title: [Rapid Detection of Adenovirus in Fecal Samples by Capillary Electrophoresis-laser Induced Fluorescence and Microchip Capillary Electrophoresis-laser Induced Fluorescence].
abstract: OBJECTIVE
  To establish a rapid and sensitive method based on polymerase chain reaction (PCR) combined with capillary electrophoresis-laser induced fluorescence (CE-LIF) and microchip capillary electrophoresis-laser induced fluorescence (MCE-LIF) for detecting adenoviruses in fecal samples.
  
  
  METHODS
  The DNA of adenovirus in fecal samples were extracted by the commercial kits and the conserved region of hexon gene was selected as the target gene and amplified by PCR reaction. After labeling highly sensitive nucleic acid fluorescent dye SYBR Gold and SYBR Orange respectively, PCR amplification products were separated by CE and MCE under the optimized condition and detected by LIF detector.
  
  
  RESULTS
  PCR amplification products could be detected within 9 min by CE-LIF and 6 min by MCE-LIF under the optimized separation condition. The sequenced PCR product showed good specificity in comparison with the prototype sequences from NCBI. The intraday and inter-day relative standard deviation (RSD) of the size (bp) of the target DNA was in the range of 1.14%-1.34% and 1.27%- 2.76%, respectively, for CE-LIF, and 1.18%-1.48% and 2.85%-4.06%, respectively, for MCE-LIF. The detection limits was 2.33 x 10(2) copies/mL for CE-LIF and 2.33 x 10(3) copies/mL for MCE-LIF. The two proposed methods were applied to detect fecal samples, both showing high accuracy.
  
  
  CONCLUSIONS
  The two proposed methods of PCR-CE-LIF and PCR-MCE-LIF can detect adenovirus in fecal samples rapidly, sensitively and specifically.
authors.forename: Jia
authors.initials: J
authors.lastname: Ruan
authors.forename: Dong-xia
authors.initials: DX
authors.lastname: Ren
authors.forename: Dan-ni
authors.initials: DN
authors.lastname: Yang
authors.forename: Pin-pin
authors.initials: PP
authors.lastname: Long
authors.forename: Hong-yue
authors.initials: HY
authors.lastname: Zhao
authors.forename: Yi-qi
authors.initials: YQ
authors.lastname: Wang
authors.forename: Yong-xin
authors.initials: YX
authors.lastname: Li
mesh_terms: D000256:Adenoviridae / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification; D019075:Electrophoresis, Capillary; D005243:Feces / Q000821:virology*; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral; D005456:Fluorescent Dyes