Development of an enzyme linked immunosorbent assay to determine the numbers of chemolithotrophic bacteria at acid mine drainage sites.
USBM 1990 Jan; :1-42
Thiobacillus ferrooxidans is a prominent member of a group of chemolithotrophic bacteria that bear principal responsibility for the formation of acid mine drainage. A prototype enzyme-linked immunosorbent assay (ELISA) for emumerating and qualifying T. Ferrooxidans was assembled and characterized. The immunoassay protocol consisted of sequential incubations of the sample with (1) the primary antibody, (2) the enzyme-labeled secondary antibody, and (3) a chromogenic substrate specific for the enzyme label. The necessary reagents comprised primary polyclonal rabbit antibodies directed against T. Ferrooxidans ATCC 23270, alkaline phosphatase-coupled goat antirabbit polyclonal antibodies, and phenolphrhalein monophosphate. The elisa developed herein correctly identified whether iron-oxidizing bacteria were present in each of four samples supplied and analyzed by an independent laboratory. Sufficient preliminary data were obtained to warrant further research and development activities.
Bacteria; Microorganisms; Mining-industry; Waste-treatment; Analytical-processes; Analytical-methods
CP; Final Contract Report
NTIS Accession No.
U.S. Bureau of Mines
Oak Ridge Research Institute