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A Membrane-Array Method to Detect Specific Human Intestinal Bacteria in Fecal Samples Using Reverse Transcriptase-PCR and Chemiluminescence
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  • A Membrane-Array Method to Detect Specific Human Intestinal Bacteria in Fecal Samples Using Reverse Transcriptase-PCR and Chemiluminescence
  • A Membrane-Array Method to Detect Specific Human Intestinal Bacteria in Fecal Samples Using Reverse Transcriptase-PCR and Chemiluminescence
저자명
KIM. PYOUNG IL,ERICKSON. BRUCE D,CERNIGLIA. CARL E.
간행물명
Journal of microbiology and biotechnology
권/호정보
2005년|15권 2호|pp.310-320 (11 pages)
발행정보
한국미생물생명공학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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A membrane-based oligonucleotide array was used to detect predominant bacterial species in human fecal samples. Digoxygenin-labeled 16S rDNA probes were generated by PCR from DNA that had been extracted from fecal samples or slurries. These probes were hybridized to an array of 120 oligonucleotides with sequences specific for 40 different bacterial species commonly found in human feces, followed by color development using an alkaline phosphatase-conjugated antibody and NBT /BCIP. Twenty of the species were detected by this method, but E. coli, which was present at $~$1 $ imes 10$^5$ CFU per gram feces, was not detected. To improve the sensitivity of this assay, reverse transcriptase-PCR was used to generate probes from RNA extracted from fecal cultures. Coupled with a chemiluminescence detection method, this approach lowered the detection limit for E. coli from $~1$ $ imes 10$^6$ to ${leq}$ 1 $ imes 10$^5$ These results indicate that the membrane-array method with reverse transcriptase-PCR and chemiluminescence detection can simultaneously identify bacterial species present in fecal samples at cell concentrations as low as${leq}$ 1 $ imes 10$^5$ CFU per gram.