title: Microscale sample preparation for PCR of C. difficile infected stool.
abstract: In this paper, we describe the design of a microfluidic sample preparation chip for human stool samples infected with Clostridium difficile. We established a polymerase chain reaction able to distinguish C. difficile in the presence of several other organisms found in the normal intestinal flora. A protocol for on-chip extraction of nucleic acids from clinical samples is described that can detect target DNA down to 5.0x10(-3) ng of template. The assay and sample preparation chip were then validated using known positive and known negative clinical samples. The work presented has potential applications in both the developed and developing world.
authors.affiliation: Department of Medicine, Section of Gastroenterology, Boston University School of Medicine, Suite 504, 650 Albany Street, Boston, MA 02118, United States.
authors.forename: Sara
authors.initials: S
authors.lastname: Gillers
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Atkinson
authors.forename: Aaron C
authors.initials: AC
authors.lastname: Bartoo
authors.forename: Madhumita
authors.initials: M
authors.lastname: Mahalanabis
authors.forename: Michael O
authors.initials: MO
authors.lastname: Boylan
authors.forename: John H
authors.initials: JH
authors.lastname: Schwartz
authors.forename: Catherine
authors.initials: C
authors.lastname: Klapperich
authors.forename: Satish K
authors.initials: SK
authors.lastname: Singh
mesh_terms: D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial