Title : Development of LAMP Kit for Rapid pathogen Detection in Neonatal Sepsis
Abstract:
Delayed detection and treatment of sepsis in newborns (neonatal sepsis) is one of the leading causes of high infant mortality rates in India. We are developing a cost-effective LAMP based amplification kit that can be used for early diagnosis of bacterial infection that causes sepsis in newborns. The detection can be done from the patient blood/urine samples. This LAMP based kit does not require skilled expertise or a complex lab infrastructure and can be used in low resource settings (small clinics and in field). The assay will take 45 to 60 minutes for fast and accurate diagnosis. Blood culture is the only routine method available for identification of bacteria. However, the method is time consuming and takes 4 to 5 days for the result. It requires sophisticated infrastructure with skilled expertise, which is not available in many hospitals across the country. Since, accurate and early diagnosis of bacteria directly from patient’s blood/urine has a great potential, we are developing a LAMP kit for rapid detection of multiple bacteria involved in neonatal sepsis. We have optimized our assay for specific detection of seven different bacteria such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Streptococcus agalactiae, Enterobacter cloacae, Klebseilla pneumonia, Staphylococcus epidermidis or assay showed a great specificity with a consistent detection limit ~104 CFU/mL-1 for each organism. Also the assay exhibited a sensitivity limit ranging from 50 to 100 picograms for the detection of bacteria from DNA. The objective is to develop the kit as a point of-care diagnostic kit, which will help clinicians in selecting right antibiotic therapy for timely treatment.
