Abstract: The present invention relates to a process for conducting real-time PCR, and to a device for conducting the method of the present invention. The invention is especially suited for the simultaneous identification and quantification of nucleic acids present in a sample, e.g. a biological sample. Further, this invention describes a method for simultaneous quantitative analysis of multiple nucleic acid sequences in a single compartment by using an integrated nucleic acid microarray combined with a highly surface-specific readout device. The invention relates to a device wherein a surface which is either part of the chamber surface or a surface that is created in the reaction chamber, such as bead surface, is coated with capture probes and in the same chamber, a PCR reaction takes place.

Abstract: The invention provides a method for quantifying an initial ratio of the amounts of at least two nucleic acids of interest in a sample by means of a multiplex nucleic acid amplification reaction, comprising amplifying the nucleic acids of interest in the amplification reaction, measuring the amount of at least two nucleic acids of interest at at least two different time points in the reaction, determining from at least two of the measurements the amplification rate of the at least two nucleic acids of interest, comparing the rates with a reference, and determining from the comparison the initial ratio of the amounts of the at least two nucleic acids of interest in the sample. Preferably, at least one variable factor in the nucleic acid amplification reaction is adjusted in order to allow detectable levels of all nucleic acids of interest to be reached before an amplification and/or detection limit of one or more of the nucleic acids of interest is reached.

Abstract: Statins are widely used for the treatment of high cholesterol levels. In the present invention, the effect of statins on mitochondrial nucleic acid and the activation state of mitochondria is used in methods for determining whether a subject is at risk of developing side effects of statin treatment, methods for the treatment of a clinical symptom associated with reduced mitochondrial function. Further provided are kits and the like comprising a means for the detection of mitochondrial nucleic acid, or the activation of a mitochondrion for use in a method mentioned above.

Abstract: The invention relates to the diagnosis of disease or the determination of functioning of cellular organisms, being of multi-cellular or unicellular nature, being visible by the naked eye or being a microorganism. The invention provides a method for determining functioning of a cellular organism comprising determining the relative ratio of a first endosymbiont cellular organelle nucleic acid and/or gene product thereof in a sample obtained from the organism in relation to the amount of a second nucleic acid and/or gene product thereof.

Abstract: The invention relates to the diagnosis of disease or the determination of functioning of cellular organisms, being of multi-cellular or unicellular nature, being visible by the naked eye or being a microorganism. The invention provides a method for determining functioning of a cellular organism comprising determining the relative ratio of a first endosymbiont cellular organelle nucleic acid and/or gene product thereof in a sample obtained from the organism in relation to the amount of a second nucleic acid and/or gene product thereof.

Abstract: Methods for amplifying nucleic acid in a sample including providing the sample with a set of primers to enable synthesis of at least one nucleic acid strand complementary to at least part of the nucleic acid, wherein the set of primers comprises between 3-8 random bases, preferably clustered near the 3? end of each primer in said set of primers. The methods of the invention are useful, for example, for determining whether samples derived from humans, mammals, poultry, or fish comprise nucleic acid of a pathogen. The methods are further suited for typing the pathogen and typing particular variants of said pathogen. The methods are also suited for the elucidation of the gene expression profile or genetic profile of cells.

Abstract: The invention provides a method for reducing background in hybridization reactions of nucleic acids involving at least two homologous probes, wherein at least one of the probes is nonlinear, or two homologous target sequences and a nonlinear probe. Background is reduced by introducing an intended mismatch with a target sequence in at least one of the probes. The presence of the mismatch reduces the specificity of probes not entirely complementary to a target sequence to such an extent that the background signal is reduced. A set of mixed homologous probes, wherein at least one of the probes is nonlinear, comprising such specific mismatch is also provided. The set can be used for the detection of variants of a family of nucleic acids, for instance, a number of HIV variants. The invention also provides kits for carrying out the methods according to the invention.

Abstract: Methods for amplifying nucleic acid in a sample comprising providing the sample with a set of primers to enable synthesis of at least one nucleic acid strand complementary to at least part of the nucleic acid, wherein the set of primers comprises between 3-8 random bases, preferably clustered near the 3′ end of each primer in said set of primers. The methods of the invention are useful, for example, for determining whether samples derived from humans, mammals, poultry, or fish comprise nucleic acid of a pathogen. The methods are further suited for typing the pathogen and typing particular variants of said pathogen. The methods are also suited for the elucidation of the gene expression profile or genetic profile of cells.

Abstract: The invention provides a method for quantifying an initial ratio of the amounts of at least two nucleic acids of interest in a sample by means of a multiplex nucleic acid amplification reaction, comprising amplifying the nucleic acids of interest in the amplification reaction, measuring the amount of at least two nucleic acids of interest at at least two different time points in the reaction, determining from at least two of the measurements the amplification rate of the at least two nucleic acids of interest, comparing the rates with a reference, and determining from the comparison the initial ratio of the amounts of the at least two nucleic acids of interest in the sample. Preferably, at least one variable factor in the nucleic acid amplification reaction is adjusted in order to allow detectable levels of all nucleic acids of interest to be reached before an amplification and/or detection limit of one or more of the nucleic acids of interest is reached.

