Development of a Sensitive and Specific Assay Combining Multiplex PCR and DNA Microarray Primer Extension To Detect High-Risk Mucosal Human Papillomavirus Types
Tarik Gheit,Stefano Landi,Federica Gemignani,Peter J.F. Snijders,Salvatore Vaccarella,Silvia Franceschi,Federico Canzian,Massimo Tommasino +7 more
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TLDR
A novel one-shot method for the detection and typing of 19 mucosal high-risk (HR) HPV types that combines two different techniques: multiplex PCR with HPV type-specific primers for amplification of viral DNA and array primer extension (APEX) for typing.Abstract:
The importance of assays for the detection and typing of human papillomaviruses (HPVs) in clinical and epidemiological studies has been well demonstrated. Several accurate methods for HPV detection and typing have been developed. However, comparative studies showed that several assays have different sensitivities for the detection of specific HPV types, particularly in the case of multiple infections. Here, we describe a novel one-shot method for the detection and typing of 19 mucosal high-risk (HR) HPV types (types 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, and 82). This assay combines two different techniques: multiplex PCR with HPV type-specific primers for amplification of viral DNA and array primer extension (APEX) for typing. This novel method has been validated with artificial mixtures of HPV DNAs and clinical samples that were already analyzed for the presence of mucosal HPV types by a different consensus PCR method, i.e., GP5+/GP6+. Our data showed a very good agreement between the results from the multiplex PCR/APEX assay and those from the GP5+/GP6+ PCR (overall rates of HPV positivity, 63.0 and 60.9%, respectively). Whereas the GP5+/GP6+ PCR was slightly more sensitive for the detection of HPV type 16 (HPV-16), multiplex PCR-APEX found a higher number of infections with HPV-33, HPV-53, and multiple HPV types. These favorable features and the high-throughput potential make our present novel assay ideal for large-scale clinical and epidemiological studies aimed at determining the spectrum of mucosal HR HPV types in cervical specimens.read more
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Basic Concepts of Microarrays and Potential Applications in Clinical Microbiology
Melissa B. Miller,Yi-Wei Tang +1 more
TL;DR: This review highlights uses of microarray technology that impact diagnostic microbiology, including the detection and identification of pathogens, determination of antimicrobial resistance, epidemiological strain typing, and analysis of microbial infections using host genomic expression and polymorphism profiles.
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TLR9 expression and function is abolished by the cervical cancer-associated human papillomavirus type 16.
Uzma Hasan,Elizabeth E.M. Bates,Fumihiko Takeshita,Alexandra Biliato,Rosita Accardi,Véronique Bouvard,Mariam Mansour,Isabelle Vincent,Lutz Gissmann,Thomas Iftner,Mario Sideri,Frank Stubenrauch,Massimo Tommasino +12 more
TL;DR: A novel mechanism used by HPV16 to suppress the host immune response by deregulating the TLR9 transcript is revealed, providing evidence that abolishing innate responses may be a crucial step involved in the carcinogenic events mediated by HPVs.
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Gene expression analysis of preinvasive and invasive cervical squamous cell carcinomas identifies HOXC10 as a key mediator of invasion.
Yali Zhai,Rork Kuick,Bin Nan,Ichiro Ota,Stephen J. Weiss,Cornelia L. Trimble,Eric R. Fearon,Kathleen R. Cho +7 more
TL;DR: Findings support a key role for the HOXC10 homeobox protein in cervical cancer progression and suggest other genes with differential expression in invasive SCC versus HSIL may contribute to tumor progression or may be useful as markers for cancer diagnosis or progression risk.
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Immunogenicity and HPV infection after one, two, and three doses of quadrivalent HPV vaccine in girls in India: a multicentre prospective cohort study
Rengaswamy Sankaranarayanan,Priya R. Prabhu,Michael Pawlita,Tarik Gheit,Neerja Bhatla,Richard Muwonge,Bhagwan M. Nene,Pulikottil Okuru Esmy,Smita Joshi,Usha Rani Reddy Poli,Parimal Jivarajani,Yogesh Verma,Eric Zomawia,Maqsood Siddiqi,Surendra S Shastri,Kasturi Jayant,Sylla G. Malvi,Eric Lucas,Angelika Michel,Julia Butt,Janki Mohan Babu Vijayamma,Subha Sankaran,Thiraviam Pillai Rameshwari Ammal Kannan,Rintu Varghese,Uma Divate,Shila Thomas,Geeta Joshi,Martina Willhauck-Fleckenstein,Tim Waterboer,Martin Müller,Peter Sehr,Sanjay Hingmire,Alka Kriplani,Gauravi A Mishra,Sharmila Pimple,Radhika Jadhav,Catherine Sauvaget,Massimo Tommasino,Madhavan Radhakrishna Pillai +38 more
TL;DR: Despite the limitations imposed by the suspension of the HPV vaccination, the findings lend support to the WHO recommendation of two doses, at least 6 months apart, for routine vaccination of young girls.
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Abundance of Multiple High-Risk Human Papillomavirus (HPV) Infections Found in Cervical Cells Analyzed by Use of an Ultrasensitive HPV Genotyping Assay
Markus Schmitt,Bolormaa Dondog,Bolormaa Dondog,Tim Waterboer,Michael Pawlita,Massimo Tommasino,Tarik Gheit +6 more
TL;DR: The TS-PCR-MPG assay significantly increased the rate of detection of HPV DNA and the number of infections with multiple HPV types detected and demonstrated that the prevalence of low-copy-number HPV infections in the anogenital tract may be strongly underestimated by conventional HPV amplification methods, especially in cases of multiple infections.
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Practical statistics for medical research
TL;DR: Practical Statistics for Medical Research is a problem-based text for medical researchers, medical students, and others in the medical arena who need to use statistics but have no specialized mathematics background.
Journal ArticleDOI
Epidemiologic Classification of Human Papillomavirus Types Associated with Cervical Cancer
Nubia Muñoz,F. Xavier Bosch,Silvia de Sanjosé,Rolando Herrero,Xavier Castellsagué,Keerti V. Shah,Peter J.F. Snijders,Chris J.L.M. Meijer +7 more
TL;DR: In addition to HPV types 16 and 18, types 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82Should be considered carcinogenic, or high-risk, types, and types 26, 53, and 66 should be considered probably carcinogenic.