title: Innovative use of platinum compounds to selectively detect live microorganisms by polymerase chain reaction.
abstract: PCR cannot distinguish live microorganisms from dead ones. To circumvent this disadvantage, ethidium/propidium-monoazide (EMA/PMA) and psoralen to discriminate live from dead bacteria have been used for 2 decades. These methods require the use of numerous laborious procedures. We introduce an innovative method that uses platinum compounds, which are primarily used as catalysts in organic chemistry and partly used as anti-cancer drugs. Microorganisms are briefly exposed to platinum compounds in vivo, and these compounds penetrate dead (compromised) microorganisms but not live ones and are chelated by chromosomal DNA. The use of platinum compounds permits clear discrimination between live and dead microorganisms in water and milk (including Cronobacter sakazakii and Escherichia coli) via PCR compared with typically used PMA. This platinum-PCR method could enable the specific detection of viable coliforms in milk at a concentration of 5-10 CFU mL(-1) specified by EU/USA regulations after a 4-h process. For sample components, environmental water contains lower levels of PCR inhibitors than milk does, and milk is similar to infant formula, skim milk and blood; thus, the use of the platinum-PCR method could also prevent food poisoning due to the presence of C. sakazakii in dairy products. This method could provide outstanding rapidity for use in environmental/food/clinical tests. Platinum-PCR could also be a substitute for the typical culture-based methods currently used.
authors.affiliation: Functional Food Ingredients Department, Food Ingredients & Technology Institute. t_soezim@morinagamilk.co.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Soejima
authors.affiliation: Functional Food Ingredients Department, Food Ingredients & Technology Institute.
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Minami
authors.affiliation: Next Generation Science Institute, Morinaga Milk Industry Co., Ltd., 5-1-83, Higashihara, Zama City, Kanagawa Pref., 252-8583, Japan.
authors.forename: Jin-Zhong
authors.initials: JZ
authors.lastname: Xiao
authors.affiliation: Functional Food Ingredients Department, Food Ingredients & Technology Institute.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Abe
mesh_terms: D000818:Animals; D044083:Cronobacter sakazakii / Q000187:drug effects / Q000235:genetics; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D050296:Microbial Viability; D008892:Milk / Q000382:microbiology; D017671:Platinum Compounds / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014871:Water Microbiology
chemical_list: D004269:DNA, Bacterial; D017671:Platinum Compounds
keywords: bacteria; ethidium monoazide; pathogen; platinum; polymerase chain reaction; propidium monoazide