Title of article :
Direct detection and genotyping of Toxoplasma gondii in meat samples using magnetic capture and PCR
Author/Authors :
Opsteegh، نويسنده , , Marieke and Langelaar، نويسنده , , Merel and Sprong، نويسنده , , Hein and den Hartog، نويسنده , , Laurien and De Craeye، نويسنده , , Stéphane and Bokken، نويسنده , , Gertie and Ajzenberg، نويسنده , , Daniel and Kijlstra، نويسنده , , Aize and der Giessen، نويسنده , , Joke van، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2010
Pages :
9
From page :
193
To page :
201
Abstract :
Different transmission routes, including the ingestion of undercooked meat, can result in Toxoplasma gondii infection in humans. The development of effective prevention strategies is hampered by a lack of quantitative information on the contamination level of different types of meat. Therefore, we developed a method for detection and quantification of T. gondii. The method involved preparation of crude DNA extract from hundred gram samples of meat, magnetic capture of T. gondii DNA and, quantitative real-time PCR targeting the T. gondii 529-bp repeat element. The detection limit of this assay was approximately 230 tachyzoites per 100 g of meat sample. There was a linear relation between the number of parasites added to the samples and Cp-values. Results obtained with the PCR method were comparable to bioassay results for experimentally infected pigs, and to serological findings for sheep. In addition, the T. gondii in 50% of the positive sheep samples could be genotyped by sequencing of the GRA6 gene, after isolation of the gene by magnetic capture. Two subtypes of GRA6 type II were identified in the 16 samples from sheep. For seven samples, the identification of T. gondii as type II was confirmed by microsatellite typing. The PCR method can be used as an alternative to bioassay for detection and genotyping of T. gondii, and to quantify the organism in meat samples of various sources.
Keywords :
quantitative PCR , Genotyping , Source attribution , bioassay , TOXOPLASMA GONDII , Meat , magnetic capture , detection
Journal title :
International Journal of Food Microbiology
Serial Year :
2010
Journal title :
International Journal of Food Microbiology
Record number :
2115566
Link To Document :
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