Detection and Typing of Genital Human Papillomaviruses

Ting, Yi Tian; Manos, M. Michele

doi:10.1016/b978-0-12-372180-8.50046-9

Cited by 223 publications

(180 citation statements)

References 6 publications

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“…450 bp fragment of the L1 region was used for PCR. 7,8 HPV type was identified by restriction fragment length polymorphisms (RFLPs) using endonucleases RsaI and DdeI as previously described. 8 Ambiguous RFLPs were resolved by direct sequencing of PCR amplicons.…”

Section: Methodsmentioning

confidence: 99%

Biases in human papillomavirus genotype prevalence assessment associatedwith commonly used consensus primers

Chan

Cheung

Tam

et al. 2005

Intl Journal of Cancer

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Consensus primers targeting human papillomaviruses (HPVs) have biases in sensitivity toward certain HPV types. We applied 3 primer sets (GP51/61, MY09/11, PGMY09/11) in parallel on 120 Chinese cervical cancer specimens. GP51/61 exhibited a poor sensitivity for HPV52, for which the prevalence among squamous cell cervical cancer was underestimated from 14.6% to 0%. The fact that HPV52 should rank second in prevalence among squamous cell cervical carcinoma in Hong Kong could be missed if GP51/61, a worldwide commonly used primer set, was selected for HPV detection. Biases in HPV type-specific sensitivity may result in misprioritization of vaccine candidates. ' 2005 Wiley-Liss, Inc.Key words: human papillomavirus; detection; genotype; vaccine; Chinese; Hong Kong; PCR Cervical cancer is the second most common cancer in women worldwide. Strong and consistent evidence accumulated over the past 2 decades confirms the aetiologic role of human papillomaviruses (HPVs) and provides a strong impetus for developing HPV vaccines to prevent cervical cancer. More than 100 HPV types have been identified, and at least 30 have been found in cervical cancers. 1 Given the diversity of HPV types and the largely typespecific immunity after natural infections, 2-4 it is important to delineate the prevalence of different HPV types found in cervical cancers so as to guide the selection of vaccine candidates. Most studies on HPV have employed consensus primers with an intention to cover a broad spectrum of HPV types. With the more than 10% sequence variation between HPV types, biases in sensitivity of a given primer set toward certain types could happen. In our study, we examined the influence of detection methods on assessing the prevalence of HPVs and thus their priority as cervical cancer vaccine candidates. Material and methodsA total of 120 cervical cancer specimens (105 fresh frozen and 15 paraffin embedded; 89 squamous cell carcinoma, 26 adenocarcinoma, 4 adenosquamous carcinoma and 1 lymphoepitheliod carcinoma) collected from Hong Kong Chinese aged 26-84 years (mean 55 years; SD 13.9) were examined. Total DNA was extracted by the QIAamp DNA mini kit (QIAGEN, Hilden, Germany) and with the quality of extracted preparations confirmed by beta-globin PCR. 5 The clinical materials were collected with a written informed consent. Our study was approved by the local institutional ethics committee, and the human experimentation guidelines of the local institute were followed in the conduct of our study.HPV detection and typing was accomplished by 3 different methods in parallel. In the first method, HPV DNA was amplified by the GP51/61 primers that target an approx. 150 bp fragment of the L1 region. 5,6 The HPV type was identified by direct sequencing of PCR amplicons. In the second method, the MY09/ 11 primers that target an approx. 450 bp fragment of the L1 region was used for PCR. 7,8 HPV type was identified by restriction fragment length polymorphisms (RFLPs) using endonucleases RsaI and DdeI as previously described. 8 Ambiguous RFLPs w...

