Diagnostic Virology Protocols (Methods in Molecular by John R. Stephenson, Alan Warnes

By John R. Stephenson, Alan Warnes

A set of state of the art options for detecting many of the significant viruses that afflict mankind, together with influenza, hepatitis, herpes, polio, mumps, HIV, and lots of extra. The innovations are well-tested, simply reproducible, and comfortably hire the entire new technologies-PCR, RIA, ELISA, and latex-agglutination-that have revolutionized the sphere. those equipment not just give the chance to do the required research in hours rather than days, yet is additionally automatic in a laboratory havng in simple terms low degrees of organic containment. often, the protocols for viruses inflicting human illnesses could be tailored to comparable viruses of veterinary value. via its state of the art tools a doctor can, for the 1st time, make certain early in a viral an infection which antiviral drug could be used and reduce the interval of remedy to prevent pointless unwanted side effects.

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42 Muir 11. Tissue grinding: To minimize endogenous RNase activity, it is important to keep tissue frozen during the grinding procedure Prechilling scalpel blades and mortars and pestles to -70°C 1sessential for this. 12 Chilling the tissue portion m liquid nitrogen facilitates grmdmg by making the tissue brittle 13. l-mL mmihomogemzers are used to mmlmize sample loss, homogenization is more effective when the full 400 pL of RNAzol B 1s added to the homogemzer. Homogemzatlon m smaller volumes of RNAzol B results in excessive foaming.

9. Crushed wet me. 10. RNAzol B. 3. Preparation 1. 2 3. 4 5 6. 7. 8. 9 10. 11. 12 13. of Paraffin-Embedded Tissue Mtcrotome employing dtsposable knife blades. Sterile, disposable plastic forceps. Filtered 2-20-, 2%200-, and 100-l 000-pL pipet trps. Sterile, nuclease/protease free 0 65-mL Eppendorf tubes Programmable thermal cycler. n-Octane (Sigma o-2001) Absolute ethanol. 0 05MEDTA Tween-20 (Sigma P-9416). Proteinase K (Sigma P-49 14). Sterile molecular btology grade water. 4. Reverse Transcription-PCR 1 2.

Eds ), American Society for Microbiology, Washington, DC, pp. 6066 18. 3. Monath, T. , Nystrom, R. , Bailey, R. , Calisher, C. , and Muth, D. J. (1984) Immunoglobulm M antibody capture enzyme-linked nnmunosorbent assay for diagnosis of St Louis encephalitis. J Clm Muzroblol 20,784-790. 4. , and Bartlett, A. ), American Society for Microbiology, Washmgton, DC, pp. 506-5 12. 5. Roehrig, J. , Hunt, A. , Tsar, T F , and Trent, D. W. (1990) Identification of monoclonal anttbodies capable of differentiating antigemc varieties of eastern equine encephahtis viruses.

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