Abstract: The invention provides a method for reducing background in hybridization reactions of nucleic acids involving at least two homologous probes, wherein at least one of the probes is non-linear, or two homologous target sequences and a non-linear probe. Background is reduced by introducing an intended mismatch with a target sequence in at least one of the probes. The presence of the mismatch reduces the specificity of probes not entirely complementary to a target sequence to such an extent that the background signal is reduced. A set of mixed homologous probes, wherein at least one of the probes is non-linear, comprising such specific mismatch is also provided. The set can be used for the detection of variants of a family of nucleic acids, for instance a number of HIV variants. The invention also provides kits for carrying out the methods according to the invention.

Abstract: The present invention is concerned with a method for generating, in a non specific manner, multiple copies of RNA from a pool of mRNA's. Such a method is of particular importance in techniques for screening the differences in expression in given cell types or in cells under specific conditions. The present invention provides a non-selective poly A mRNA labeling and amplification method, i.e. a method not encompassing cDNA synthesis.

Abstract: Methods for amplifying nucleic acid in a sample comprising providing the sample with a set of primers to enable synthesis of at least one nucleic acid strand complementary to at least part of the nucleic acid, wherein the set of primers comprises between 3-8 random bases, preferably clustered near the 3′ end of each primer in said set of primers. The methods of the invention are useful, for example, for determining whether samples derived from humans, mammals, poultry, or fish comprise nucleic acid of a pathogen. The methods are further suited for typing the pathogen and typing particular variants of said pathogen. The methods are also suited for the elucidation of the gene expression profile or genetic profile of cells.

Abstract: The present application relates to mutated RNA polymerases from bacteriophages that have increased stability, for example under high temperature conditions. One example of bacteriophage encoded RNA polymerase is the T7 RNA polymerase. T7 is a bacteriophage capable of infecting E. coli cells. Examples of other E. coli infecting T7-like bacteriophages are T3, øI, øII, W31, H, Y, A1, croC21, C22 and C23. An example of a Salmonella typhimurium infecting bacteriophage is SP6. The present invention is concerned with the RNA polymerases of T7-like bacteriophages that have been mutated. Due to these mutations the RNAP's have an increased stability. Preferred mutated RNA polymerases according to the invention are mutant RNA polymerases from T7 or SP3 bacteriophages. Due to the high homology between these enzymes, mutations in the T7 gene 1 sequence are likely to have the same effect in the corresponding gene sequence of the T3 bacteriophage.

Abstract: The invention relates to the diagnosis of disease or the determination of functioning of cellular organisms, being of multi-cellular or unicellular nature, being visible by the naked eye or being a microorganism. The invention provides a method for determining functioning of a cellular organism comprising determining the relative ratio of a first endosymbiont cellular organelle nucleic acid and/or gene product thereof in a sample obtained from said organism in relation to the amount of a second nucleic acid and/or gene product thereof.

Abstract: The present invention relates to a method for isolating nucleic acid from a sample. Known methods for the isolation of nucleic acid from these materials may comprise the use of a chaotropic substance, for example to lyse cells present in the starting material, and free the nucleic acid therefrom. When a nucleic acid containing material is treated with a chaotrope and contacted with a nucleic acid binding solid phase, the nucleic acid will bind to the solid phase in the presence of the chaotrope and the solid phase with the nucleic acid bound thereto can thus be separated from the remainder of the sample.

Abstract: The present invention is concerned with a method for generating, in a non specific manner, multiple copies of RNA from a pool of mRNA's. Such a method is of particular importance in techniques for screening the differences in expression in given cell types or in cells under specific conditions. The present invention provides a non-selective poly A mRNA labeling and amplification method, i.e. a method not encompassing cDNA synthesis.

Abstract: The present invention is concerned with a method for generating, in a non specific manner, multiple copies of RNA from a pool mRNA's. Such a method is of particular importance in techniques for screening the differences in expression in given cell types or in cells under specific conditions. The present invention provides a non-selective poly A mRNA labeling and amplification method, i.e. a method not encompassing cDNA synthesis.

Abstract: The invention provides a method for reducing background in hybridization reactions of nucleic acids involving at least two homologous probes, wherein at least one of the probes is non-linear, or two homologous target sequences and a non-linear probe. Background is reduced by introducing an intended mismatch with a target sequence in at least one of the probes. The presence of the mismatch reduces the specificity of probes not entirely complementary to a target sequence to such an extent that the background signal is reduced. A set of mixed homologous probes, wherein at least one of the probes is non-linear, comprising such specific mismatch is also provided. The set can be used for the detection of variants of a family of nucleic acids, for instance a number of HIV variants. The invention also provides kits for carrying out the methods according to the invention.

Abstract: The present invention is directed to a transcription based amplification method for the amplification of DNA targets. With the method of the present invention an isothermal transcription based amplification method is provided for the amplification of double stranded DNA.

Abstract: The present invention provides oligonucleotides that can be used as primers to amplify a region of the 16S rRNA of M. pneumoniae. Also provided are probes and kits for detection of amplified RNA and typing of M. pneumoniae strains. The primers, probes, methods and kits are useful for diagnosing M. pneumoniae.