PMID: 9659542Jul 11, 1998Paper

Detection of Staphylococcus aureus in milk by use of polymerase chain reaction analysis

American Journal of Veterinary Research
M A KhanB K Baek

Abstract

To evaluate polymerase chain reaction (PCR) analysis for detection of Staphylococcus aureus (nuc gene) in fresh and formalin-preserved milk. Samples from 80 lactating sheep and 100 lactating dairy cows. 4 lactating sheep were inoculated with S aureus by intramammary infusion. A set of primers specific for the nuc gene of S aureus was used to develop a PCR technique, and modification of the rapid boil method was used to isolate bacterial DNA. Milk was obtained from experimentally infected sheep before and after infusion with S aureus, and from the 100 cows and remaining 76 sheep. Samples were screened by bacteriologic culture and PCR. To validate the PCR assay, S aureus or other pathogens were added to distilled water and "normal" sheep milk samples, with and without formalin. The PCR assay was 100% specific for S aureus when known negative and positive samples were tested. Sensitivity was 100% for samples with added S aureus or other pathogens. Sensitivity was lower for samples obtained from experimentally infected sheep, but increased from 53% to 90% with increased washing of target DNA. The PCR technique based on the nuc gene is able to detect S aureus in sheep milk yields results faster than does traditional culturing, is hi...Continue Reading

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