title: Use of oligonucleotide array for identification of six foodborne pathogens and Pseudomonas aeruginosa grown on selective media.
abstract: Identification of presumptive foodborne pathogens grown on selective media may take one to several days and requires a different battery of biochemical tests for each microorganism. A molecular identification method was developed in which universal primers were used to amplify the 16S to 23S rDNA intergenic spacer of target microorganisms, and PCR products were hybridized to a panel of species-specific oligonucleotides that were immobilized on a nylon membrane. The seven target microorganisms were Bacillus cereus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella, Staphylococcus aureus, and Vibrio parahaemolyticus. After testing a large collection of target bacteria (29 to 51 strains) and nontarget bacteria (> 500 strains), the performances (sensitivity and specificity) of the oligonucleotide array were as follows: B. cereus (100 and 77%), E. coli (100 and 100%), L. monocytogenes (100 and 90%), P. aeruginosa (100 and 100%), Salmonella (100 and 100%), S. aureus (100 and 100%), and V. parahaemolyticus (100 and 94.2%). Other species in the B. cereus group cross-hybridized to the probes used for identification of B. cereus, and positive results should be confirmed by additional morphological observation of colonies. Listeria innocua cross-reacted with probes used to identify L. monocytogenes, but a simple hemolysis test was used to differentiate the two species. Some strains of Vibrio harveyi and Vibrio mimicus cross-hybridized with probes used for identification of V. parahaemolyticus and caused false-positive reactions. The advantage of the array is that a common protocol was used to identify the seven target microorganisms and multiple different microorganisms could be simultaneously identified on a single array.
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, School of Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China.
authors.forename: Miao Chu
authors.initials: MC
authors.lastname: Lin
authors.forename: Ay Huey
authors.initials: AH
authors.lastname: Huang
authors.forename: Hau Yang
authors.initials: HY
authors.lastname: Tsen
authors.forename: Hin-Chung
authors.initials: HC
authors.lastname: Wong
authors.forename: Tsung Chain
authors.initials: TC
authors.lastname: Chang
mesh_terms: D001409:Bacillus cereus / Q000235:genetics / Q000302:isolation & purification; D015169:Colony Count, Microbial; D003470:Culture Media / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D005189:False Positive Reactions; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000302:isolation & purification
chemical_list: D003470:Culture Media