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Development of an Enzyme Linked Immunosorbent Assay to Determine the Numbers of Chemolithotrophic Bacteria at Acid Mine Drainage Sites.


PB91146126

Publication Date 1990
Personal Author Blake, R. C.; Revis, N. W.; Holdsworth, G.
Page Count 50
Abstract Thiobacillus ferrooxidans is a prominent member of a group of chemo-lithotrophic bacteria that bear principal responsibility for the formation of acid mine drainage. A prototype enzyme-linked immunosorbent assay (ELISA) for enumerating and qualifying T. ferrooxidans was assembled and characterized. The immunoassay protocol consisted of sequential incubations of the sample with (i) the primary antibody, (ii) the enzyme-labeled secondary antibody, and (iii) a chromogenic substrate specific for the enzyme lable. The necessary reagents comprised primary polyclonal rabbit antibodies directed against T. ferrooxidans ATCC 23270, alkaline phosphatase-copled goat anti-rabbit polyclonal antibodies, and phenolphrhalein monophosphate. The ELISA developed herein correctly identified whether iron-oxidizing bacteria were present in each of 4 samples supplied and analyzed by an independent laboratory. Sufficient preliminary data was obtained to warrant further research and development activities.
Keywords
  • Chemical water pollution
  • Mineral acids
  • Coal mining
  • Enzyme-linked immunosorbent assay
  • Mine waters
  • Chemical analysis
  • Substrate specificity
  • Thiobacillus ferrooxidans
  • Mine drainage sites
  • Polyclonal antibodies
Source Agency
  • Bureau of Mines
NTIS Subject Category
  • 68D - Water Pollution & Control
  • 48A - Mineral Industries
  • 57F - Cytology, Genetics, & Molecular Biology
Corporate Authors Oak Ridge Research Inst., TN.; Bureau of Mines, Pittsburgh, PA. Pittsburgh Research Center.
Supplemental Notes Sponsored by Bureau of Mines, Pittsburgh, PA. Pittsburgh Research Center.
Document Type Technical Report
Title Note Technical rept. (Final).
NTIS Issue Number 199110
Contract Number
  • J0388014
Development of an Enzyme Linked Immunosorbent Assay to Determine the Numbers of Chemolithotrophic Bacteria at Acid Mine Drainage Sites.
Development of an Enzyme Linked Immunosorbent Assay to Determine the Numbers of Chemolithotrophic Bacteria at Acid Mine Drainage Sites.
PB91146126

  • Chemical water pollution
  • Mineral acids
  • Coal mining
  • Enzyme-linked immunosorbent assay
  • Mine waters
  • Chemical analysis
  • Substrate specificity
  • Thiobacillus ferrooxidans
  • Mine drainage sites
  • Polyclonal antibodies
  • Bureau of Mines
  • 68D - Water Pollution & Control
  • 48A - Mineral Industries
  • 57F - Cytology, Genetics, & Molecular Biology
  • J0388014
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