IP Library Granted Patent US 7,587,283
Granted Patent B2
US 7,587,283 · App. 11/428,384 · Granted Sep 8, 2009

Growth and baseline identification determination for amplification data

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Quick Facts
Patent No.
US 7,587,283
App. No.
11/428,384
Granted
Sep 8, 2009
Kind
B2
Abstract

Aspects of the present invention describe a method and apparatus for automating analysis of genetic samples. The initial operation receives amplification data associated with at least one spectral species and a genetic sample in each well of a plate. A putative growth signal is identified for the genetic sample from the amplification data forming a baseline region having a start point and an end point along with a growth region. Depending one on one or more characteristics detected in the amplification data forming the baseline region and growth region, aspects of the present invention modifies the baseline region to further identify the putative growth signal.

Claims (23)

1. A computer implemented method of automating analysis of genetic samples, comprising:

receiving amplification data associated with at least one spectral species and a genetic sample in each well of a plate;

identifying a putative growth signal for the genetic sample from the amplification data forming a baseline region having a start point and an end point along with a growth region, the identifying a putative growth signal including

creating the baseline region with a first slope along the amplification data between the start point and the end point, the first slope being significantly less than a second slope along the amplification data associated with the growth region, and

setting a signal threshold level for determining a cycle threshold (C.sub.T) at a predetermined multiple of the variance in the baseline between the starting point and the endpoint; and

modifying the baseline region to further identify the putative growth signal depending on one or more characteristics detected in the amplification data forming the baseline region and growth region.

2. A computer implemented method of automating analysis of genetic samples, comprising:

receiving amplification data associated with at least one spectral species and a genetic sample in each well of a plate;

identifying a putative growth signal for the genetic sample from the amplification data forming a baseline region having a start point and an end point along with a growth region, the identifying a putative growth signal including

creating the baseline region with a first slope along the amplification data between the start point and the end point, the first slope being significantly less than a second slope along the amplification data associated with the growth region, and

setting a signal threshold level for determining a cycle threshold (C.sub.T) at a predetermined multiple of the variance in the baseline between the starting point and the endpoint, wherein the variance from the baseline is selected from a set including: a standard deviation of the baseline from a single well in the plate and a population variance from multiple wells in the plate; and

modifying the baseline region to further identify the putative growth signal depending on one or more characteristics detected in the amplification data forming the baseline region and growth region.

3. The method of claim 2 wherein identifying a putative growth signal having an initial baseline and a growth region further comprises:

measuring a number of zero crossings of the amplification data to determine if the amplification data may correspond to noise; and

obviating anomalous data patterns identified in the amplification data that may interfere with finding a starting point and an endpoint for the baseline and the putative growth signal.

4. The method of claim 3 wherein the anomalous data patterns may include one or more patterns selected from a set of anomalous patterns including: early cycles showing significant excursions in fluorescence values compared to later cycles, significant negative amplitude changes in the growth region of amplification data, significant excursions in fluorescence values during a single cycle, starting point of baseline significantly exceeds variation associated with baseline, and growth region of amplification data does not exceed variation associated with baseline.

5. A computer implemented method of automating analysis of genetic samples, comprising:

receiving amplification data associated with at least one spectral species and a genetic sample in each well of a plate;

identifying a putative growth signal for the genetic sample from the amplification data forming a baseline region having a start point and an end point along with a growth region;

determining whether the amplification data reflects putative noise rather than the putative growth signal associated with growth of the genetic sample during amplification by validating the amplification data associated with the baseline region;

determining whether the putative noise from the amplification data reflects an actual growth signal associated with growth of the genetic sample during amplification rather than putative noise by a further validation of the amplification data associated with the baseline region, wherein determining whether the amplification data reflects an actual growth signal rather than putative noise is based upon whether a product of a maximum amplitude of the amplification data, a second derivative of the amplification data and a baseline end point exceed a predetermined threshold; and

classifying the amplification data from the genetic sample as an actual growth signal responsive to the further validation of the amplification data associated with the baseline region.

6. The method of claim 5 wherein making a final determination of whether or not amplification data is noise or growth signal is determined by a weighted combination of one or more noise detectors and growth detectors derived from the amplification data.

Assignments (5)
CHANGE OF NAME Recorded Feb 26, 2010
From: APPLERA CORPORATION
To: APPLIED BIOSYSTEMS INC.
Reel/Frame 023994/0538 →
MERGER Recorded Feb 26, 2010
From: APPLIED BIOSYSTEMS INC.
To: APPLIED BIOSYSTEMS, LLC
Reel/Frame 023994/0587 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 12, 2008
From: APPLERA CORPORATION
To: CELERA CORPORATION
Reel/Frame 021561/0124 →
CORRECTIVE ASSIGNMENT TO CORRECT THE RECEIVING PARTY WAS INCORRECTLY ENTERED ON THE COVERSHEET PREVIOUSLY RECORDED ON REEL 020619 FRAME 0071. ASSIGNOR(S) HEREBY CONFIRMS THE RECEIVING PARTY. Recorded Mar 13, 2008
From: LEONG, HARRISON M.F., PH.D
To: APPLERA CORPORATION
Reel/Frame 020649/0765 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 7, 2008
From: LEONG, HARRISON M.F., PH.D
To: NETWORK APPLIANCE, INC.
Reel/Frame 020619/0071 →