title: Evaluation of an improved bioluminescence assay for the detection of bacteria in soy milk.
abstract: Because soy milk is nutrient rich and nearly neutral in pH, it favors the growth of microbial contaminants. To ensure that soy milk meets food-safety standards, it must be pasteurized and have its sterility confirmed. ATP bioluminescence assay has become a widely accepted means of detecting food microorganisms. However, the high background bioluminescence intensity of soy milk has rendered it unsuitable for ATP analysis. Here, we tested the efficacy of an improved pre-treated bioluminescence assay on soy milk. By comparing background bioluminescence intensities obtained by the conventional and improved methods, we demonstrated that our method significantly reduces soy milk background bioluminescence. The dose-response curve of the assay was tested with serial dilutions of Bacillus sp. culture. An extremely strong log-linear relation between the bioluminescence intensity relative light units and colony formation units CFU/ml emerged for the tested strain. The detection limit of the assay was estimated as 5.2×10(3) CFU/ml from the dose-response curve and an imposed signal limit was three times the background level. The results showed that contaminated samples could be easily detected within 24 h using our improved bioluminescence assay.
authors.affiliation: Research and Development Division, Kikkoman Corporation, Noda, Chiba, Japan. yshinozaki@mail.kikkoman.co.jp
authors.forename: Yohei
authors.initials: Y
authors.lastname: Shinozaki
authors.forename: Jun
authors.initials: J
authors.lastname: Sato
authors.forename: Toshinori
authors.initials: T
authors.lastname: Igarashi
authors.forename: Shigeya
authors.initials: S
authors.lastname: Suzuki
authors.forename: Kazunori
authors.initials: K
authors.lastname: Nishimoto
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Harada
mesh_terms: D001419:Bacteria / Q000254:growth & development / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D005516:Food Microbiology; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D041942:Soy Milk; D013171:Spores, Bacterial