Orion Corporation

Orion-Yhtyma Oy

Molecular Tool, Inc.

Orion-Yahtyma Oy

Hans E Soderlund

Tuula M Ranki

Anne-Christine Syvanen

Nisse E Kalkkinen

Airi M Palva

Arja M Weckman

Detection of variable nucleotide(s) is based on primer extension and incorporation of detectable nucleoside triphosphates. By selecting the detection step primers from the region immediately adjacent to the variable nucleotide, this variation can be detected after incorporation of as few as one nucleoside triphosphate. Labelled nucleoside triphosphates matching the variable nucleotide are added and the incorporation of a label into the detection step primer is measured. The selection of the detection step primer is important to the method according to this invention and is dependent on the nucleotide sequence of interest. The detection step primers are preferably selected so as to span the region immediately toward the 3' end from the variable nucleotide to be detected. The detection step primers can also be complementary to a sequence beginning several nucleotides removed from the variable nucleotide. The only limitation concerning the position of the detection step primers is that the sequence between the 3' end of the detection step primer and the variable nucleotide to be detected must not contain a nucleotide residue of the same type as the one to be detected. The detection step primers can equally well be chosen to be complementary to either the coding or non-coding strands of the nucleotide sequence of interest.

Method for determining specific nucleotide variations by primer

The present invention provides a rapid and sensitive method for assaying nucleic acids by means of hybridization techniques, wherein the detector probes are modified primers being incorporated into copies of the target nucleic acid before the hybridization reaction and a reagent combination as well as a kit therefor. The invention also provides a method for assaying nucleic acids by means of hybridization techniques, wherein the capturing probes are modified primers being incorporated into copies of the target nucleic acids before the hybridization reaction.

Method for assays of nucleic acid, a reagent combination and a kit

Amplification of nucleic acids is performed by incubating in a polymerization vessel a reaction mixture which contains in a suitable buffer solution, one or several single-stranded target nucleic acids, suitable primers, deoxyribouncleoside triphosphates and a polymerase. After a sufficient time for polymerication to occur, the reaction mixture is transferred into another vessel for the denaturation of the nucleic acids into single stranded nucleic acids. After denaturation, the reaction mixture is transferred back into the original vessel. The amplification process is regulated to maintain a temperature advantageous for the action of the polymerization enzyme in the polymerization vessel and a temperature advantageous for denaaturation in the denaturation vessel. Furthermore, the apparatus includes a liquid transfer system for transferring the reaction liquid from one vessel to another, which comprises at least one liquid transfer tube per vessel pair which extends from within the polymerization vessel to within the denaturation vessel.

Apparatus, for amplification of nucleic acids

The invention is related to improved nucleic acid reagents comprising arrays of nucleic acid fragments and combinations of such fragments. The preparation of such fragments by recombinant DNA techniques and their use in sandwich hybridization methods is also described. By making different combinations of the nucleic acid fragments--some labeled and some affixed to solid carriers, it is possible to create kits for the identification of e.g. veneral diseases. The improved nucleic acid reagents comprise two series of at least two alternating nucleic acid fragments, which are homologous to sequences in the nucleic acid to be identified, one of the series being labeled and one affixed to a solid carrier. Nucleic acid fragments belonging to different series must not be homologous to each other. Sandwich hybridization tests performed with arrays of nucleic acid fragments are at least four times as sensitive as sandwich hybridization tests performed with reagents belonging to the prior art.

Arrays of alternating nucleic acid fragments for hybridization arrays

This invention relates to a kit for the detection of microbial nucleic acids and a method for identifying the nucleic acids using a one-step sandwich hybridization technique. The technique requires two complementary nucleic acid reagents for each microbe or group of microbes to be identified.

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Espoo, Finland

Hans E Soderlund