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Chapter 13 : Quantitative Molecular Methods

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Quantitative Molecular Methods, Page 1 of 2

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Abstract:

Diagnostic information describing the actual or relative density of specific viral nucleic acids in a human blood sample is most easily ascertained using quantitative molecular methods (aka viral loads), whose historical use spans more than two decades. A common function of quantitative viral load testing is to support clinical strategies and practices that monitor patients for human immunodeficiency virus (HIV) and most of the hepatitis viruses. For transplant patients, viral loads are monitored to detect numerous viruses classified in the herpes virus group, including cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpes virus 6 (HHV-6), among others. It seems that each year, there is a new application or interpretation for results of viral load assays. Bacterial load assays are uncommon; however, there is some discussion of its use for certain conditions.

Citation: Whitfield N, Wolk D. 2016. Quantitative Molecular Methods, p 145-166. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch13
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Figures

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FIGURE 1

(A) Fluorescence output from real-time PCR showing eight samples with corresponding copy numbers and corresponding standard curves derived from the values. Reactions occurring after cycle 45 could depict the formation and amplification of primer-dimers, so sequence analysis of these types of reaction may be necessary. (B) Example of a standard curve derived from real-time Q-PCR with calculations for PCR efficiency. The log of the starting copy number of nucleic acid is plotted against . Samples of unknown concentration can be assessed in comparison to the standard curve. Efficiency can be calculated from the slope of the regression line.

Citation: Whitfield N, Wolk D. 2016. Quantitative Molecular Methods, p 145-166. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch13
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Image of FIGURE 2
FIGURE 2

Citation: Whitfield N, Wolk D. 2016. Quantitative Molecular Methods, p 145-166. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch13
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FIGURE 3

Citation: Whitfield N, Wolk D. 2016. Quantitative Molecular Methods, p 145-166. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch13
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