Methods and systems for volume variation modeling in digital pcr

Patent metadata

Field	Value
Publication number	US-2016312274-A1
Application number	US-201615136774-A
Country	US
Kind code	A1
Filing date	Apr 22, 2016
Priority date	Apr 24, 2015
Publication date	Oct 27, 2016
Grant date	—

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.

First claim

What is claimed is:

1 . A method for performing digital polymerase chain reaction (dPCR), 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; determining a model for volume variation of the plurality of partitions; determining a number of partitions including at least one target nucleic acid; and 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.

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

)

]





-

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



(

Assignees

• Life Technologies Corp

Inventors

• Majumdar Nivedita Sumi
• Banerjee Swapnonil

Classifications

• C12Q1/6851 Primary

  Quantitative amplification · CPC title

• G06F17/18

  for evaluating statistical data {, e.g. average values, frequency distributions, probability functions, regression analysis (forecasting specially adapted for a specific administrative, business or logistic context G06Q10/04)} · CPC title

• G16B40/00 Primary

  ICT specially adapted for biostatistics; ICT specially adapted for bioinformatics-related machine learning or data mining, e.g. knowledge discovery or pattern finding · CPC title