title: An integrated microfluidic system for diagnosis of the resistance of Helicobacter pylori to quinolone-based antibiotics.
abstract: Helicobacter pylori (H. pylori) is a species of bacteria that can colonize the human stomach mucosa. It is closely associated with gastric diseases such as ulcer and inflammation. Recently, some H. pylori strains were found to express resistance to a family of antibiotics known as quinolones due to single-point mutations. Although traditional polymerase chain reaction (PCR) and molecular diagnostic-based approaches can be used to determine the presence and abundance of antibiotic-resistant H. pylori strains, such processes are relatively expensive, labor-intensive, and require bulky and costly equipment. This study therefore reports an advanced diagnostic assay performed on an integrated microfluidic system for rapid detection of antibiotic resistance in H. pylori. The assay features three components: (1) nucleic acid extraction by specific probe-conjugated magnetic beads, (2) amplification of the target deoxyribonucleic acid (DNA) fragments by using single-nucleotide-polymorphism polymerase chain reaction (SNP-PCR), and (3) optical detection of the PCR products. The device integrates several microfluidic components including micro-pumps, normally-closed micro-valves, and reaction chambers such that the entire diagnostic assay can be automatically executed on a single microfluidic system within one hour with detection limits of 10(0), 10(2), and 10(2) bacterial cells for H. pylori detection and two different SNP sites strains. Three PCR-based assays for determining presence of H. pylori infection and two DNA single-point mutation assays aimed at determining whether the infected strains were resistant to quinolone can be performed simultaneously on a single chip, suggesting that this microfluidic system could be a promising tool for rapid diagnosis of the presence of antibiotic-resistant H. pylori strains.
authors.affiliation: Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chih-Yu
authors.initials: CY
authors.lastname: Chao
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chih-Hung
authors.initials: CH
authors.lastname: Wang
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Yu-Jui
authors.initials: YJ
authors.lastname: Che
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Cheng-Yen
authors.initials: CY
authors.lastname: Kao
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan; Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Jiunn-Jong
authors.initials: JJ
authors.lastname: Wu
authors.affiliation: Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan; Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan. Electronic address: gwobin@pme.nthu.edu.tw.
authors.forename: Gwo-Bin
authors.initials: GB
authors.lastname: Lee
mesh_terms: D015374:Biosensing Techniques; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D004352:Drug Resistance, Microbial / Q000235:genetics*; D016481:Helicobacter Infections / Q000188:drug therapy / Q000382:microbiology*; D016480:Helicobacter pylori / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D044085:Microfluidics / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D015363:Quinolones / Q000627:therapeutic use
chemical_list: D004269:DNA, Bacterial; D015363:Quinolones
keywords: Antibiotic resistance detection; H. pylori; Microfluidic; Optical detection; SNP-PCR