MAGNETIC SEPARATION TECHNIQUES IN DIAGNOSTIC MICROBIOLOGY

The principles of magnetic separation aided by antibodies or other spe cific binding molecules have been used for isolation of specific viabl e whole organisms, antigens, or nucleic acids. Whereas growth on selec tive media may be helpful in isolation of a certain bacterial species, immunomagnetic separation (IMS) technology can isolate strains posses sing specific and characteristic surface antigens. Further separation, cultivation, and identification of the isolate can be performed by tr aditional biochemical, immunologic, or molecular methods. PCR can be u sed for amplification and identification of genes of diagnostic import ance for a target organism. The combination of IMS and PCR reduces the assay time to several hours while increasing both specificity and sen sitivity. Use of streptavidin-coated magnetic beads for separation of amplified DNA fragments, containing both biotin and a signal molecule, has allowed for the conversion of the traditional PCR into an easy-to -read microtiter plate format The bead-bound PCR amplicons can also ea sily be sequenced in an automated DNA sequencer. The latter technique makes it possible to obtain sequence data of 300 to 600 bases from 20 to 30 strains, starting with clinical samples, within 12 to 24 h. Sequ ence data can be used far both diagnostic and epidemiologic purposes. IMS has been demonstrated to be a useful method in diagnostic microbio logy. Most recent publications describe IMS as a method for enhancing the specificity and sensitivity of other detection systems, such as PC R, and providing considerable savings in time compared with traditiona l diagnostic systems. The relevance to clinical diagnosis has, however , not yet been fully established for all of these new test principles. In the case of PCR, for example, the presence of specific DNA in a fo od sample does not demonstrate the presence of a live organism capable of inducing a disease. However, all tests offering increased sensitiv ity and specificity of detection, combined with reduced time of analys is, have to be seriously evaluated.