# US 8,569,020 B2 · Method for simultaneously performing multiple amplification reactions

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 8,569,020 B2
- Document ID: US8569020B2
- Status: Granted
- Kind: B2
- Application number: 12/241,350
- Filed: 2008-09-30
- Published: 2009-01-29
- Granted: 2013-10-29
- Earliest claimed: 1998-05-01
- Assignees: Gen-Probe Incorporated
- Applicants: Kelly G. Ammann; Ralph E. Burns; Ernest V. Hansberry; Glenn A. Horner; Cheryl A. Jakub; John E. Kling; Donald J. Nieglos; Robert E. Schneider; Robert J. Smith
- Inventors: Kelly G. Ammann; Ralph E. Burns; Ernest V. Hansberry; Glenn A. Horner; Cheryl A. Jakub; John E. Kling; Donald J. Nieglos; Robert E. Schneider; Robert J. Smith
- Web page: https://patentgrep.com/patent/US8569020B2

## CPC

- C12Q 1/6834: Enzymatic or biochemical coupling of nucleic acids to a solid phase
- B01F 29/10: with receptacles rotated about two different axes, e.g. receptacles having planetary motion
- B01F 29/30: Mixing the contents of individual packages or containers, e.g. by rotating tins or bottles
- B01F 29/322: of two or more containers supported for simultaneous mixing, e.g. for bottles in crates
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- B01L 7/5255: {by moving sample containers}
- B03C 1/282: {with associated accumulation indicator, e.g. Hall sensor}
- B03C 1/288: {disposed at the outer circumference of a recipient}
- B03C 1/30: Combinations with other devices, not otherwise provided for
- C12Q 1/6813: Hybridisation assays
- C12Q 1/6832: Enhancement of hybridisation reaction
- G01N 35/025: {having a carousel or turntable for reaction cells or cuvettes}
- B01F 29/40354: {arranged for planetary motion}
- B01L 9/06: Test-tube stands; Test-tube holders
- B01L 2200/147: Employing temperature sensors
- B01L 2300/1822: using Peltier elements
- B01L 2300/1827: using resistive heater
- B03C 2201/26: for use in medical or biological applications
- C12Q 2537/10: the purpose or use of
- G01N 35/0098: {involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation (magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C)}
- G01N 35/0099: {comprising robots or similar manipulators (robots per se B25J)}
- G01N 35/1065: {Multiple transfer devices}
- G01N 2035/00356: {Holding samples at elevated temperature (incubation)}
- G01N 2035/00524: {Mixing by agitating sample carrier}
- G01N 2035/0437: {Cleaning cuvettes or reaction vessels}
- G01N 2035/0455: {Coaxial carousels}
- G01N 2035/103: {using disposable tips}
- Y10T 29/53657: to apply or remove a resilient article [e.g., tube, sleeve, etc.]
- Y10T 29/53683: Spreading parts apart or separating them from face to face engagement
- Y10T 29/53843: Tube, sleeve, or ferrule inserting or removing
- Y10T 156/1105: Delaminating process responsive to feed or shape at delamination
- Y10T 156/1906: Delaminating means responsive to feed or shape at delamination
- Y10T 436/11: Automated chemical analysis
- Y10T 436/111666: Utilizing a centrifuge or compartmented rotor
- Y10T 436/113332: with conveyance of sample along a test line in a container or rack
- Y10T 436/114998: with treatment or replacement of aspirator element [e.g., cleaning, etc.]
- Y10T 436/119163: with aspirator of claimed structure
- Y10T 436/2575: Volumetric liquid transfer
- 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
- B01F: MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING (mixing paints B44D3/06)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B03C: MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS (separating isotopes B01D59/00; combinations of magnetic or electrostatic separation with separation of solids by other means B03B, B07B)
- 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)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

An automated method for performing multiple nucleic acid amplification procedures in multiple receptacles of a set of receptacles contained in an analyzer.

