Microscale sample preparation for PCR of C. difficile infected stool.
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.
Gillers
Sara
S
Department of Medicine, Section of Gastroenterology, Boston University School of Medicine, Suite 504, 650 Albany Street, Boston, MA 02118, United States.
Atkinson
Christopher D
CD
Bartoo
Aaron C
AC
Mahalanabis
Madhumita
M
Boylan
Michael O
MO
Schwartz
John H
JH
Klapperich
Catherine
C
Singh
Satish K
SK
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
D004269:DNA, Bacterial