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Chapter 10 : Nucleic Acid Extraction in Diagnostic Virology

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Nucleic Acid Extraction in Diagnostic Virology, Page 1 of 2

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Abstract:

Nucleic acid (NA) extraction is a critical step used in molecular biology and molecular diagnostics (1–5). Successful extraction of NA depends on the quantitative recovery of pure molecules of RNA and DNA in an undegraded form. Salts, for example, are common impurities in NA samples, and it is important that they are removed from NA before any downstream processes and analyses can be performed (1). Therefore, single or multiple separation and/or purification steps are needed to desalt the sample containing the NA. The process of extraction and purification of NA also removes a variety of inhibitors of downstream NA amplification procedures. The first step of NA extraction and purification involves cell lysis to liberate NA from cell nuclei or pathogens. Effective NA extraction methods include reagents that inactivate nucleases (DNase and RNase) to preserve the NA in an intact state. The final steps involve separation and recovery of the NA free of cellular debris, proteins, and various potential inhibitors of downstream assays (1–5).

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
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Figures

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FIGURE 1

TruTip Technology. (a) TruTip Design (b) TruTip Extraction Process.

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
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Image of FIGURE 2
FIGURE 2

Reaction strip from the BD MAX Instrument. (a) Current configuration: one extraction and one polymerase chain reaction (PCR) amplification. (b) Future configuration: one extraction and two PCR amplifications.

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
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Tables

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TABLE 1

Instruments that automate the column extraction process

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
Generic image for table
TABLE 2

Instruments that use variations of magnetic particle capture for nucleic acid extraction

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
Generic image for table
TABLE 3

Automated extraction systems with PCR mix setup function

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
Generic image for table
TABLE 4

Systems that provide total automation with sample-in, result-out functionality

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10
Generic image for table
TABLE 5

Fully automated sample-to-answer instruments in development

Citation: Widen R, Silbert S. 2016. Nucleic Acid Extraction in Diagnostic Virology, p 117-128. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch10

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