## Claims

### Claim 1

A method for detecting the presence of a nucleic acid in each of multiple samples, the method comprising performing within a housing of an analyzer the automated steps of:
(a) contacting each of multiple samples containing a nucleic acid with a solid support, such that a complex comprising the nucleic acid and the solid support is formed in each of the samples, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex formed in each of the samples is suspended in a fluid component of the sample;
(b) after step (a), subjecting the complex in each of the samples to a magnetic field;
(c) during step (b), purifying the nucleic acid in each of the samples by accessing and removing at least a portion of the fluid component of each of the samples from the complex;
(d) forming multiple mixtures, each of the mixtures comprising the purified nucleic acid of one of the samples of step (c) and reagents for performing a nucleic acid amplification, wherein the mixtures are separately formed in multiple receptacles of a set of receptacles;
(e) moving the set of receptacles to a temperature-controlled environment;
(f) in each of the multiple receptacles, performing nucleic acid amplification, whereby amplification products are enzymatically synthesized, each of the amplification products comprising (i) a target sequence contained in the nucleic acid or (ii) the complement of the target sequence;
(g) in each of the multiple receptacles, selectively hybridizing a detectably labeled probe to an amplification product that is one of the amplification products synthesized in step (f), thereby forming a hybrid that comprises the probe and the amplification product, wherein the amplification product comprises the target sequence or its complement, and wherein the probe hybridizes to the target sequence or its complement in the amplification product; and
(h) in each of the multiple receptacles, detecting the presence of the hybrid, wherein the presence of the hybrid in each of the multiple receptacles is an indication of the presence of the nucleic acid in each of the samples,
wherein the method is performed without human intervention.

### Claim 2 (depends on 1)

The method of claim 1, further comprising performing within the housing of the analyzer, prior to step (h) and after step (g), a step of providing a selection reagent to each of the multiple receptacles, whereby a label of an unhybridized, detectably labeled probe in each of the multiple receptacles is selectively hydrolyzed, wherein the label is an acridinium ester.

### Claim 3 (depends on 1)

The method of claim 1, further comprising performing within the housing of the analyzer an automated step of washing the solid support of each of the samples one or more times after step (c) and prior to step (d).

### Claim 4 (depends on 1)

The method of claim 1, wherein each receptacle of the set of receptacles comprises a cylindrical tube.

### Claim 5 (depends on 1)

The method of claim 1, wherein the set of receptacles is an integrally formed set of receptacles.

### Claim 6 (depends on 1)

The method of claim 1, wherein each of the amplification products consists of a nucleic acid sequence having the target sequence or its complement.

### Claim 7 (depends on 1)

The method of claim 1, wherein the amplification products include nucleic acid sequences having a sequence that is the same as the target sequence and nucleic acid sequences having a sequence that is the complement of the target sequence.

### Claim 8 (depends on 1)

The method of claim 1, wherein step (f) is carried out in a temperature- controlled incubator maintained at a substantially uniform temperature.

### Claim 9 (depends on 1)

The method of claim 1, wherein step (f) is performed in a temperature-controlled chamber of a first incubator, and wherein step (g) is performed in a temperature-controlled chamber of a second incubator, the first and second incubators being in a spaced-apart relationship.

### Claim 10 (depends on 1)

The method of claim 1, wherein the probe is detectably labeled with a fluorescent or chemiluminescent label.

### Claim 11 (depends on 1)

The method of claim 1, wherein the hybrid formed in each of the multiple receptacles is in solution in steps (g) and (h).

### Claim 12 (depends on 1)

The method of claim 1, wherein the method is performed at multiple locations of a processing deck contained within the housing.

### Claim 13 (depends on 1)

The method of claim 1, wherein the housing remains closed during the method.

### Claim 14 (depends on 5)

The method of claim 5, wherein the integrally formed receptacles are arranged in a linear array.

