BRUCELLA SPP SPECIFIC DNA SEQUENCES DETECTION PROCEDURE THROUGH POLYMERASE CHAIN (PCR) REACTION COUPLED TO AN ENZYMOINMUNOENSAY (ELISA).

摘要

Method for detecting specific DNA sequences from Brucella spp. by polymerase chain reaction (PCR) coupled to an enzyme immunoassay (ELISA). After selective amplification of a Brucella 223 bp sequence using oligonucleotides B4 and B5, the resulting amplified products labeled with digoxigenin (DIG) are hybridized with a biotinylated capture probe. Subsequently, these hybrids are captured on streptavidin coated plates and detected using an anti-DIG Fab / peroxidase conjugate. The proposed technique has been much more sensitive than conventional PCR and that bacteriological methods and more specific than usual serological methods, allowing its use for the implementation of an easy and rapid molecular diagnostic procedure for Brucella spp infection . in human blood samples and in other clinical samples, the monitoring of the response to treatment and the early detection of recurrences avoiding the risk of germ manipulation.