Novel Method for Detection, Typing, and Quantification of Human Papillomaviruses in Clinical Samples

Hart, Keith; Williams, O. Martin; Thelwell, Nicola; Fiander, Alison; Brown, Tom; Borysiewicz, L. K.; Gelder, Colin

doi:10.1128/jcm.39.9.3204-3212.2001

Cited by 44 publications

(37 citation statements)

References 27 publications

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“…All of these strategies have advantages and disadvantages, depending on their application (5,10,26). Several consensus PCR systems have been conveniently used in several large-scale epidemiological studies (9,10,19). However, consensus PCR products do not provide practical information for genotyping (26).…”

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confidence: 99%

Development and Clinical Evaluation of a Highly Sensitive DNA Microarray for Detection and Genotyping of Human Papillomaviruses

Oh¹,

Kim

Woo³

et al. 2004

J Clin Microbiol

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Human papillomavirus (HPV) has been found in cervical cancer, tonsillar cancer, and certain types of head and neck cancers. We report on a DNA microarray-based method for the simultaneous detection and typing of HPVs. The genotype spectrum discriminated by this HPV DNA microarray includes 15 high-risk HPV genotypes and 12 low-risk HPV genotypes. The HPV DNA microarray showed high degrees of specificity and reproducibility. We evaluated the performance of the HPV DNA microarray by application to three HPVpositive cell lines (HeLa, Caski, and SiHa cells) and two HPV-negative cell lines (C33A and A549 cells). The HPV DNA microarray successfully identified the known types of HPV present in the cell lines. The detection limit of the HPV DNA microarray was at least 100-fold higher than that of PCR. To assess the clinical applicability of the HPV DNA microarray, we performed the HPV genotyping assay with 73 nonmalignant and malignant samples from 39 tonsillar cancer patients. Twenty-five of the 39 (64.1%) malignant samples were positive for HPV, whereas 3 of 34 (8.8%) nonmalignant samples were positive for HPV. This result shows a preferential association of HPV with tonsillar carcinomas. The correlations of the presence of HPV with the grade of differentiation and risk factors were not significant. Our data show that the HPV DNA microarray may be useful for the diagnosis and typing of HPV in large-scale epidemiological studies.Epidemiological and molecular studies have demonstrated that high-risk types of human papillomavirus (HPV) not only are etiologically related to the development of most cases of uterine cervical carcinoma (2,8,14,16) but also are associated with certain types of carcinomas in the head and neck (7, 11). Until now, more than 100 different HPV genotypes have been identified on the basis of the DNA sequence of the L1, E6, and upstream regulatory regions (3,17,29). The mucosal HPV genotypes are generally classified into low-risk and high-risk groups on the basis of their association with malignant lesions and phylogenetic relationships (13,15,29). Furthermore, it has been demonstrated in tonsillar carcinoma that HPV types 16 and 33 express the E6 and E7 oncogenes and that transcription is localized in the cancer cells and does not occur in the surrounding stroma (24, 28). Because HPV genotyping information is clinically useful for prognosis and therapy based on the risk type, it is important that the HPV genotype be identified by as sensitive and as specific a method as possible.At present, eight main strategies are used to detect and type various HPVs. All of these strategies have advantages and disadvantages, depending on their application (5, 10, 26). Several consensus PCR systems have been conveniently used in several large-scale epidemiological studies (9, 10, 19). However, consensus PCR products do not provide practical information for genotyping (26). Meanwhile, because it is difficult to design compatible multiple primer sets for genotype-specific PCR, the maximum number of HPVs detectable in...

