Rapid Assays for the Detection and Determination of Sparse Populations of Bacteria and Bacteriophage T7 with Radioactively Labelled Homologous Antibodies
SUMMARY: Small numbers of several bacterial species were rapidly and specifically detected with the 125I-labelled antibody method described previously. Multibacterial species detection was achieved in one operation with 125I-labelled mixtures of type-specific anti-bacterial globulins but the level and variability of the blank value increased, and sensitivity decreased, with the number of globulin components in the mixture. An indirect radio-assay that involved successive treatments of bacteria with unlabelled rabbit anti-bacterial serum and 125I-labelled immunopurified goat anti-rabbit globulin was less sensitive and reproducible than the direct radio-assay. A modified assay, developed to detect and determine bacteria filtered on to a membrane filter, allowed the detection of small numbers of bacteria in large volumes of aqueous sample.
The feasibility of rapidly and specifically detecting small numbers of virus particles with 125I-labelled anti-viral globulin was investigated with bacteriophage T7 as a model particle. Methods for the rapid separation of phage-125I-labelled globulin complex were developed and a minimum of about 5–105 total phage particles was detected.
AVRAMEASS.,
TERNYNCKT.1969; The cross-linking of proteins with glutaraldehyde and its use for the preparation of immunoadsorbents.. Immunochemistry 6:53–66
CLOSSO.1968; Rapid identification of small numbers of bacteria after short time cultivation in the presence of FITC-conjugated antiserum.. Acta pathologica microbiologica scandinavica 72:412–420
COOMBSR. R. A.,
MOURANTA. E.,
RACER. R.1945; A new test for the detection of weak and ‘incomplete’ Rh agglutinins.. British Journal of Experimental Pathology 26:255–266
DANIELSSOND.1965; A membrane filter method for the demonstration of bacteria by the fluorescent antibody technique. I.A methodological study.. Acta pathologica microbiologica scandinavica 63:597–603
DANIELSSOND.,
LAURELLG.1965; A membrane filter method for the demonstration of bacteria by the fluorescent antibody technique. 2. The application of the method for detection of small numbers of bacteria in water.. Acta pathologica microbiologica scandinavica 63:604–608
GUTHRIER. K.,
REEDERD. J.1969; Membrane filter-fluorescent-antibody method for detection and enumeration of bacteria in water.. Applied Microbiology 17:399–401
JOSTR.,
FEYH.1971; Rapid detection of small numbers of airborne bacteria by a membrane filter fluorescent-antibody technique.. Applied Microbiology 20:861–863
LOWRYO. H.,
ROSEBROUGHN. J.,
FARRA. L.,
RANDALLR. J.1951; Protein measurement with the Folin. phenol reagent.. Journal of Biological Chemistry 193:265–275
STRANGER. E.,
POWELLE. O.,
PEARCET. W.1971; The rapid detection and determination of sparse bacterial populations with radioactively-labelled homologous antibodies.. Journal of General Microbiology 67:349–357
Rapid Assays for the Detection and Determination of Sparse Populations of Bacteria and Bacteriophage T7 with Radioactively Labelled Homologous Antibodies