1887

Abstract

SUMMARY

A dot-blot hybridisation assay for the detection of was used to compare a P-labelled probe with two biotinylated probes and a sulphonated probe. The minimum amount of pure DNA that could be detected by the P-labelled probe was 100 pg, which corresponded to 5 × 10 bacteria. A biotin-labelled DNA (biotin-DNA) probe together with the BluGene detection system produced by Bethesda Research Laboratories (BRL), and a sulphonated probe and Chemiprobe detection system (Orgenics) gave similar levels of sensitivity; nylon membranes could be used with both these non-radioactive detection systems. However, a photobiotin-labelled DNA (photobiotin-DNA) probe and detection system produced by Biotechnology Research Enterprises S.A. (BRESA) gave optimum results only with nitrocellulose membranes, and was quantitatively 100 times less sensitive than the other types of probe. The detection systems for the biotin-DNA and photobiotin-DNA probes produced non-specific reactions with crude bacterial blots of heterologous organisms; these non-specific reactions could be removed by treating the dot blots with proteinase K, but not by treatment with RNAase. The sulphonated probe and detection system did not give any reaction with heterologous organism blots.

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1988-08-01
2022-01-17
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