Amplification of Salmonella chromosomal DNA using the polymerase chain reaction.
A Salmonella-specific polymerase chain reaction (PCR) was developed and standardized. The origin of the primers was a recombinant clone (C7) that contained Salmonella-specific HindIII fragment DNA of 2.1-kilobase pairs. Based on the sequence data of Salmonella enteritidis recombinant clone C7, two primers designated NK1 (21 nucleotides) and NK2 (24 nucleotides) were synthesized for use in the PCR. A Salmonella-specific 2.0-kilobase pair DNA product was amplified by the primers from 23 species of Salmonella, but not from 19 enteric and non-enteric bacteria. As little as 330 fg of Salmonella DNA was detected using either ethidium bromide/ultraviolet exposure of gels or Southern blot hybridization with a C7 clone.
Nguyen
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Department of Pathobiology, College of Agriculture and Natural Resources, University of Connecticut, Storrs 06269-3089.
Khan
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Lu
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D000818:Animals; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D015247:Deoxyribonuclease HindIII; D004755:Enterobacteriaceae / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry; D015183:Restriction Mapping; D012475:Salmonella / Q000235:genetics* / Q000302:isolation & purification; D013045:Species Specificity; D013552:Swine
D017931:DNA Primers; D004269:DNA, Bacterial; D015247:Deoxyribonuclease HindIII