Diagnosis of Sexually Transmitted Diseases Methods and Protocols

Sexually transmitted infections (STI) continue to be a major cause of morbidity and mortality, both in developed industrial countries as well as in the developing world. Human immunodeficiency virus infections and the ensuing opportunistic infections are a major drain on the human and financial resources of many countries in the developing world and even with the availability of effective treatment the epidemic is not yet contained. Screening in developing worlds is difficult and there is a need for simple reliable cheap diagnostic methods that can be performed at the point of care, on the same day and by staff with limited training. Molecular biological methods are very attractive for the diagnosis of STI since a well defined range of pathogens is responsible for the infection. Diagnosis of Sexually Transmitted Diseases: Methods and Protocols strives to cover the full range of molecular testing for STI. Chapters cover a variety of topics such as aspects of DNA extraction from small volume samples or difficult tissues, simple, nested or multiplex PCR, use of duplex primers or other modifications of primers and PCR conditions, sequence analysis for genotyping, denaturing gel analysis, microarrays using liquid beads or microspheres and silicon nanoparticle-enhanced microcantilever detection of DNA. Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Diagnosis of Sexually Transmitted Diseases: Methods and Protocols will be a valuable reference for the scientist looking to establish molecular methodologies for the diagnosis of STI tests in their own lab.

1. Overview of Molecular Biological Methods for the Detection of Pathogens Causing Sexually Transmitted Infections

                Fernando Vazquez, Santido Melón, Luis Otero, and María de Oña

2. Guidelines for the Qualitative Detection of Viral Genomes in Dried Blood Spots

                Davide Gibellini, Elisa De Crignis, and Maria Carla Re

3. Guidelines for the Quantification of HIV and HCV in Small Volume Whole Blood Samples

                Tony Bruns and Katrin Steinmetzer

4. Guidelines for High-Resolution Genotyping of Chlamydia trachomatis Using Multilocus Sequence Analysis

                Linus Christerson and Björn Herrmann

5. Guidelines for the Use of Molecular Tests for the Detection and Genotyping of Human Papilloma Virus from Clinical Specimens

                Sin Hang Lee

6. The Molecular Diagnosis of Sexually Transmitted Genital Ulcer Disease

                Cheng-Yen Chen and Ronald C. Ballard

7. Validation of a Sensitive and Specific Real-Time PCR for Detection and Quantitation of Hepatitis B Virus Covalently Closed Circular DNA in Plasma of Chronic Hepatitis B Patients

                R. B. Takkenberg, S. Menting, and M.G.H.M. Beld

8. Protocol for the Detection of Mycoplasma genitalium by PCR from Clinical Specimens and Subsequent Detection of Macrolide Resistance Mediating Mutations in Region V of the 23S rRNA Gene

                Jørgen Skov Jensen

9. Protocols for Detection and Typing of Treponema pallidum Using PCR Methods

                S.M. Bruisten

10. Protocol for the Extraction of DNA from Dried Blood in the Diagnosis of Convenital CMV Infection

                Jutte J.C. de Vries, Maria Barbi, Sandro Binda, and Eric C.J. Claas

11. Protocol for the Use of PCR-Denaturing Gradient Gel Electrophoresis and Quantitative PCR to Determine Vaginal Microflora Constitution and Pathogens in Bacterial Vaginosis

                Beatrice Vitali, Elena Biagi, and Patrizia Brigidi

12. Protocol for the Use of a Bead Array for the Multiple Detection of Genotype of Chlamydia trachomatis

                Chung-Ter uhang and Shu-Ying Li

13. Protocol for the Detection and Genotyping of Human Papilloma viruses Using a Liquid Bead Microarray Assay (LBMA)

                Stephen Cherne and Qinghua Feng

14. Protocol for the Use of Enzyme-Linked Hybridisation Assays for Genital Ulcer Disease

                Seweryn Bialasiewicz, Ian M. Mackay, David M. Whiley, and Theo P. Sloots

15. Protocol for the Rapid Detection of the Urogential Tract Mollicutes and Chlamydia with Concomitant LGV –(sub)typing

                Nadine Möbius, Wibke Brenneisen, Anke Schaeffer, and Birgit Henrich

16. Protocol for the Use of Self-Reporting Duplex Mutation Primers to Detect PCR Products in the Diagnosis of HBV

                Qian-Feng Xia

17. Protocol for the Use of a Rapid Real-Time PCR Method for the Detection of HIV-1 Proviral DNA Using Double-Stranded Primer

                Chou-Pong Pau, Susan K. Wells, and Timothy C. Granade

18. Protocol for the Use of Light Upon Extension (LUX) Real-Time PCR for the Determination of Viral Load in HBV Infection

                Guimin Li, Wangfeng Li, and Lixia Liu

19. Protocol for the Use of a Silica Nanoparticle-Enhanced Microcantilever Sensor-Based Method to Detect HBV at Femtomolar Concentrations

                Sang-Myung Lee, Kyo Seon Hwang, Sang Kyung Kim, and Tae Song Kim

20. Protocol for the Detection of Treponema pallidum in Paraffin-Embedded Specimens

Cheng-Yen Chen and Allan Pillay

21. Guidelines for the Use of Molecular Biological Methods to Detect Sexually Transmitted Pathogens in Cases of Suspected Sexual Abuse in Children

                Margaret R. Hammerschlag and Charlotte A. Gaydos

22. Protocol for the Molecular Detection of Antibiotic Resistance Mechanisms in Neisseria gonorrhoeae

                Namraj Goire, Theo P. Sloots, Michael D. Nissen, and David M. Whiley

23. Protocol for a Facile Multiplex PCR for Multi-Antimicrobial Resistance and Gonococcus Detection

                Ratana Lawung, Rungrot Cherdtrakulkiat, and Virapong Prachayasittikul

24. Protocol for Gene Expression Profiling Using DNA Microarrays in Neisseria gonorrhoeae

                Lydgia A. Jackson and David W. Dyer

25. Protocol for the Detection of Human Leukocyte Antigen Variants and Sexually Transmitted Infections: From Genotyping to Immunoassays

                Jianming Tang and Anju Bansal

26. Protocol for the Clonal Analysis of NK Cell Effector Functions by Multi-Parameter Flow Cytometry

                Kathrin Schönberg, Maryam Hejazi, and Markus Uhrberg

27. Protocol for a Mammalian Cell-Based Assay for Monitoring the HIV-1 Protease Activity

                Chitra Rajakuberan, Brett J. Hilton, and Roland Wolkowicz

28. Self Collection of Specimens for Nucleic Acid-Based Diagnosis of Pharyngeal, Cevicovaginal, Urethral and Rectal Neisseria gonorrhoeae and Chlamydia trachomatis Infections

                Vivian Levy, Craig S. Blackmore, and Jeffrey D. Klausner

29. Sexually Transmitted Diseases – Reflections on Metaphors and Ethics

                Gisela Badura-Lotter