Human Papillomaviruses: Methods and Protocols / Edition 1

by Clare Davy, John Doorbar

Overview

Leading basic researchers and clinical scientists describe in detail a wide variety of established and cutting-edge techniques they have developed to study the lifecycle and biological properties of the human papillomavirus. The authors use these readily reproducible methods, ranging from PCR to propagation of HPV in vitro, to detect and type papillomavirus infections, study the papillomavirus lifecycle, and to produce and functionally analyze papillomavirus proteins. The prools follow the successful Methods in Molecular Medicine™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Product Details

ISBN-13:	9781617375262
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/10/2010
Series:	Methods in Molecular Medicine , #119
Edition description:	Softcover reprint of hardcover 1st ed. 2006
Pages:	496
Product dimensions:	5.98(w) x 9.02(h) x 0.05(d)

Table of Contents

Identification of New Papillomavirus Types.- Identification of HPV Variants.- Hishemical Analysis of Cutaneous HPV-Associated Lesions.- Histological Analysis of Cervical Intraepithelial Neoplasia.- Detection of Papillomavirus Proteins and DNA in Paraffin-Embedded Tissue Sections.- Detection and Quantitation of HPV Gene Expression Using Real-Time PCR.- Analysis of p16INK4a and Integrated HPV Genomes as Progression Markers.- Use of Biomarkers in the Evaluation of CIN Grade and Progression of Early CIN.- HPV DNA Detection and Typing in Cervical Scrapes.- HPV DNA Detection and Typing in Inapparent Cutaneous Infections and Premalignant Lesions.- Establishing HPV-Containing Keratinocyte Cell Lines From Tissue Biopsies.- Using an Immortalized Cell Line to Study the HPV Life Cycle in Organotypic “Raft” Cultures.- Differentiation of HPV-Containing Cells Using Organotypic “Raft” Culture or Methylcellulose.- Propagation of Infectious, High-Risk HPV in Organotypic “Raft” Culture.- Retrovirus-Mediated Gene Transfer to Analyze HPV Gene Regulation and Protein Functions in Organotypic “Raft” Cultures.- The HPV Xenograft Severe Combined Immunodeficiency Mouse Model.- The Cottontail Rabbit Papillomavirus Model of High-Risk HPV-Induced Disease.- Studying the HPV Life Cycle in 3A Trophoblasts and Resulting Pathophysiology.- Replication and Encapsidation of Papillomaviruses in Saccharomyces cerevisiae.- Analysis of the Regulation of Viral Transcription.- Analysis of HPV Transcription by RPA.- Analysis of Regulatory Motifs Within HPV Transcripts.- Detection of HPV Transcripts by Nested RT-PCR.- Analysis of HPV DNA Replication Using Transient Transfection and Cell-Free Assays.- Detection and Quantitation of HPV DNA Replication by Southern Blotting and Real-time PCR.-Analysis of E7/Rb Associations.- Transformation Assays for HPV Oncoproteins.- Analysis of Adeno-Associated Virus and HPV Interaction.- In Vitro Assays of Substrate Degradation Induced by High-Risk HPV E6 Oncoproteins.- Measuring the Induction or Inhibition of Apoptosis by HPV Proteins.- Codon Optimization of Papillomavirus Genes.- Generation of HPV Pseudovirions Using Transfection and Their Use in Neutralization Assays.- Generation and Applications of HPV Pseudovirions Using Vaccinia Virus.

Editorial Reviews

3 Stars from Doody

Reviewer: Rebecca T. Horvat, PhD, D (ABMM)(University of Kansas Medical Center)
Description: This book includes current protocols used by investigators to study human papillomaviruses. The book is one of several in a series that specifically features methods in molecular medicine.
Purpose: According to the authors, this book is designed to be a useful resource for scientists working in the field of papillomavirus research. The book covers a number of different techniques in great detail. In some cases similar techniques are presented from different authors, helping to give a perspective on how these techniques are used in different laboratories. This book would be very useful for those investigators interested in work with human papillomaviruses or oncogenesis.
Audience: This book is targeted at laboratory scientists. It is authored by experts currently studying human papillomavirus. Some of the techniques could be used by individuals just moving into the area of human papillomavirus studies. However, to master most of these techniques would require additional demonstration or advice.
Features: This book covers a large variety of techniques starting with histochemical staining, isolation and detection of specific proteins, analysis of both DNA and protein markers associated with disease progression, study of oncogenesis and apoptosis and the use of cell culture systems to evaluate the human papilloma on human tissue.
Assessment: This is a useful book for individuals actively doing research on the human papillomaviruses. Because not all the techniques are unique to papillomavirus research, this volume would also be useful to researchers in other areas such as tumor progression, vaccine development, and host-pathogen interaction.

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