Abstract: Described is a method for quantifying nucleic acid in a nucleic acid amplification reaction, in which an asymmetric nucleic acid amplification reaction is performed on a sample such that double stranded nucleic acid product is generated in an initial stage of the reaction, and single stranded nucleic acid product is generated in a subsequent stage of the reaction once a limiting primer is exhausted. Relative amounts of double stranded nucleic acid product and single stranded nucleic acid product produced are then detected by means of a melt curve analysis. The ratio of double stranded product peak height to single stranded product peak height may then be used to quantify the amount of starting template based on the ratio of double stranded product peak height to single stranded product peak height.

Abstract: We describe a quantitative PCR (qPCR) instrument for combined qPCR and melt curve (dissociation and/or association curve) analysis. The instrument has at least one optical channel; a fluorescence excitation source; a fluorescence detector; an electronic analog signal amplifier having an input coupled to an output of the fluorescence detector; and an analog-to-digital converter (ADC) having analog input coupled to an output of the analog signal amplifier. The instrument further comprises a quantified automatic gain control (AGC) loop coupled between the signal output of the fluorescence detector and the analog input of the ADC. The AGC loop is configured to apply a determined, numerical gain value to a fluorescence signal for the analog input of the ADC. The instrument also includes a system to scale a digital output of the ADC responsive to the numerical gain value and to provide a digital fluorescence level signal from the scaled digital output.

Abstract: A reaction vessel assembly for use with thermal cyclers is described. The assembly includes a reaction vessel and a casing defining a cavity. In a first configuration, the casing receives the reaction vessel within the cavity, to act as a protective casing for the reaction vessel. In a second configuration, the casing engages with a mouth of the reaction vessel, to close the vessel. In this configuration, the casing may also act as a handle. In preferred embodiments, the reaction vessel is in the form of a capillary tube, and/or may include an integrated collimating lens. Certain embodiments also include an RFID tag.

Abstract: Described is a method for quantifying nucleic acid in a nucleic acid amplification reaction, in which an asymmetric nucleic acid amplification reaction is performed on a sample such that double stranded nucleic acid product is generated in an initial stage of the reaction, and single stranded nucleic acid product is generated in a subsequent stage of the reaction once a limiting primer is exhausted. Relative amounts of double stranded nucleic acid product and single stranded nucleic acid product produced are then detected by means of a melt curve analysis. The ratio of double stranded product peak height to single stranded product peak height may then be used to quantify the amount of starting template based on the ratio of double stranded product peak height to single stranded product peak height.

Abstract: A sample preparation cartridge is described having a lower portion having four integrally-formed leaf springs, and a raised lower base part which is slightly below the level of the leaf springs. An upper portion of the cartridge engages with the lower portion, via four paired pins and holes formed in the lower and upper portions. The upper portion includes three well-shaped openings formed in the central region of the upper portion. Between the upper and lower portions is placed a sheet of sample preparation paper, resting on the leaf springs, which raise the lower face of the paper away from the raised lower base part, leaving a small air gap between the base and the paper. The upper face of the paper is in contact with the well-shaped openings of the upper portion. The user loads a liquid sample into the openings, and the paper wicks the liquid sample away from the loading location, leaving cellular debris in the place of application.

Abstract: An assay for mutations in JAK2 is described. The assay uses selective amplification of mutant alleles with a blocker probe which preferentially hybridizes to wild type alleles. The same probe is then used to detect presence or absence of wild type sequences. It is not necessary to know the specific mutant sequence beforehand.

Abstract: A method for analysing genetic mutations, and in particular single nucleotide polymorphisms (SNPs) and/or somatic mutations, is described, as well as methods for preferentially amplifying one allelic form compared with another form. The methods use an oligonucleotide probe which hybridises to a first allele with a lower melting temperature (Tm) than that with which it hybridises to a second allele, together with amplification primers which flank the oligonucleotide probe binding site and which bind to the sample with a higher Tm than that of the probe and the first allele. An amplification reaction may be carried out at a temperature such that the probe is preferentially hybridised to the second allele, thereby amplifying the first allele. The amplified sequences may be detected using the same probe as acted as the blocking probe during amplification.

Abstract: We describe a thermal cycler comprising a Peltier-type thermoelectric element used for cooling a sample block, and a non-Peltier-type heating device for heating the sample block. The cycler also includes a heat sink connected to the Peltier-type element by a heat pipe, which permits thermal energy to transfer from the Peltier-type element to the heat sink. This configuration operates more efficiently than conventional thermal cyclers which use Peltier-type elements for heating and cooling, and allows a more rapid cycling time as well as operation in a wider range of ambient temperatures. Certain embodiments utilise the Peltier-type element as a thermal gate to reduce thermal loss during heating when the Peltier-type element is switched off.

Abstract: An assembly and related methods are described for the preparation and processing of samples in molecular biology assays and nucleic acid amplification reactions. The assembly allows rapid and easy processing with reduced sample handling compared to known sample preparation apparatus and methods. The assembly comprises a reaction vessel, a sample matrix and a lid. The invention further provides an integrated punch to punch out discs of the sample matrix into the reaction vessel (without handling); thereby reducing the sample handling required to generate the final amplified nucleic acid product.

