Detection of Salmonella spp. in water using magnetic capture hybridization combined with PCR or real-time PCR

Authors
Thompson, D. E.; Rajal, Verónica Beatriz; De Batz, S.; Wuertz, S.
Publication Year
2006
Language
English
Format
article
Status
Published version
Description
The removal of target DNA by magnetic capture hybridization (MCH) from constituents inhibitory to amplification by polymerase chain reaction (PCR) was evaluated using Salmonella as the test pathogen. Hybrids were subjected to both conventional and quantitative real-time PCR (qPCR). When PCR inhibitors commonly found in water were added to the reaction, MCH-PCR increased the detection sensitivity on the order of 8 to 2,000-fold compared with the system using only PCR. To determine the selectivity of MCH for target DNA (Salmonella), different amounts of non-target DNA (Escherichia coli) were added to the qPCR reaction. The highest non-target DNA concentration interfered with the amplification by qPCR alone, while MCH-qPCR was unaffected. Average recovery of Salmonella DNA by MCH-qPCR was 31% using optimized buffers, washing solutions and enzymatic digestion. A recovery function was proposed in order to calculate the real cell number based on the measured value. Preliminary testing confirmed the suitability of this method for analysis of natural waters. © IWA Publishing 2006.
Fil: Thompson, D. E.. University of California at Davis; Estados Unidos
Fil: Rajal, Verónica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of California at Davis; Estados Unidos
Fil: De Batz, S.. University of California at Davis; Estados Unidos
Fil: Wuertz, S.. University of California at Davis; Estados Unidos
Subject
INHIBITORS
MAGNETIC BEADS
PCR
SALMONELLA
WATER
Oceanografía, Hidrología, Recursos Hídricos
Ciencias de la Tierra y relacionadas con el Medio Ambiente
CIENCIAS NATURALES Y EXACTAS
Access level
Open access
License
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repository
CONICET Digital (CONICET)
Institution
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identifier
oai:ri.conicet.gov.ar:11336/67164