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19-Feb-2014
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Arch Hellen Med, 31(1), January-February 2014, 23-40 REVIEW Molecular techniques for the detection and characterization of microorganisms I.D. Kyriazis,1,2 M.E. Kambouris,1 K. Poulas,1 G.P. Patrinos1 |
The detection and characterization of pathogenic and non-pathogenic microorganisms are currently of paramount importance in clinical and laboratory practice. Molecular techniques can offer more accurate characterization, increase sensitivity and reduce the diagnosis time without substantial increase in cost. They thus result in faster and more appropriate treatment and also promote prevention procedures, guiding decisions on the introduction and composition of prophylactic therapy. The discovery of the polymerase chain reaction (PCR) and other similar molecular techniques, based on amplification of nucleic acid traces, has contributed to the rapid growth of diagnostic methodology, as real-time PCR enables the simultaneous qualitative and quantitative determination of microorganisms. Novel and modified molecular techniques are being evolved to improve reliability and enhance sensitivity. These are based on new insights into the chemistry of synthesis and detection and the use of improved incubation and detection hardware. The majority of the new techniques are only gradually being introduced into the routine of diagnostic laboratories, because of the domination of the PCR, the need for expensive specialized laboratory equipment and instrumentation, which in turn require experienced, expert personnel for effective operation, and their high overall operating cost of, taking into consideration the reagents and the maintenance costs. The need for standardization, if not of techniques, at least operating procedures and the recording of data to enable comparison, cooperation and interaction among centers further complicates the choice and applicability of these new and promising, but also unproven methods. This review presents the main molecular diagnostic methods used in microbiology laboratories.
Key words: Microbiology, Molecular diagnosis.