# US 2019/0376129 A1 · Nucleic acid amplification and detection apparatus and method > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0376129 A1 - Document ID: US20190376129A1 - Status: Published application - Kind: A1 - Application number: 16/548,643 - Filed: 2019-08-22 - Published: 2019-12-12 - Earliest claimed: 2012-02-03 - Assignees: Axxin Pty Ltd - Applicants: Axxin Pty Ltd - Inventors: William Robb HOPPER - Web page: https://patentgrep.com/patent/US20190376129A1 ## CPC - C12Q 1/6851: Quantitative amplification - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples} - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE ## Abstract A nucleic acid amplification and detection apparatus, including: a support configured to receive a plurality of reaction vessels containing respective samples of one or more nucleic acids to be amplified, the support being rotatable about an axis of rotation and the reaction vessels being received in the support at respective receiving locations distributed about the axis of rotation; a temperature control component thermally coupled to the support and configured to control the temperature of the support in order to amplify the nucleic acids contained in the reaction vessels while received in the support; one or more measurement components configured to measure one or more characteristics of the nucleic acids within the reaction vessels at respective measurement locations distributed about the axis of rotation; an actuator coupled to the support and configured to rotate the support about the axis of rotation; and a sample position controller coupled to the actuator and being configured to rotate the support about the axis of rotation so as to position a selected one of the plurality of reaction vessels to a selected one of the measurement locations to allow a corresponding one of the measurement components to perform a corresponding measurement on the corresponding sample. ## Claims ### Claim 1 A nucleic acid amplification and detection method, including: receiving signal data representing assay measurements of a sample containing at least one nucleic acid at respective times during amplification of the at least one nucleic acid; processing said signal data to generate second derivative data representing values of a second derivative of said signal data with respect to time; processing said second derivative data to determine whether at least one of the second derivative values exceeds a predetermined second derivative threshold value; processing said second derivative data to determine a crossover time at which the second derivative of said signal data crosses zero after having exceeded the second derivative threshold value; processing said signal data to generate first derivative data representing a first derivative of said signal data with respect to time at the crossover time; generating assay result data representing a result of the assay, the result being determined on the basis of: (i) whether the predetermined second derivative threshold value was exceeded, and (ii) the first derivative of said signal with respect to time at the crossover time; and outputting the assay result data. ### Claim 2 (depends on 1) The method of claim 1, wherein the assay measurements are fluorescence measurements during amplification of the at least one nucleic acid. ### Claim 3 (depends on 1) The method of claim 1, wherein receiving the signal data representing assay measurements comprises receiving signal data acquired using multiple non-overlapping wavelengths for amplification of the at least one nucleic acid. ### Claim 4 (depends on 1) The method of claim 1, wherein the assay measurements are taken using a signal amplification temperature in the range of about 35-65° C. ### Claim 5 (depends on 1) The method of claim 1, wherein the result is determined on the basis of whether a positive going peak in the second derivative is present. ### Claim 6 (depends on 5) The method of claim 5, wherein the result is determined on the basis of whether the positive going peak in the second derivative has a height above a given threshold. ### Claim 7 (depends on 6) The method of claim 6, wherein the result is determined on the basis of whether the positive going peak in the second derivative has a width or integrated area under its curve that exceeds a corresponding predetermined threshold value. ### Claim 8 (depends on 1) The method of claim 1, further comprising determining whether the result of the assay is asserted or not asserted based on a positive peak height of the second derivative curve, an area under the second derivative curve for the positive and negative going curves, or a width of a peak in the second derivative curve. ### Claim 9 (depends on 1) The method of claim 1, further comprising determining whether the result of the assay is asserted or not asserted based on whether: (i) at least one of the second derivative values exceeds a predetermined positive second derivative threshold value; (ii) the second derivative values cross zero after having exceeded the second derivative threshold value of (i), the time at which this occurs being referred to as the zero crossing time; and (iii) a measure of a width or integrated second derivative values exceeds a further corresponding predetermined threshold value. ### Claim 10 (depends on 9) The method of claim 9, wherein the width is a full width at half-maximum (FWHM). ### Claim 11 (depends on 9) The method of claim 9, wherein the integrated second derivative values are equivalent to the first derivative values at the zero crossing time of (ii). ### Claim 12 (depends on 1) The method of claim 1, wherein amplification of the at least one nucleic acid is for one or more isothermal nucleic acid reactions. ### Claim 13 A nucleic acid amplification and detection method, including: receiving signal data representing assay measurements of a sample containing at least one nucleic acid at respective times during amplification of the at least one nucleic acid; processing said signal data to generate second derivative data representing values of a second derivative of said signal data with respect to time; processing said second derivative data to determine whether at least one of the second derivative values exceeds a predetermined second derivative threshold value; processing said second derivative data to determine whether the second derivative of said signal data crosses zero after having exceeded the second derivative threshold value; processing said second derivative data to determine which of the second derivative values are positive; determining a width or integrated area of the positive second derivative values; comparing the width or integrated area of the positive second derivative values with a further predetermined threshold value; generating assay result data representing a result of the assay, the result being determined on the basis of: (i) whether the predetermined second derivative threshold value was exceeded, (ii) whether the second derivative of said signal crosses zero after having exceeded the second derivative threshold value, and (iii) whether the width or integrated area of the positive second derivative values exceeds the further predetermined threshold value; and outputting the assay result data. ### Claim 14 (depends on 13) The method of claim 13, wherein the result is determined on the basis of whether a positive going peak in the second derivative is present. ### Claim 15 (depends on 14) The method of claim 14, wherein the result is determined on the basis of whether the positive going peak in the second derivative has a height above a given threshold. ### Claim 16 (depends on 15) The method of claim 15, wherein the result is determined on the basis of whether the positive going peak in the second derivative has a width or integrated area under its curve that exceeds a corresponding predetermined threshold value. ### Claim 17 (depends on 13) The method of claim 13, further comprising determining whether the result of the assay is asserted or not asserted based on a positive peak height of the second derivative curve, an area under the second derivative curve for the positive and negative going curves, or a width of a peak in the second derivative curve. ### Claim 18 (depends on 13) The method of claim 13, wherein amplification of the at least one nucleic acid is for one or more isothermal nucleic acid reactions. ### Claim 19 (depends on 13) The method of claim 13, wherein the assay measurements are taken using a signal amplification temperature in the range of about 35-65° C. ### Claim 20 (depends on 13) The method of claim 13, wherein receiving the signal data representing assay measurements comprises receiving signal data acquired using multiple non-overlapping wavelengths for amplification of the at least one nucleic acid.