Abstract: The present invention provides a method of operating a thermal cycler using readable tags (for example, Radio Frequency Identification (RFID)) to simplify the operation and to reduce user interaction requirements. The RFID tag is used to program the thermal cycler unit. This automates process flow and allows single button operation. In general terms, the invention uses RFID tags provided on reaction vessels to identify a particular vessel; while a readable program card contains data associating reaction vessel identities with specific operations (eg, thermal cycling program, detection steps) to be performed on that vessel. The thermal cycler detects the reaction vessel RFID tag, and selects an appropriate operation to perform.

Abstract: A reaction vessel assembly for use with thermal cyclers is described. The assembly includes a reaction vessel and a casing defining a cavity. In a first configuration, the casing receives the reaction vessel within the cavity, to act as a protective casing for the reaction vessel. In a second configuration, the casing engages with a mouth of the reaction vessel, to close the vessel. In this configuration, the casing may also act as a handle. In preferred embodiments, the reaction vessel is in the form of a capillary tube, and/or may include an integrated collimating lens. Certain embodiments also include an RFID tag.

Abstract: A method for analysing genetic mutations, and in particular single nucleotide polymorphisms (SNPs) and/or somatic mutations, is described, as well as methods for preferentially amplifying one allelic form compared with another form. The methods use an oligonucleotide probe which hybridises to a first allele with a lower melting temperature (Tm) than that with which it hybridises to a second allele, together with amplification primers which flank the oligonucleotide probe binding site and which bind to the sample with a higher Tm than that of the probe and the first allele. An amplification reaction may be carried out at a temperature such that the probe is preferentially hybridised to the second allele, thereby amplifying the first allele. The amplified sequences may be detected using the same probe as acted as the blocking probe during amplification.

Abstract: The present invention provides a method of operating a thermal cycler using readable tags (for example, Radio Frequency Identification (RFID)) to simplify the operation and to reduce user interaction requirements. The RFID tag is used to program the thermal cycler unit. This automates process flow and allows single button operation. In general terms, the invention uses RFID tags provided on reaction vessels to identify a particular vessel; while a readable program card contains data associating reaction vessel identities with specific operations (eg, thermal cycling program, detection steps) to be performed on that vessel. The thermal cycler detects the reaction vessel RFID tag, and selects an appropriate operation to perform.

Abstract: Secreted FXYD family proteins are expressed by intestinal mucosa and/or associated tissues to regulate cell production in intestinal crypts in response to tissue damage. Such tissue damage may arise from disease, exposure to injurious chemicals (e.g., due to poisoning, chemotherapy, chemical weapons), or exposure to injurious radiation (e.g., due to nuclear power accidents, radiological weapons, radiation therapy). Because these proteins are secreted in response to epithelial tissue damage, some of them are implicated in tissue repair response or an inflammatory response which prolongs or exacerbates the tissue damage. Examples of these proteins include FXYD 3, FXYD 4, and FXYD 5. Diagnostic methods based upon the role of the FXYD proteins in epithelial tissue damage are disclosed.

Abstract: We describe a thermal cycler comprising a Peltier-type thermoelectric element used for cooling a sample block, and a non-Peltier-type heating device for heating the sample block. The cycler also includes a heat sink connected to the Peltier-type element by a heat pipe, which permits thermal energy to transfer from the Peltier-type element to the heat sink. This configuration operates more efficiently than conventional thermal cyclers which use Peltier-type elements for heating and cooling, and allows a more rapid cycling time as well as operation in a wider range of ambient temperatures. Certain embodiments utilise the Peltier-type element as a thermal gate to reduce thermal loss during heating when the Peltier-type element is switched off.

Abstract: There is provided the use of an inhibitor of phosphate transporter activity for the manufacture of a medicament for the prevention and/or treatment of non-viral damage to an epithelium, or of a condition caused or characterised by such damage. The inhibitor of phosphate transporter activity may optionally be a phosphono-carboxylic acid, or a pharmaceutically acceptable derivative of such an acid. There are also provided methods of treatment using such inhibitors, acids and derivatives.

Abstract: There is provided the use of an inhibitor of phosphate transporter activity for the manufacture of a medicament for the prevention and/or treatment of non-viral damage to an epithelium, or of a condition caused or characterised by such damage. The inhibitor of phosphate transporter activity may optionally be a phosphono-carboxylic acid, or a pharmaceutically acceptable derivative of such an acid. There are also provided methods of treatment using such inhibitors, acids and derivatives.

Abstract: The invention provides a method of determining the degree of similarity between gene expression in a biological sample of interest and that in individual reference samples by use of: i) a nucleic acid probe library representative of a pattern of gene expression in the biological sample of interest; and ii) a plurality of reference samples, each of which is a nucleic acid library representative of a pattern of gene expression in a reference biological sample from which it was derived. The method is effected by: a) forming a first set of immobilised, hybridised products by treating the individual reference samples with the probe library under hybridising conditions (one or other of the reference samples or the probe library being in immobilised form) and removing non-immobilised material; and b) forming a second immobilised product by treating a sample of the free probe library with an immobilised sample of the probe library under hybridising conditions, and removing non-immobilised material.