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confidence: 99%

Development and Clinical Evaluation of a Highly Sensitive DNA Microarray for Detection and Genotyping of Human Papillomaviruses

Oh¹,

Kim

Woo³

et al. 2004

J Clin Microbiol

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“…Recent studies suggest that HPV-16 viral load may be used as a molecular biomarker to improve the prognostic value of HPV testing (8)(9)(10). Other studies found a positive correlation between high HPV-16 load and high-grade squamous intraepithelial lesions (11,12).…”

Section: Introductionmentioning

confidence: 99%

Correlates of Human Papillomavirus Viral Load with Infection Site in Asymptomatic Men

Flores

Nielson

et al. 2008

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Numerous studies have evaluated human papillomavirus (HPV) DNA load in women, especially HPV-16 viral load, and its role in cervical carcinogenicity. Few studies have examined HPV viral load in men, none among asymptomatic men. The aim of the current study is to quantify HPV-16 viral load in male anogenital specimens and to explore its correlates with anatomic sites. Two-hundred and ninety-four specimens from 42 men who tested positive for HPV-16 at one or more anatomic sites were evaluated. HPV DNA was detected with PGMY 09/11 primer and genotyped with reverse line blot assay followed by HPV-16 viral quantification using type-specific real-time PCR assay (TaqMan). The quantitative PCR assay showed a higher sensitivity in HPV-16 viral DNA detection compared with the reverse line blot assay. Viral load varied significantly by anatomic site (P = 0.019). Penile shaft specimens had significantly higher viral load than any other anatomic site evaluated except for the anal canal. HPV-16 viral load was positively correlated between proximal anatomic sites: perianal and anal canal (P = 0.003), perianal and scrotum (P = 0.011), scrotum and glans/corona (P = 0.045), and scrotum and penile shaft (P = 0.037).In conclusion, the penile shaft seemed to be the preferred site for HPV-16 viral replication. Viral load correlation between proximal sites suggested a possible autoinoculation in male HPV transmission. (Cancer Epidemiol Biomarkers Prev 2008;17(12):3573-6)

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“…Many PCR-ELISAs have been developed for the detection, identification, and quantification of a large variety of viruses (2,8,14,23,35,46,47,50,57,62,63,71,75,77,83). In the present study, we have developed and evaluated a PCR-ELISA protocol for the detection and identification of clinically relevant human group A rotavirus G (G1 to G6, G8 to G10, and G12) and P (P [4], P [6], P [8], P [9], and P [14]) genotypes.…”

Section: Discussionmentioning

confidence: 99%

Development of a Microtiter Plate Hybridization-Based PCR-Enzyme-Linked Immunosorbent Assay for Identification of Clinically Relevant Human Group A Rotavirus G and P Genotypes

et al. 2008

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A microtiter plate hybridization-based PCR-enzyme-linked immunosorbent assay (PCR-ELISA) has been used for the detection and identification of a variety of microorganisms. Here, we report the development of a PCR-ELISA for the identification of clinically relevant human rotavirus VP7 (G1 to G6, G8 to G10, and G12) and VP4 (P[4], P[6], P[8], P[9], and P[14]) genotypes. The G and P types of reference human and animal rotavirus strains for which specific probes were available were correctly identified by the PCR-ELISA. In addition, reference strains bearing G or P genotypes for which specific probes were unavailable, such as G11, G14, P[3], P[10], and P[11], did not display any cross-reactivity to the probes. The usefulness of the assay was further evaluated by analyzing a total of 396 rotavirus-positive stool samples collected in four countries: Brazil, Ghana, Japan, and the United States. The results of this study showed that the PCR-ELISA was sensitive and easy to perform without the use of any expensive and sophisticated equipment, the reagents used are easy to obtain commercially and advantageous over multiplex PCR since more than one type-specific probe is used and the selection of probes is more flexible.

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Novel Method for Detection, Typing, and Quantification of Human Papillomaviruses in Clinical Samples

Cited by 44 publications

References 27 publications

Development and Clinical Evaluation of a Highly Sensitive DNA Microarray for Detection and Genotyping of Human Papillomaviruses

Development and Clinical Evaluation of a Highly Sensitive DNA Microarray for Detection and Genotyping of Human Papillomaviruses

Correlates of Human Papillomavirus Viral Load with Infection Site in Asymptomatic Men

Development of a Microtiter Plate Hybridization-Based PCR-Enzyme-Linked Immunosorbent Assay for Identification of Clinically Relevant Human Group A Rotavirus G and P Genotypes

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