Abstract :This work has as its main objective the development of a more comprehensive molecular diagnostic
method, efficient to diagnose crucial agents in early neonatal bacterial sepsis (ENS), in order to
elaborate a rational and specific therapy. Genomic material extracted from the target bacteria
Streptococcus agalactiae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa,
Enterobacter sp, Serratia sp and Staphylococcus aureus, were analyzed by multiplex real-time
Polymerase Chain Reaction (qPCR) from samples of DNA from cultures of microorganisms
obtained in collection stocks, leukocyte DNA samples from negative and positive control donor
individuals. In this study positivity was found by multiplex qPCR in all samples containing the
presence of bacterial DNA. The development of this test, based on multiplex qPCR, for the rapid
detection (results obtained up to 6 hours) and sensitive of important pathogens causing ENS could
potentially outperform in comparison to microbiological methods that are based on bacterial
culture. This approach was designed to facilitate progression to specific antimicrobial therapy, which
is especially relevant in term and preterm newborns, where empirical treatment is generally used
empirically in neonatal intensive care units. The test developed based on Multiplex qPCR proved to
be adequate and sensitive for the diagnosis of the presence of genomic DNA of the bacteria studied
and that the adaptation of this method has the potential to result in higher rates of positivity to
detect the bacteria studied.