FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biochemistry, more specifically to methods of carrying out immunoenzymometric and DNA-hybridisation analysis. A conjugate is proposed for bioluminescent molecular microanalysis, obtained from an enzyme, which is bonded to a biospecific reagent, where the enzyme used is Ca2+ regulated photoprotein obelin with an altered bioluminescent spectrum: recombinant obelin W92F; H22E or recombinant obelin Y138F. The method for simultaneous detection of two analytes through bioluminescent molecular microanalysis involves treating a solid phase with biospecific reagents, separation of unreacted liquid phase, treatment of the solid phase with an enzyme containing conjugate, separation of the liquid and solid phases and analysis of the solid phase. The biospecific reagent used for the first treatment of the solid phase is two immumoglobulins or one immunoglobulin with double specificity or two different oligonucleotides. Final treatment of the solid phase is done simultaneously with two conjugates, which respectively consist of recombinant obelins with different bioluminescent spectra and molecules with different biospecificity, where the recombinant obelins used are W92F;H22E and recombinant obelin Y138F with subsequent analysis on a double-channel bioluminometre with broad-band light filters.
EFFECT: use of this invention allows for accurate, fast, universal and easy realisation of the method of bioluminescent molecular microanalysis.
4 ex, 3 dwg, 2 cl
