Inhibition of PCR by components of food samples, microbial diagnostic assays and DNA-extraction solutions.
We have tested the influence on the polymerase chain reaction (PCR) of a large number of compounds found in food, in media used for selective propagation of food-borne pathogens or in DNA-extraction methods. PCR was found to be sensitive to large volumes of complex food samples containing high amounts of fat and protein, however, an extraction procedure based on treatment with hot NaOH/SDS reduced the effect significantly. Some culture media (Fraser, MLEB, MRB and Rappaport) interfered with the analysis and for most of the media it was possible to assign the inhibitory effect to one or more individual components. Several compounds (detergents, lysozyme, NaOH, alcohols, EDGA, EGTA) used in DNA extraction procedures were found to have some inhibitory effect. The inhibitory effects need to be taken into consideration when designing new tests.
Rossen
L
L
Department of Molecular Food Technology, Biotechnological Institute, Lyngby, Denmark.
Nørskov
P
P
Holmstrøm
K
K
Rasmussen
O F
OF
D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D002611:Cheese; D002645:Chickens; D003470:Culture Media; D004247:DNA / Q000032:analysis; D005516:Food Microbiology; D008460:Meat; D008461:Meat Products; D016133:Polymerase Chain Reaction; D013552:Swine
D003470:Culture Media; D004247:DNA