In house validation of real-time PCR method for detection of Clostridium botulinum in food and feed matrixes

• Autorzy: Grenda T. , Kwiatek K.
• Języki publikacji: EN

Abstrakty

EN

For validation purposes, characteristic parameters for quantitative detection were estimated according to the PN-EN ISO 16140. Additionally, a comparison between validated real-time PCR method and traditional methods based on the isolation of this pathogen on differential agar media was conducted. The validated method has shown the possibility of detection of 58 copies of ntnh gene (genome equivalents) for DNA obtained from dilution of pure Clostridium botulinum NCTC 887 cells. For DNA obtained from the contaminated food and feed samples inhibitory effect was observed. The tested method has shown high specificity proved by the examination of DNA obtained from C. botulinum reference strains and other strains of Clostridium sp. The specificity has also proved the obtained concordance between results from analyses using test on laboratory mice with those from analyses using the tested real-time PCR method. The obtained results have shown that the described method gives the possibility to detect the pathogen without isolation and to shorten time of analysis in comparison to the traditional methods, based on isolation of this pathogen on differential agar media.

Słowa kluczowe

EN

validation; real-time polymerase chain reaction; detection; Clostridium botulinum; food; feed; bacteria

• Wydawca: -
• Czasopismo: Bulletin of the Veterinary Institute in Puławy
• Rocznik: 2010
• Tom: 54
• Numer: 4
• Opis fizyczny: p.557-562,fig.,ref.

Twórcy

autor

Grenda T.

• Department of Hygiene of Feeding Stuffs, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Kwiatek K.

Bibliografia

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