Biological And Molecular Test Methods For Detection Of Sags And Sag Carrying S Pyogenes

The detection and assay of SAgs in bacterial supernatants, purified preparations, or biological fluids (e.g., patient plasma) are based on the classical mitogen-induced lymphocyte proliferation test. Human peripheral blood lymphocytes, purified (Ficoll gradient) from the blood of healthy donors, are incubated in multiwell microtiter plates at 10"5 cells per well. After 48 hr, 0.1 mCi of tritiated thymidine is added for another 24 hr. The cells are harvested and thymidine incorporation is measured in a radioactivity scintillation counter.[4] This procedure is also used by testing the inhibition of SAg-induced lymphocyte proliferation by specific antibodies against the relevant SAg.[4]

The determination of SAg-specific TCR Vp pattern is investigated by Vp enrichment analysis using anchored multiprimer amplification.1-15-1

The identification of SAg genes in S. pyogenes isolates is undertaken by polymerase chain reaction (PCR). Bacterial isolates are grown overnight in brain-heart infusion medium. The bacterial cells are spun down and washed. Their DNA is purified and used for PCR with specific primers for the various spe genes. A primer pair specific to a DNA region encoding the 23S rRNA is used as a positive control.[4]

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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