Novel SYBR Green Real Time PCR Method for Direct Quantification and Detection of Escherichia coli from Clinical Samples

Abstract

Diagnosis of sepsis in its early stages are key steps for its treatment, however because of the brutal effects of sepsis and the necessity of its immediate treatment, many physicians don’t wait for the conventional microbiological diagnosis and start prescribing antimicrobial drugs. Therefore designing an accurate, fast, reliable and economic method to quantify the total number of Escherichia coli from blood and clinical samples by quantitative SYBR green real time PCR is a key step for early diagnosis and treatment of the problem and is compatible with alternative conventional microbiological methods. The method included artificial inoculation of healthy blood with type strain E. coli used for designing a quantitative standard curve. The protocol was then applied on blood samples collected from suspected patients with sepsis, also used for clinical and environmental samples. SYBR green used as inexpensive indicator to reduce the costs while using probe based methods.