[Digoxigenin-labelled probes for detection of TEM-type beta-lactamases using the PCR technique].

BACKGROUND

Production of DNA probes is time-consuming and inefficient. We have developed a method for the obtention of digoxigenin-labeled probes to detect TEM-type betalactamases using the polymerase chain reaction. The amplification product was a 516 bp fragment internal to bla-TEM-1 from pBR 322.

METHODS

The techniques developed included extraction of plasmid DNA by lisis by alkali, electroelution, electrophoresis in agarose gels, polymerase chain reaction and hydridization with a DNA probe digoxigenin labeled.

RESULTS

We obtained by polymerase chain reaction 1500 ng of probe using 1 ng of target DNA. Developing classic methods the amount of probe was 75 ng from 1 microgram of target DNA. The time to obtain the probe was 3 hours, instead of a week with other methods.

CONCLUSIONS

We conclude that polymerase chain reaction is a good alternative to classic methods to obtain digoxigenin-labeled DNA probes.