69%. The accuracy of HPV DNA; sensitivity 81.25%, specificity 78.57%, PPV 89.66%, and NPV 64.71%. The accuracy of combination: sensitivity 94.79%, specificity 35.71%, PPV 77.12%, and NPV 75%. Conclusion: The addition of HPV DNA test increased the sensitivity from 88.54% to 94.79% because of decreasing of false negative of liquid-based cytology. This thing has showed that the combination of liquid-based cytology and HPV DNA test could the one of the option of precervical cancer lesion screening method, especially in secondary or tertier health center in Indonesia. Keywords: accuracy test, HPV DNA, liquid-based cytology, precervical cancer lesion, precervical cancer lesion screening

Human Papillomavirus Genome based Detection and Typing: A Holistic Molecular Approach

2019 ◽  
Vol 19 (4) ◽  
pp. 237-246 ◽  
Author(s):  
Abhilasha Gautam ◽  
Mallikarjuna R. Gedda ◽  
Madhukar Rai ◽  
Shyam Sundar ◽  
Jaya Chakravarty
Keyword(s):  
Human Papillomavirus ◽  
High Risk ◽  
Genome Structure ◽  
Cost Effective ◽  
Detection Sensitivity ◽  
Screening Methods ◽  
Consensus Primer ◽  
Hpv Dna ◽  
Primer Sets ◽  
Consensus Primers

Human Papillomavirus (HPV) is a species specific double-stranded DNA virus infecting human cutaneous or mucosal tissues. The genome structure of HPV is extremely polymorphic hence making it difficult to discriminate between them. HPV exhibits numerous dissimilar types that can be subdivided into high-risk (HR), probably high-risk and low-risk (LR), causing numerous types of cancers and warts around the genital organs in humans. Several screening methods are performed in order to detect cytological abnormalities and presence or absence of HPV genome. Currently available commercial kits and methods are designed to detect only a few HR/LR-HPV types, which are expensive adding to the economic burden of the affected individual and are not freely available. These gaps could be minimized through Polymerase Chain reaction (PCR) method, which is a gold standard and a cost-effective technique for the detection of most HPV (both known and unknown) types by using specific consensus primers in minimal lab setup. In this context, numerous studies have validated the effectiveness of different sets of consensus primers in the screening of HPVs. Numerous consensus primers, such as E6, E6/E7, GP-E6/E7, MY09/11, GP5+/GP6+, SPF10, and PGMY09/11 have been developed to detect the presence of HPV DNA. In addition, HPV detection sensitivity could be achieved through consensus primer sets targeting specific ORF regions like L1 and E6, which may finally assist in better diagnosis of several unknown HR-HPVs. The present review, provides a summary of the available methods, kits and consensus primer sets for HPV genome based detection, their advantages and limitations along with future goals to be set for HPV detection.

scholarly journals Clinical Prediction Based on HPV DNA Testing by Hybrid Capture 2 (HC2) in Combination with Liquid-based Cytology (LBC)

2013 ◽  
Vol 14 (2) ◽  
pp. 903-907 ◽  
Author(s):  
Pairoj Junyangdikul ◽  
Watcharaporn Tanchotsrinon ◽  
Jira Chansaenroj ◽  
Pornjarim Nilyaimit ◽  
Chidchanok Lursinsap ◽  
...  
Keyword(s):  
Clinical Prediction ◽  
Dna Testing ◽  
Hybrid Capture ◽  
Hpv Dna ◽  
Hpv Dna Testing ◽  
Hybrid Capture 2 ◽  
Liquid Based Cytology

Prevalence and typing of HPV DNA by hybrid capture II in women with ASCUS, ASC-H, LSIL, and AGC on ThinPrep® Pap tests

2004 ◽  
Vol 30 (6) ◽  
pp. 426-432 ◽  
Author(s):  
Leslie R. Rowe ◽  
William Aldeen ◽  
Joel S. Bentz
Keyword(s):  
Pap Tests ◽  
Hybrid Capture ◽  
Hpv Dna

Comparative Evaluation of the HPV28 Detection and HPV DNA Chip Test for Detecting and Genotyping Human Papillomaviruses

2013 ◽  
Vol 3 (4) ◽  
pp. 234 ◽  
Author(s):  
Eunsim Shin ◽  
Heojin Bae ◽  
Wan-Keun Song ◽  
Sun-Kyung Jung ◽  
Yoo-Sung Hwang
Keyword(s):  
Comparative Evaluation ◽  
Human Papillomaviruses ◽  
Dna Chip ◽  
Hpv Dna

scholarly journals Development and validation of a multiplex qPCR assay for detection and relative quantification of HPV16 and HPV18 E6 and E7 oncogenes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexia Bordigoni ◽  
Anne Motte ◽  
Hervé Tissot-Dupont ◽  
Philippe Colson ◽  
Christelle Desnues
Keyword(s):  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Papillomaviruses ◽  
Molecular Techniques ◽  
Clinical Samples ◽  
Gene Encoding ◽  
Hpv Dna ◽  
E6 And E7 ◽  
High Risk Hpv ◽  
Development And Validation

AbstractHuman papillomaviruses (HPV) play a key role in promoting human anogenital cancers. Current high-risk HPV screening or diagnosis tests involve cytological or molecular techniques mostly based on qualitative HPV DNA detection. Here, we describe the development of a rapid quantitative polymerase chain reaction (qPCR) detection test of HPV16 and HPV18 oncogenes (E6 and E7) normalized on human gene encoding GAPDH. Optimized qPCR parameters were defined, and analytical specificities were validated. The limit of detection was 101 for all genes tested. Assay performances were evaluated on clinical samples (n = 96). Concordance between the Xpert HPV assay and the triplex assay developed here was 93.44% for HPV16 and 73.58% for HPV18. HPV co-infections were detected in 15 samples. The systems developed in the present study can be used in complement to traditional HPV tests for specifically validating the presence of HPV16 and/or HPV18. It can also be used for the follow-up of patients with confirmed infection and at risk of developing lesions, through the quantification of E6 and E7 oncogene expression (mRNA) normalized on the GAPDH expression levels.

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

2021 ◽  
Vol 2 (1) ◽  
pp. 29-41
Author(s):  
Giorgia Acquaviva ◽  
Michela Visani ◽  
Viviana Sanza ◽  
Antonio De Leo ◽  
Thais Maloberti ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Turnaround Time ◽  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Next Generation ◽  
Hybrid Capture ◽  
Capture Assay ◽  
Anogenital Area ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

(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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