### Claim 15

A method for detecting the presence of a nucleic acid in each of multiple samples, the method comprising performing within a housing of an analyzer the automated steps of:
(a) contacting each of multiple samples containing a nucleic acid with a solid support, such that a complex comprising the nucleic acid and the solid support is formed in each of the samples, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex formed in each of the samples is suspended in a fluid component of the sample;
(b) after step (a), subjecting the complex in each of the samples to a magnetic field;
(c) during step (b), purifying the nucleic acid in each of the samples by contacting the fluid component of each of the samples with a fluid aspirator and aspirating at least a portion of the fluid component from the complex;
(d) forming multiple mixtures, each of the mixtures comprising the purified nucleic acid of one of the samples of step (c) and reagents for performing a nucleic acid amplification, wherein the mixtures are separately formed in multiple receptacles of a set of receptacles;
(e) moving the set of receptacles to a temperature-controlled environment;
(f) in each of the multiple receptacles, performing nucleic acid amplification, amplification products are enzymatically synthesized, each of the amplification products comprising (i) a target sequence contained in the nucleic acid or (ii) the complement of the target sequence;
(g) in each of the multiple receptacles, selectively hybridizing a detectably labeled probe to an amplification product that is one of the amplification products synthesized in step (f), thereby forming a hybrid that comprises the probe and the amplification product, wherein the amplification product comprises the target sequence or its complement, wherein the probe hybridizes to the target sequence or its complement in the amplification product, and wherein the hybrid is formed in solution; and
(h) in each of the multiple receptacles, detecting the presence of the hybrid while the hybrid is in solution, wherein the presence of the hybrid in each of the multiple receptacles is an indication of the presence of the nucleic acid in each of the samples,
wherein the method is performed without human intervention.

### Claim 16 (depends on 15)

The method of claim 15, further comprising performing within the housing of the analyzer, prior to step (h) and after step (g), a step of providing a selection reagent to each of the multiple receptacles, whereby a label of an unhybridized, detectably labeled probe in each of the multiple receptacles is selectively hydrolyzed, wherein the label is an acridinium ester.

### Claim 17 (depends on 15)

The method of claim 15, wherein step (f) is performed in a temperature-controlled chamber of a first incubator, and wherein step (g) is performed in a temperature-controlled chamber of a second incubator, the first and second incubators being in a spaced-apart relationship.

### Claim 18 (depends on 15)

The method of claim 15 further comprising performing within the housing of the analyzer an automated step of washing the solid support of each of the samples one or more times after step (c) and prior to step (d).

### Claim 19 (depends on 15)

The method of claim 15, wherein each receptacle of the set of receptacles comprises a cylindrical tube.

### Claim 20 (depends on 15)

The method of claim 15, wherein the set of receptacles is an integrally formed set of receptacles.

### Claim 21 (depends on 20)

The method of claim 20, wherein the integrally formed receptacles are arranged in a linear array.

### Claim 22 (depends on 15)

The method of claim 15, wherein the fluid aspirator comprises an aspirator tube in frictional engagement with a contamination-limiting element configured to permit the passage of the fluid component of the specimen therethrough, and wherein the contamination- limiting element contacts the fluid component of the specimen in step (c).

### Claim 23 (depends on 22)

The method of claim 22, wherein the contamination-limiting element is a tiplet.

### Claim 24 (depends on 15)

The method of claim 15, wherein each of the amplification products consists of a nucleic acid sequence having the target sequence or its complement.

### Claim 25 (depends on 15)

The method of claim 15, wherein the amplification products include nucleic acid sequences having a sequence that is the same as the target sequence and nucleic acid sequences having a sequence that is the complement of the target sequence.

### Claim 26 (depends on 15)

The method of claim 15, wherein step (f) is carried out in a temperature-controlled incubator maintained at a substantially uniform temperature.

### Claim 27 (depends on 15)

The method of claim 15, wherein the probe is detectably labeled with a fluorescent or chemiluminescent label.

### Claim 28 (depends on 15)

The method of claim 15, wherein the method is performed at multiple locations of a processing deck contained within the housing.

### Claim 29 (depends on 15)

The method of claim 15, wherein the housing remains closed during the method.

### Claim 30 (depends on 15)

The method of claim 15, wherein the hybird formed in each of the multiple receptacles is in solution.