Quantification of Salmonella from pig faeces by real-time PCR
2006
Malorny,B. | Szabo,I. | Fröder,H. | Nöckler,K. | Bunge,C. | Helmuth,R.
allemand. In order to assess the hygiene status and provide appropriate risk management options it is a prerequisite to estimate the levels of Salmonella contamination in a swineherd or at the slaughterhouse. Currently culture-based methods are used (Most Probable Number, direct plating) to investigate the level of contamination mainly from faeces . However, high levels of background flora can disturb the growth of Salmonella and high numbers of samples to investigate is time and labor intensive. The use of real-time PCR is a good alternative method for quantification of Salmonella in pig faeces. It is fast and can be automated.We have developed a protocol based on real-time PCR to quantify Salmonella DNA in pig faeces. Salmonella DNA is directly purified from the faecal material with the the QIAmp Stool kit (Qiagen). A preliminary "in-house" validation study, using faeces obtained over a four week period from pigs which have been infected with a nalidixic acid resistant Salmonella Typhimurium DT104 strain, showed a good agreement between the real-time PCR based method and direct plating on XLD agar containing 50 µg/ml nalidixic acid. Accurate quantification is possible over a five log linear range down to 100-200 CFU/g of faeces. This real-time PCR based method is a new tool to provide easier quantitative data for risk assessment models.
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