BACTERIAL VIABILITY IN EARLY NEONATAL SEPSIS EVALUATED BY RT-PCR

Abstract Introduction: Neonatal sepsis remains a serious and potentially fatal disease. One of the most challenging aspects in the management of sepsis neonates is their diagnosis. Blood cultures may fail to reveal the microorganism in symptomatic patients. The importance of an accurate and rapid method with effective sensitivity to detect and prevent the indiscriminate use of antibiotics appears. The final approach is to reduce morbidity and mortality of newborns suspected of early neonatal sepsis. Objective: Evaluation of cDNAs of the main microorganisms involved in PNS from total RNA of E. coli , S. aureus and S. agalactiae , using reverse transcription followed by real-time PCR. Methods: Specific oligonucleotide primers were designed and synthesized to amplify target amplicons of genes selected from isolated microorganisms. Total RNA was extracted from each microorganism for RT-qPCR. Calibration curves and cDNA detection of viable bacteria were determined. Results: obtaining bacterial cDNAs allowed the construction of standard curves by RT-qPCR of collection samples; The data obtained by the cDNA dilution curves show that the use for blood detection and identification of bacteria in the tested samples is pertinent. Conclusion: The results of the present study present an RT-specific molecular tool of bacterial RNA followed by qPCR, potentially efficient and important for detecting bacteremia in which the microorganism is alive. This method should be considered in the future as an important diagnostic tool for small amounts of blood in patients with early neonatal sepsis. The RT method can help prevent blind antibiotic use.