Pre-PCR processing in bioterrorism preparedness: improved diagnostic capabilities for laboratory response networks.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4179792
- author
- Hedman, Johannes LU ; Knutsson, Rickard ; Ansell, Ricky ; Rådström, Peter LU and Rasmusson, Birgitta LU
- organization
- publishing date
- 2013
- 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}}, }