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Section: Methodsmentioning

confidence: 99%

Biases in human papillomavirus genotype prevalence assessment associatedwith commonly used consensus primers

Chan

Cheung

Tam

et al. 2005

Intl Journal of Cancer

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“…The DNA fragments for HPV types 16 (98 bp) and 18 (80 bp) were from the E6-E7 exon region and were about the same size (17). The culture medium containing the HPV fragments was prepared by diluting 1:20 the PCR-amplified HPV products of previous PCR runs with culture medium for a final concentration of about 0.01 g DNA/L.…”

Section: Treatment Of Mouse Embryosmentioning

confidence: 99%

“…The culture medium containing the HPV fragments was prepared by diluting 1:20 the PCR-amplified HPV products of previous PCR runs with culture medium for a final concentration of about 0.01 g DNA/L. The previous PCR runs were targeted at the HPV gene sequences for types 16 and 18 using paraffin embedded cervical condyloma tissues as DNA templates and published primers (17)(18)(19). The PCR-amplified products were confirmed to contain HPV DNA fragments for the two HPV types using 5% polyacrylamide gel electrophoresis and ethidium bromide staining techniques prior to their usage.…”

Section: Treatment Of Mouse Embryosmentioning

confidence: 99%

Human papilloma virus DNA exposure and embryo survival is stage-specific

Henneberg

Patton

Jacobson

et al. 2006

J Assist Reprod Genet

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Purpose: Human papillomavirus (HPV) has been shown to disrupt late-stage implanting embryos. The objectives were (a) to assess the development of early embryos exposed to HPV DNA and (b) to analyze the blastocyst hatching process after HPV exposure. Methods:The study involved exposing two-cell and 4-8-cell mouse embryos to DNA fragments from either HPV type 16, type 18 or DQA1 (control). The embryos were incubated for 120 h and assessed. Results: HPV 16 and 18 inhibited two-cell embryo development. In contrast, delaying the exposure of HPV DNA until the 4-8-cell stage resulted in further embryonic development. There was 25.9% less blastocyst formed with HPV 16 exposure. Additionally, there were 25.9-31.8% more degenerated embryos with HPV 16 exposure. Conclusions:The study demonstrated embryo stage-specific effects of HPV on early development. The results suggested HPV exposure was linked to two-cell embryo demise and delaying the exposure of HPV until later embryo stages permitted embryo development. HPV 16 was shown to decrease blastocyst formation while HPV 18 inhibited the blastocyst hatching process.

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“…highly conserved L1 region (encodes the major capsid protein) among a broad spectrum of HPV types. 12 Positive samples by the GP set were also tested with type-specific primer sets. Human papillomavirus 16-and 18-specific primers were used to amplify 152 and 216 b.p.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

Human papillomavirus associated with bladder carcinoma? Analysis by polymerase chain reaction

et al. 1999

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Background: The aim of the present study was to assess the possible etiologic role of human papillomaviruses (HPV) in bladder tumors. Methods: Forty-two fresh biopsy specimens from different grades and stages of bladder tumor cases and 10 normal bladder mucosa biopsies were studied. Specimens were analyzed by polymerase chain reaction (PCR) with HPV-specific general primer set for the detection of viral DNA. Polymerase chain reaction-positive samples were also tested with HPV 16-and 18-specific primers by the same method. Results: We found two samples (4.8%) containing HPV DNA among the TaG1 bladder tumors. All other specimens, including the control group, were found to be negative by PCR. Neither of the two HPV-positive patients had immune deficiency and/or genital warts. Human papillomavirus 16 was detected by type-specific primers in one sample, but the other HPV-positive sample could not be typed. Conclusions:The low prevalence of HPV in this and many previous studies does not support an etiologic role of HPV in bladder carcinogenesis. We detected the virus in two early stage tumors, but none was detected in the high-grade samples. However, to clarify the positivity of HPV in these occasional cases, future studies must be designed by using in situ PCR techniques, including samples from tumors and normal bladder mucosa from the same patient.

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Detection and Typing of Genital Human Papillomaviruses

Cited by 223 publications

References 6 publications

Biases in human papillomavirus genotype prevalence assessment associatedwith commonly used consensus primers

Biases in human papillomavirus genotype prevalence assessment associatedwith commonly used consensus primers

Human papilloma virus DNA exposure and embryo survival is stage-specific

Human papillomavirus associated with bladder carcinoma? Analysis by polymerase chain reaction

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