Rapid Detection of Staphylococcus aureus in Aquatic Products by Duplex PCR-DHPLC

doi:10.7506/spkx1002-6630-201306044

Abstract

Abstract: Staphylococcus aureus is an important pathogen that can cause food poisoning. Two pairs of specific primers were designed according to the sequences of the sea gene (staphylococcal enterotoxin A) and the nuc gene from Staphylococcus aureus for the development of a duplex polymerase chain reaction-denaturing high performance liquid chromatography (PCRDHPLC) to detect Staphylococcus aureus strains in aquatic products. The positive and negative detection using this duplex PCR-DHPLC method was observed in 8 and 22 out of 30 strains, respectively. This method had high sensitivity with the detection limit of 50 CFU/mL, which revealed an improvement by 10 fold compared with the ordinary gel electrophoresis. Based on the duplex PCR-DHPLC method, 240 aquatic products were analyzed and showed a positive detection rate of 9.2% for Staphylococcus aureus contamination, which did not exhibit significant difference in positive detection rate from national standard approach. Therefore, duplex PCR-DHPLC can be used for specific, sensitive, rapid and high throughput detection of Staphylococcus aureus in aquatic products, and also can be used to determine the strength of pathogenicity of Staphylococcus aureus through analysis of the staphylococcal enterotoxin gene.

Key words: Staphylococcus aureus, duplex PCR-DHPLC, aquatic products, detection

CLC Number:

R155.5

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