title: Detection of Alicyclobacillus species in fruit juice using a random genomic DNA microarray chip.
abstract: This study describes a method using a DNA microarray chip to rapidly and simultaneously detect Alicyclobacillus species in orange juice based on the hybridization of genomic DNA with random probes. Three food spoilage bacteria were used in this study: Alicyclobacillus acidocaldarius, Alicyclobacillus acidoterrestris, and Alicyclobacillus cycloheptanicus. The three Alicyclobacillus species were adjusted to 2 × 10(3) CFU/ml and inoculated into pasteurized 100% pure orange juice. Cy5-dCTP labeling was used for reference signals, and Cy3-dCTP was labeled for target genomic DNA. The molar ratio of 1:1 of Cy3-dCTP and Cy5-dCTP was used. DNA microarray chips were fabricated using randomly fragmented DNA of Alicyclobacillus spp. and were hybridized with genomic DNA extracted from Bacillus spp. Genomic DNA extracted from Alicyclobacillus spp. showed a significantly higher hybridization rate compared with DNA of Bacillus spp., thereby distinguishing Alicyclobacillus spp. from Bacillus spp. The results showed that the microarray DNA chip containing randomly fragmented genomic DNA was specific and clearly identified specific food spoilage bacteria. This microarray system is a good tool for rapid and specific detection of thermophilic spoilage bacteria, mainly Alicyclobacillus spp., and is useful and applicable to the fruit juice industry.
authors.affiliation: College of Life Sciences and Biotechnology, Korea University, 5-1 Anam-dong, Seongbuk-gu, Seoul 136-701, Republic of Korea.
authors.forename: Jun Hyeong
authors.initials: JH
authors.lastname: Jang
authors.forename: Sun-Joong
authors.initials: SJ
authors.lastname: Kim
authors.forename: Bo Hyun
authors.initials: BH
authors.lastname: Yoon
authors.forename: Jee-Hoon
authors.initials: JH
authors.lastname: Ryu
authors.forename: Man Bock
authors.initials: MB
authors.lastname: Gu
authors.forename: Hyo-Ihl
authors.initials: HI
authors.lastname: Chang
mesh_terms: D056450:Alicyclobacillus / Q000302:isolation & purification*; D001628:Beverages / Q000382:microbiology*; D002232:Carbocyanines; D032084:Citrus sinensis; D003257:Consumer Product Safety; D004269:DNA, Bacterial / Q000032:analysis*; D003845:Deoxycytosine Nucleotides; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D006801:Humans; D046228:Microarray Analysis; D013045:Species Specificity
chemical_list: D002232:Carbocyanines; C544355:Cy5-dCTP; D004269:DNA, Bacterial; D003845:Deoxycytosine Nucleotides