# US 2013/0157276 A1 · Device and method for conducting direct quantitive real time pcr > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2013/0157276 A1 - Document ID: US20130157276A1 - Status: Published application - Kind: A1 - Application number: 13/817,899 - Filed: 2010-09-24 - Published: 2013-06-20 - Earliest claimed: 2010-09-24 - Assignees: Alphahelix Molecular Diagnostics AB - Inventors: Lars Edvinsson; Magnus Molin - Web page: https://patentgrep.com/patent/US20130157276A1 ## CPC - C12Q 1/686: Polymerase chain reaction [PCR] - B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples} - B01L 7/525: {with physical movement of samples between temperature zones} - G01N 21/07: Centrifugal type cuvettes (G01N21/09 takes precedence) - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept - B01L 2200/14: Process control and prevention of errors - B01L 2300/0654: Lenses; Optical fibres - B01L 2400/0409: centrifugal forces - G01N 2021/513: {Cuvettes for scattering measurements} - G01N 2021/6491: {Measuring fluorescence and transmission; Correcting inner filter effect} - 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 - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - 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) ## Abstract A method and device for performing direct quantitative real time PCR in a crude sample (200), wherein said sample is subjected to a centrifugal force sufficient to separate components of the sample into a supernatant and a pellet, and wherein at least one light source (204) and at least one detector (205) are positioned so that the excitation light impinges on the sample in a position above said pellet, and said detector detects light emitted from a position above said pellet. ## Claims ### Claim 1 A device for performing direct quantitative real time PCR, said device comprising a rotor for receiving one or more elongate containers having an opening for receiving one or more samples, a distal end, and a longitudinal axis, rotating said containers along a circular path and subjecting said samples in said containers to a centrifugal force, at least one light source for subjecting the sample to excitation light, and at least one detector for detection light emitted from the sample, wherein said rotor holds said elongate containers in a position where the longitudinal axis of each container is parallel to the radius of said rotor; at least one light source and said at least one detector are positioned in relation to the rotor so that the excitation light impinges on the container in a position between the distal end of the container and the opening of the container, and said detector detects light emitted from the sample from a position between the distal end of the container and the opening of the container. ### Claim 2 (depends on 1) The device according to claim 1, wherein said centrifugal force is sufficient to separate the components of the sample into a supernatant and a pellet. ### Claim 3 (canceled) ### Claim 4 (depends on 1) The device according to claim 1, wherein said centrifugal force is at least about 1000×g, preferably at least about 2000×g, more preferably at least about 3000×g. ### Claim 5 (depends on 1) The device according to claim 1, wherein said at least one light source and said at least one detector are positioned in relation to the rotor so that the excitation light impinges on the container in a position above the pellet formed in the container, and said detector detects light emitted from the sample from a position above the pellet formed in the container. ### Claim 6 (depends on 1) The device according to claim 1, wherein at least one of said light source and said detector is/are movably arranged in relation to the circular path taken by the elongate containers. ### Claim 7 (depends on 6) The device according to claim 6, wherein at least one of said light source and said detector is/are movably arranged to be moved along or parallel to the radius of the circular path taken by the containers. ### Claim 8 (depends on 1) The device according to claim 1, wherein the device further comprises a motor operatively connected to the rotor, a control unit, and a user interface. ### Claim 9 (depends on 1) The device according to claim 1, wherein said device further comprises means for measuring the absorption and/or diffusion of light in the sample. ### Claim 10 (depends on 1) The device according to claim 1, wherein the device further comprises an outer cover and an inner cover wherein said inner cover is adapted to be at least partially opened during operation of the rotor without opening the outer cover. ### Claim 11 A method for performing direct quantitative real time PCR on a sample comprising the steps of placing said sample in an elongate container, subjecting said sample in said container to a centrifugal force, subjecting the sample to excitation light from a light source, detecting light emitted from the sample using a detector, wherein said elongate containers are subjected to a centrifugal force in a rotor which holds the containers in a position where the longitudinal axis of each container is parallel to the radius of said rotor; wherein at least one light source and said at least one detector are positioned in relation to the rotor so that the excitation light impinges on the container in a position between the distal end of the container and the opening of the container, and said detector detects light emitted from the sample from a position between the distal end of the container and the opening of the container. ### Claim 12 (depends on 11) The method according to claim 11, wherein said sample is subjected to a centrifugal force sufficient to separate components of the sample into a supernatant and a pellet. ### Claim 13 (canceled) ### Claim 14 (depends on 11) The method according to claim 11, wherein said centrifugal force is at least about 1000×g, preferably at least about 2000×g, more preferably at least about 3000×g. ### Claim 15 (depends on 11) The method according to claim 11, wherein said at least one light source and said at least one detector are positioned in relation to the rotor so that the excitation light from said at least one light source impinges on the container in a position above the pellet formed in the container, and said at least one detector detects light emitted from the sample from a position above the pellet formed in the container. ### Claim 16 A method for performing direct quantitative real time PCR on a sample, comprising: placing said sample in an elongate container, subjecting said sample in said container to a centrifugal force, subjecting the sample to excitation light from a light source, detecting light emitted from the sample using a detector, wherein said elongate containers are subjected to a centrifugal force in a rotor which holds the containers in a position where the longitudinal axis of each container is parallel to the radius of said rotor; wherein at least one light source and said at least one detector are positioned in relation to the rotor so that the excitation light impinges on the container in a position between the distal end of the container and the opening of the container, and said detector detects light emitted from the sample from a position between the distal end of the container and the opening of the container, and wherein the interface between the pellet and the supernatant is detected, and said at least one light source and/or said at least one detector are moved in relation to the rotor so that the excitation light impinges on the container in a position above the pellet formed in the container, and said detector detects light emitted from the sample from a position above the pellet formed in the container. ### Claim 17 (depends on 16) The method according to claim 16, wherein at least one of said light source and said detector is/are moved in relation to the circular path taken by the elongate containers. ### Claim 18 (depends on 16) The method according to claim 16, wherein at least one of said light source and said detector is/are movably arranged to be moved along or parallel to the radius of the circular path taken by the containers. ### Claim 19 (depends on 11) The method according to claim 11, wherein the absorbance of the sample is measured intermittently or continuously, and a measured change in absorbance is used to reduce and compensate for the influence of absorption on the fluorescence. ### Claim 20 (depends on 19) The method according to claim 19, wherein the absorbance is measured in the supernatant. ### Claim 21 (depends on 11) The method according to claim 11, wherein the diffusion and/or reflectance of the sample is measured intermittently or continuously, and a measured change in diffusion and/or reflectance is used to reduce and compensate for the influence of absorption on the fluorescence. ### Claim 22 (depends on 21) The method according to claim 21, wherein the diffusion and/or reflectance is measured in the supernatant.