US10385385B2ActiveUtilityA1
Methods and systems for volume variation modeling in digital PCR
Est. expiryApr 24, 2035(~8.8 yrs left)· nominal 20-yr term from priority
G06F 17/18C12Q 1/6851G16B 40/00
34
PatentIndex Score
0
Cited by
6
References
18
Claims
Abstract
A method for performing digital polymerase chain reaction (dPCR) is provided. The method includes partitioning a biological sample volume including a plurality of target nucleic acids into a plurality of partitions, where at least one partition includes at least one target nucleic acid. The method further includes determining a model for volume variation of the plurality of partitions and determining a number of partitions including at least one target nucleic acid. The method includes generating a concentration of target nucleic acids in the biological sample based on the model for volume variation and the fraction of partitions including at least one target nucleic acid.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1. A method for performing digital polymerase chain reaction (dPCR) in a dPCR biological analysis system including a thermal cycler, a processor, a detector, and a display, the method comprising:
partitioning a biological sample volume including a plurality of target nucleic acids into a plurality of partitions, wherein at least one partition includes at least one target nucleic acid;
amplifying, with the thermal cycler, the target nucleic acids in the plurality of partitions;
generating, by the processor, a model for volume variation of the plurality of partitions based on a normal distribution of effective load volumes of the biological sample volume in the plurality of partitions;
detecting, by the detector, the amplified target nucleic acids to determine a number of partitions including at least one target nucleic acid;
calculating, by the processor, a concentration of target nucleic acids in the biological sample based on the model for volume variation and the number of partitions including at least one target nucleic acid; and
displaying, on the display, the concentration of target nucleic acids in the biological sample.
2. The method of claim 1 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation:
C
=
v
0
-
v
0
2
+
2
σ
2
ln
P
(
neg
)
σ
2
.
3. The method of claim 1 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation:
P
(
neg
)
=
erfc
[
-
1
2
(
v
0
σ
-
C
σ
)
]
erfc
[
-
1
2
(
v
0
σ
)
]
e
-
Cv
0
+
1
2
σ
2
C
2
.
4. The method of claim 1 , further comprising:
amplifying the target nucleic acids to determine the number of partitions including at least one target nucleic acid.
5. The method of claim 1 , wherein the model for volume variation is:
P
(
neg
)
=
erfc
[
-
1
2
(
v
0
σ
-
C
σ
)
]
erfc
[
-
1
2
(
v
0
σ
)
]
e
-
Cv
0
+
1
2
σ
2
C
2
.
6. The method of claim 1 , wherein the model for volume variation is:
P (neg)=exp(1/2σ 2 C 2 −C v 0 ).
7. The method of claim 1 , wherein the plurality of partitions is a plurality of reaction sites.
8. The method of claim 1 , wherein the plurality of partitions is a plurality of throughholes.
9. The method of claim 1 , wherein the plurality of partitions is a plurality of droplets.
10. A system for performing digital polymerase chain reaction (dPCR), the system comprising:
a device configured to partition a biological sample volume including a plurality of target nucleic acids into a plurality of partitions, wherein at least one partition includes at least one target nucleic acid;
a thermal cycler to amplify the plurality of target nucleic acids;
a detector for detecting the number of partitions including at least one target nucleic acid;
a memory; and
a processor configured to:
generate a concentration of target nucleic acids in the biological sample based on a model for volume variation and the number of partitions including at least one target nucleic acid, wherein the model is based on a normal distribution of effective load volume of each of the biological sample volume in the plurality of partitions; and
a display to display the concentration of target nucleic acids in the biological sample.
11. The system of claim 10 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation:
C
=
v
0
-
v
0
2
+
2
σ
2
ln
P
(
neg
)
σ
2
.
12. The system of claim 10 , wherein the concentration of target nucleic acids in the biological sample is determined by using the equation:
P
(
neg
)
=
erfc
[
-
1
2
(
v
0
σ
-
C
σ
)
]
erfc
[
-
1
2
(
v
0
σ
)
]
e
-
Cv
0
+
1
2
σ
2
C
2
.
13. The system of claim 10 , further comprising:
an amplification apparatus configured to amplify the target nucleic acids to determine the number of partitions including at least one target nucleic acid.
14. The system of claim 10 , wherein the model for volume variation is:
P
(
neg
)
=
erfc
[
-
1
2
(
v
0
σ
-
C
σ
)
]
erfc
[
-
1
2
(
v
0
σ
)
]
e
-
Cv
0
+
1
2
σ
2
C
2
.
15. The system of claim 10 , wherein the model for volume variation is:
P (neg)=exp(1/2σ 2 C 2 −C v 0 ).
16. The system of claim 10 , wherein the plurality of partitions is a plurality of reaction sites.
17. The system of claim 10 , wherein the plurality of partitions is a plurality of throughholes.
18. The system of claim 10 , wherein the plurality of partitions is a plurality of droplets.Join the waitlist — get patent alerts
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