title: An integrated microfluidic chip for the detection of bacteria - A proof of concept.
abstract: We designed a microfluidic chip as a proof of concept for the detection of bacterial DNA. The chip was fabricated with poly-dimethylsiloxane (PDMS). It included a solid phase extraction (SPE) chamber, two separate channels and multiple loop-mediated isothermal amplification (LAMP) chambers. Three bacterial strains (Escherichia coli O157:H7, methicillin-resistant Staphylococcus aureus and methicillin-sensitive S. aureus) were used to test the feasibility of the device. LAMP products were examined directly using a UV light and verified by agarose gel electrophoresis. Using this chip, we successfully detected E. coli O157:H7, MSSA and MRSA in less than 2 h. The detection limit for genes rfbE, spa and mecA (specific to E. coli O157:H7, MSSA and MRSA, respectively) was <10(2) CFU/100 μl. Further work is required to refine this approach and rigorously assess its analytical and diagnostic specificity and sensitivity.
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Yu
authors.affiliation: Beijing Institute of Respiratory Medicine, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Jiarui
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China.
authors.forename: Hualong
authors.initials: H
authors.lastname: Hao
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China. Electronic address: wenjiabin2005@sina.com.
authors.forename: Jiabin
authors.initials: J
authors.lastname: Wen
authors.affiliation: Department of Respiratory Medicine, The Second Hospital, Dalian Medical University, Dalian 116023, China. Electronic address: wqdlmu@163.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis*; D057230:Limit of Detection; D044085:Microfluidics / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods
chemical_list: D004269:DNA, Bacterial
keywords: Bacterial detection; Loop-mediated isothermal amplification (LAMP); Microfluidic chip; Solid phase extraction (SPE)