Quantification of resistance genes in veal manure by qPCR“But first we have to have some conditions clear”
Year: 2011
Type: dissertation
Abstract: The relation with the big therapeutic use of antibiotics in human and veterinary\nmedicine and the growing problem of resistance is getting more and more\nattention world wide. In the Netherlands, a political agreement has been reached\nto decrease antibiotic use in veal farming immediately. The aim of this research is\nto determine the qPCR detection limit of antibiotic resistance genes (tet(S)) in\nveal feces, to choose between two (absorbing overshoe and plastic cup) sampling\nmethods and to investigate PCR inhibition and its removal by dilution. Also a start\nwill be made in the setup of a protocol for quantification of Integrase genes\n(intI1) in feces samples. Integrons are intermediates in the pickup and expression\nof resistance genes and are often found on plasmids.\nThe sampling with a plastic cup will gave the best qPCR data. Because of\ninhibition factors, which are present in feces, a ten fold dilution of the DNA was\nnecessary for an optimal qPCR reaction. The detection limit for tet(S) was\ndetermined as 10^5 spiked genes per gram feces, which is a workable detection\nlimit for veal feces. Unstable standard dilutions were seen during this research,\nreasons and solutions for this are discussed.
Author: Roosmarijn E. C. Luiken
Language: English
Subfield: Pollution
Field: Environmental Science
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W176884499