Am J Clin Microbiol Antimicrob | Volume 1, Issue 1 | Research Article | Open Access
Walter Peres da Silva*, Durval Batista Palhares, Paola Mendes Burkhardt and Almir de Sousa Martins
Laboratory for Research in Pediatrics, University Hospital of UFMS, Campo Grande, Brasil
*Correspondance to: Walter Peres da Silva
Fulltext PDFThis 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.
Multiplex qPCR; Molecular diagnosis; Bacterial pathogens
da Silva WP, Palhares DB, Burkhardt PM, de Sousa Martins A. Heptaplex Real-Time Polimerase Chain Reaction for Neonate Sepsis Pathogens in Brazil. Am J Clin Microbiol Antimicrob. 2018; 1(1): 1003.