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Pre-PCR processing in bioterrorism preparedness: improved diagnostic capabilities for laboratory response networks.

Hedman, Johannes LU ; Knutsson, Rickard ; Ansell, Ricky ; Rådström, Peter LU and Rasmusson, Birgitta LU (2013) In Biosecurity and Bioterrorism 11(Suppl 1). p.87-101
Abstract
Diagnostic DNA analysis using polymerase chain reaction (PCR) has become a valuable tool for rapid detection of biothreat agents. However, analysis is often challenging because of the limited size, quality, and purity of the biological target. Pre-PCR processing is an integrated concept in which the issues of analytical limit of detection and simplicity for automation are addressed in all steps leading up to PCR amplification--that is, sampling, sample treatment, and the chemical composition of PCR. The sampling method should maximize target uptake and minimize uptake of extraneous substances that could impair the analysis--so-called PCR inhibitors. In sample treatment, there is a trade-off between yield and purity, as extensive... (More)
Diagnostic DNA analysis using polymerase chain reaction (PCR) has become a valuable tool for rapid detection of biothreat agents. However, analysis is often challenging because of the limited size, quality, and purity of the biological target. Pre-PCR processing is an integrated concept in which the issues of analytical limit of detection and simplicity for automation are addressed in all steps leading up to PCR amplification--that is, sampling, sample treatment, and the chemical composition of PCR. The sampling method should maximize target uptake and minimize uptake of extraneous substances that could impair the analysis--so-called PCR inhibitors. In sample treatment, there is a trade-off between yield and purity, as extensive purification leads to DNA loss. A cornerstone of pre-PCR processing is to apply DNA polymerase-buffer systems that are tolerant to specific sample impurities, thereby lowering the need for expensive purification steps and maximizing DNA recovery. Improved awareness among Laboratory Response Networks (LRNs) regarding pre-PCR processing is important, as ineffective sample processing leads to increased cost and possibly false-negative or ambiguous results, hindering the decision-making process in a bioterrorism crisis. This article covers the nature and mechanisms of PCR-inhibitory substances relevant for agroterrorism and bioterrorism preparedness, methods for quality control of PCR reactions, and applications of pre-PCR processing to optimize and simplify the analysis of various biothreat agents. Knowledge about pre-PCR processing will improve diagnostic capabilities of LRNs involved in the response to bioterrorism incidents. (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biosecurity and Bioterrorism
volume
11
issue
Suppl 1
pages
87 - 101
publisher
Mary Ann Liebert, Inc.
external identifiers
  • pmid:23971826
  • wos:000336524100011
  • scopus:84883259306
  • pmid:23971826
ISSN
1557-850X
DOI
10.1089/bsp.2012.0090
language
English
LU publication?
yes
id
c51cec08-d761-4cf6-97db-4b5e145cc567 (old id 4179792)
date added to LUP
2016-04-01 09:57:23
date last changed
2022-04-12 00:35:51
@article{c51cec08-d761-4cf6-97db-4b5e145cc567,
  abstract     = {{Diagnostic DNA analysis using polymerase chain reaction (PCR) has become a valuable tool for rapid detection of biothreat agents. However, analysis is often challenging because of the limited size, quality, and purity of the biological target. Pre-PCR processing is an integrated concept in which the issues of analytical limit of detection and simplicity for automation are addressed in all steps leading up to PCR amplification--that is, sampling, sample treatment, and the chemical composition of PCR. The sampling method should maximize target uptake and minimize uptake of extraneous substances that could impair the analysis--so-called PCR inhibitors. In sample treatment, there is a trade-off between yield and purity, as extensive purification leads to DNA loss. A cornerstone of pre-PCR processing is to apply DNA polymerase-buffer systems that are tolerant to specific sample impurities, thereby lowering the need for expensive purification steps and maximizing DNA recovery. Improved awareness among Laboratory Response Networks (LRNs) regarding pre-PCR processing is important, as ineffective sample processing leads to increased cost and possibly false-negative or ambiguous results, hindering the decision-making process in a bioterrorism crisis. This article covers the nature and mechanisms of PCR-inhibitory substances relevant for agroterrorism and bioterrorism preparedness, methods for quality control of PCR reactions, and applications of pre-PCR processing to optimize and simplify the analysis of various biothreat agents. Knowledge about pre-PCR processing will improve diagnostic capabilities of LRNs involved in the response to bioterrorism incidents.}},
  author       = {{Hedman, Johannes and Knutsson, Rickard and Ansell, Ricky and Rådström, Peter and Rasmusson, Birgitta}},
  issn         = {{1557-850X}},
  language     = {{eng}},
  number       = {{Suppl 1}},
  pages        = {{87--101}},
  publisher    = {{Mary Ann Liebert, Inc.}},
  series       = {{Biosecurity and Bioterrorism}},
  title        = {{Pre-PCR processing in bioterrorism preparedness: improved diagnostic capabilities for laboratory response networks.}},
  url          = {{http://dx.doi.org/10.1089/bsp.2012.0090}},
  doi          = {{10.1089/bsp.2012.0090}},
  volume       = {{11}},
  year         = {{2013}},
}