Cs. Bernal et al., DEVELOPMENT OF A SENSITIVE AND QUANTITATIVE ENZYME-LINKED IMMUNOFILTER ASSAY (ELIFA) FOR WHOLE BACTERIAL-CELLS, Journal of microbiological methods, 19(2), 1994, pp. 135-143
The application of enzyme-linked immunofilter assay (ELIFA) techniques
to the detection of whole bacterial cells in aqueous samples is descr
ibed. In a qualitative assay, Escherichia coli cells were collected on
25 mm diameter cellulose ester membranes in filter holders and treate
d in situ with rabbit anti-E. coli IgG, phosphatase-conjugated anti-ra
bbit IgG and chromogenic enzyme substrate. The presence of 10(4) bacte
ria per membrane was detected by colour development (assessed visually
) in a total time of 2 h 15 min. In,a modification of the procedure, m
embrane-bottomed plates containing 96 x 6 mm diameter wells were used
and after application of the enzyme substrate the membrane from each p
late was peeled off and scanned at 626 nm. This procedure enabled the
quantitative detection of 10(3) to 10(7) bacteria per membrane in a ti
me of 1 h 45 min.