PCR identification of Salmonella cells in food and stool samples after immunomagnetic separation.
The purpose of the present study was to investigate the application of various sample preparation methods (cell washing before lysis, purification of DNA using phenol extraction method, immunomagnetic separation-IMS) for the final PCR identification of Salmonella cells. The presence of PCR inhibitors in processed food products (milk powder and dried eggs) can be the cause of false-negative results in PCR without IMS of target cells. It was also demonstrated that IMS-PCR was successfully used for identification and quick confirmation of untypical Salmonella strains isolated from human stool samples and rabbit meat. However, IMS cannot eliminate intracellular PCR inhibitors present in immunoseparated Salmonella cells. These inhibitors must be taken into consideration in evaluation of PCR procedure.
Spanová
A
A
Department of Microbiology, Faculty of Science, Masaryk University, Brno, Czech Republic.
Rittich
B
B
Karpísková
R
R
Cechová
L
L
Skapová
D
D
D000818:Animals; D004531:Eggs / Q000382:microbiology; D005188:False Negative Reactions; D005243:Feces / Q000382:microbiology*; D005516:Food Microbiology; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008460:Meat / Q000382:microbiology; D008892:Milk / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011817:Rabbits; D012475:Salmonella / Q000235:genetics* / Q000302:isolation & purification*