Rapid detection of Salmonella in field samples by nested polymerase chain reaction.
A simple and universal protocol for the rapid detection of Salmonella spp. in various samples using nested polymerase chain reaction (PCR) was developed. The protocol takes advantage of the rapid purification and concentration of Salmonella by centrifugation onto a layer of 60% sucrose solution. DNA was released by treatment with proteinase K and Triton X-100. Even without pre-enrichment, the detection limit for the nested PCR was 10(5) CFU g-1 of faeces. After pre-enrichment, it was possible to detect Salmonella in faeces where their original concentration was approximately 10(2) CFU g-1 of faeces and in meat samples, it was possible to detect Salmonella in samples where their original concentration was less than 10 cells g-1 of minced meat.
Rychlík
I
I
Veterinary Research Institute, Hudcova, Brno, Czech Republic. rychlik@vri.cz
van Kesteren
L
L
Cardová
L
L
Svestková
A
A
Martínková
R
R
Sisák
F
F
D000818:Animals; D002645:Chickens / Q000382:microbiology*; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D005243:Feces / Q000382:microbiology*; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*
D003470:Culture Media; D004269:DNA, Bacterial