By Robert K. Hamatake, Johnson Y. N. Lau
A broad-ranging choice of middle ideas for the research of HBV and HDV infections and for the improvement of remedies to regard them. during this first of 2 volumes Detection, Genotypes, and Characterization, the authors specialize in with ease reproducible molecular equipment for the identity and quantification of viral markers, the detection and impression of viral versions, and the examine of the viral lifestyles cycle. each one totally established protocol is defined in step by step element by means of a longtime professional within the box and contains a history creation outlining the primary underlying the process, gear and reagent lists, and pointers on troubleshooting and averting identified pitfalls. An accompanying moment quantity, Immunology, version platforms, and medical stories, includes trouble-free protocols for the research of host immune responses to an infection, in vitro and in vivo types of an infection, and the advance of antivirals. For either volumes: A broad-ranging choice of middle innovations for the examine of HBV and HDV infections and for the improvement of treatments to regard them. the 1st quantity Detection, Genotypes, and Characterization, the authors concentrate on with no trouble reproducible molecular equipment for the identity and quantification of viral markers, the detection and effect of viral editions, and the learn of the viral lifestyles cycle. the second one quantity, Immunology, version structures, and medical stories, includes simple protocols for the examine of host immune responses to an infection, in vitro and in vivo types of an infection, and the improvement of antivirals. each one absolutely demonstrated protocol is defined in step by step element through a longtime professional within the box and encompasses a history advent outlining the primary underlying the procedure, gear and reagent lists, and tips about troubleshooting and heading off recognized pitfalls. The set deals either new and skilled investigators an encyclopedic choice of strong instruments for learning HBV and HDV infections, a vital source for locating new treatments to regard chronically contaminated sufferers.
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Extra resources for Hepatitis B and D Protocols: Volume 1: Detection, Genotypes, and Characterization
However, the typical range of HBsAg concentration is between 1000 and 100,000 ng/mL. Thus, reliable quantitation by ELISA requires dilution series. Furthermore, ELISAs are usually not calibrated in quantitative units. 4. Deﬁnition of HBsAg Units There is much confusion about units of HBsAg reactivity. Many workers avoid quantitation completely or provide sample to cutoff ratio (S/CO) units. Occasionally, these units are converted to ng HBsAg, but most communicated ng/mL values are too high, because the HBsAg used for calibration had partially lost reactivity.
4. These steps should be performed in the extraction area. 5. These steps should be performed in the ampliﬁcation area. Acknowledgments The method described here evolved from informal discussions with several interested parties. Carolyn Luscombe and Madhu Singh from VIRIDAE (Vancouver, Canada) made important contributions in the early phase of development. Craig Gibbs from Gilead Sciences (Foster City, CA) provided substantial funding, which allowed the assays to progress from a research test to a research tool, and encouraged participation and contributions of other scientists.
24. Check the intensity of positive staining under an inverted microscope. Do not allow the sections to dry (see Note 5). 25. Rinse the sections with tap water. 26. 1% nuclear fast red solution for 5 min. 27. Brieﬂy rinse the sections with tap water to remove excess nuclear fast red solution. 28. Dehydrate the sections through a standard ethanol series, clear through xylene and mount the slides (see Note 6). 29. Remember that controls are critical for this experiment (see Note 7). 30. Results from one of the experiments are illustrated in Fig 1.
Hepatitis B and D Protocols: Volume 1: Detection, Genotypes, and Characterization by Robert K. Hamatake, Johnson Y. N. Lau