Development of PCR primers and a DNA macroarray for the simultaneous detection of major staphylococcus species using groESL gene

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Title:	Development of PCR primers and a DNA macroarray for the simultaneous detection of major staphylococcus species using groESL gene
Authors:	Chiang, Y.-C., Lu, H.-C., Li, S.-C., Chang, Y.-H., Chen, H.-Y., Lin, C.-W., Tsen, H.-Y.
Contributors:	Department of Food Science, Tunghai University
Keywords:	article;bacteremia;bacterial gene;bacterium detection;bacterium identification;consensus sequence;DNA determination;DNA probe;food poisoning;groESL gene;nonhuman;nucleotide sequence;polymerase chain reaction;priority journal;species identification;Staphylococcus;Staphylococcus aureus;Staphylococcus carnosus;Staphylococcus haemolyticus;Staphylococcus hyicus;Staphylococcus intermedius;Staphylococcus saprophyticus;Staphylococcus xylosus
Date:	2012
Issue Date:	2013-06-11T09:04:22Z (UTC)
Abstract:	Staphylococcus spp., including S. aureus, S. intermedius, S. hyicus, S. epidermidis, S. saprophyticus, S. haemolyticus, S. xylosus, and S. carnosus, are major bacterial species associated with food poisoning, and human and veterinary clinics. Traditional methods for the identification of these staphylococci are time-consuming, laborious, or inaccurate. Therefore, rapid and accurate diagnostic methods are needed. In this study, we designed the DNA probes and polymerase chain reaction (PCR) primers for the detection of the aforementioned Staphylococcus species. These primers were proved to be specific for the detection of their corresponding target strains. Furthermore, by using a consensus primer pair, we were able to co-amplify the intergenic region of groES-groEL for these staphylococci. Followed by a chromogenic macroarray system with the specific probes on the plastic chips, these staphylococci in milk products or clinical samples could be simultaneously detected. When the system was used for the inspection of milk or urine samples containing N?10 0 target cells per milliliter of the sample, all these staphylococcal species could be identified after an 8-h pre-enrichment step. This system also allowed the adequate diagnosis of bacteremia, since N?10 0 target cells per milliliter of the blood samples could be detected after a 12-h pre-enrichment. Compared to the multiplex PCR method, this approach has the additional advantage that it allowed the discrimination of more bacterial strains-even some bacterial strains that may generate PCR products with the same molecular sizes. ? 2012, Mary Ann Liebert, Inc.
Relation:	Foodborne Pathogens and Disease 9 (3) , pp. 249-257
Appears in Collections:	[食品科學系所] 期刊論文

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