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Use of aqueous two-phase systems in sample preparation for polymerase chain reaction-based detection of microorganisms

Lantz, Pär-Gunnar; Tjerneld, Folke LU ; Hahn-Hägerdal, Bärbel LU and Rådström, Peter LU (1996) In Journal of Chromatography. B 680(1-2). p.165-170
Abstract
An aqueous two-phase system, consisting of poly(ethylene glycol) (PEG) and dextran, was employed to separate polymerase chain reaction (PCR)-inhibitory substances from bacterial cells. The PCR inhibition of four soft cheeses was examined and three of them were found to be strongly PCR-inhibitory. Extraction of the PCR-inhibitory soft cheeses inoculated with Listeria monocytogenes in an aqueous two-phase system containing 8% (w/w) PEG 4000 and 8% (w/w) dextran 500, was found to lower the PCR detection level of L. monocytogenes by more than four orders of magnitude in two of the cheeses compared to the case where no such sample pretreatment was performed. Depending on the type of cheese used, the PCR-inhibitory factors were found to be... (More)
An aqueous two-phase system, consisting of poly(ethylene glycol) (PEG) and dextran, was employed to separate polymerase chain reaction (PCR)-inhibitory substances from bacterial cells. The PCR inhibition of four soft cheeses was examined and three of them were found to be strongly PCR-inhibitory. Extraction of the PCR-inhibitory soft cheeses inoculated with Listeria monocytogenes in an aqueous two-phase system containing 8% (w/w) PEG 4000 and 8% (w/w) dextran 500, was found to lower the PCR detection level of L. monocytogenes by more than four orders of magnitude in two of the cheeses compared to the case where no such sample pretreatment was performed. Depending on the type of cheese used, the PCR-inhibitory factors were found to be enriched in either the top or bottom phase in the aqueous two-phase system. These results show that different soft cheeses contain different types and amounts of PCR-inhibitory substances. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
PCR-inhibitory factors, Sample preparation method, aqueous, Two-phase systems, Cheese, Polymerase chain reaction, Microorganisms, Aqueous two-phase systems, Listeria monocytogenes
in
Journal of Chromatography. B
volume
680
issue
1-2
pages
165 - 170
publisher
Elsevier
external identifiers
  • scopus:0029954427
ISSN
1873-376X
DOI
10.1016/0378-4347(95)00496-3
language
English
LU publication?
yes
id
a2b38837-1e78-46f3-a126-43b85480d953 (old id 126294)
date added to LUP
2007-06-28 15:50:19
date last changed
2017-01-01 05:14:04
@article{a2b38837-1e78-46f3-a126-43b85480d953,
  abstract     = {An aqueous two-phase system, consisting of poly(ethylene glycol) (PEG) and dextran, was employed to separate polymerase chain reaction (PCR)-inhibitory substances from bacterial cells. The PCR inhibition of four soft cheeses was examined and three of them were found to be strongly PCR-inhibitory. Extraction of the PCR-inhibitory soft cheeses inoculated with Listeria monocytogenes in an aqueous two-phase system containing 8% (w/w) PEG 4000 and 8% (w/w) dextran 500, was found to lower the PCR detection level of L. monocytogenes by more than four orders of magnitude in two of the cheeses compared to the case where no such sample pretreatment was performed. Depending on the type of cheese used, the PCR-inhibitory factors were found to be enriched in either the top or bottom phase in the aqueous two-phase system. These results show that different soft cheeses contain different types and amounts of PCR-inhibitory substances.},
  author       = {Lantz, Pär-Gunnar and Tjerneld, Folke and Hahn-Hägerdal, Bärbel and Rådström, Peter},
  issn         = {1873-376X},
  keyword      = {PCR-inhibitory factors,Sample preparation method,aqueous,Two-phase systems,Cheese,Polymerase chain reaction,Microorganisms,Aqueous two-phase systems,Listeria monocytogenes},
  language     = {eng},
  number       = {1-2},
  pages        = {165--170},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography. B},
  title        = {Use of aqueous two-phase systems in sample preparation for polymerase chain reaction-based detection of microorganisms},
  url          = {http://dx.doi.org/10.1016/0378-4347(95)00496-3},
  volume       = {680},
  year         = {1996},
}