New ultra rapid test for detection of bacteria

• Published in: Pathologie biologie (Paris) 2011-10, Vol.59 (5), p.248-255
• Main Authors: Goldschmidt, P, Balloy, T, Degorge, S, Benallaoua, D, Batellier, L, Koskas, F, Borsali, E, Chaumeil, C
• Format: Article
• Language: fre
• Subjects: Bacteria - classification; Bacteria - genetics; Bacteria - isolation & purification; DNA, Bacterial - analysis; Humans; Real-Time Polymerase Chain Reaction - methods; Time Factors
• ISSN: 1768-3114
• DOI: 10.1016/j.patbio.2009.10.008

Bacteriological testing is aimed to reduce the risk of transmission of infections. However, the detection of Bacteria by culture requires from 18hours to 14 days and may produce erroneous results for fastidious species. The goal of this work was to design and validate a new tool for bacterial testing. The test is based on the fast real-time PCR (frt PCR). The DNA extracted from samples containing internal controls are introduced into four tubes containing primers and probes for the frt PCR. The cycling program consists in 1×at 95°C for 10min and 45×(15s at 95°C, 8s) at 52°C and 10s at 72°C. The frt PCR detects 0,01 CFU/μl of Bacteria and identifies eight Genera without interferences from the environment or from fungi and with no need for melting curve analysis or additional sequencing. The frt PCR detects and quantifies Bacteria identifying and assessing the load of Staphylococci, Streptococci, Haemophilus, Pseudomonas, Enterobacteria, Acinetobacter, Propionibacteriacae and Corynebacteria. Cultures require at least 24hours but the new frt PCR reduces the time to 90minutes. Larger series of samples are necessary to confirm the usefulness of this new test for routine bacterial sterility controls.