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US 20120100529A1

(19) United States


(12) Patent Application Publication (10) Pub. No.: US 2012/0100529 A1
(43) Pub. Date:

Fischer et al.

(54)

(75)

BIOLOGICAL SPECIMEN COLLECTION


AND TRANSPORT SYSTEM AND METHODS
OF USE

Inventors:

Gerald W. Fischer, Bethesda, MD

(US); Luke T. Daum, San Antonio,

Apr. 26, 2012

Publication Classi?cation

(51)

Int. Cl.

C12Q 1/70
C12Q 1/68
(52)

(2006.01)
(2006.01)

us. or. .......................................... .. 435/5; 435/6.12

TX (U S)

(57)

(73) Assignee:

Longhorn Vaccines & Diagnostics,


LLC, San Antonio, TX (US)

Disclosed are compositions for isolating populations of


nucleic acids from biological, forensic, and environmental
samples. Also disclosed are methods for using these compo
sitions as one-step formulations for killing pathogens, inac

(21) Appl. No.:

13/332,204

tivating nucleases, and releasing polynucleotides from other

(22)

Filed:

Dec. 20, 2011

ABSTRACT

cellular components Within the sample, and stabilizing the


nucleic acids prior to further processing or assay. The dis

closed compositions safely facilitate rapid sample collection,


and provide extended storage and transport of the samples at

Related US. Application Data

(63)
(60)

Continuation of application No. 12/243,949, ?led on


Oct. 1, 2008, noW Pat. No. 8,084,443.
Provisional application No. 60/976,728, ?led on Oct.

1, 2007.

ambient or elevated temperature Without contamination of the

sample or degradation of the nucleic acids contained therein.


This process particularly facilitates the collection of speci
mens from remote locations, and under conditions previously

considered hostile for preserving the integrity of nucleic acids


released from lysed biological samples Without the need of
refrigeration or freezing prior to molecular analysis.

Patent Application Publication

Apr. 26, 2012 Sheet 1 0f 19

US 2012/0100529 A1

Delta Rn vs Cycle
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Patent Application Publication

Apr. 26, 2012 Sheet 2 0f 19

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US 2012/0100529 A1

Am bion

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Patent Application Publication

Apr. 26, 2012 Sheet 3 0f 19

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US 2012/0100529 A1

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One-Step Solution

23.97

21.37

31.38

RNA Storage Solution

25.35

32.70

32.20

24.39

32.17

32.27

(Ambion)
Water

FIG.3

Patent Application Publication

Apr. 26, 2012 Sheet 4 0f 19

US 2012/0100529 A1

Patent Application Publication

Apr. 26, 2012 Sheet 5 0f 19

US 2012/0100529 A1

95(3)];
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Patent Application Publication

Apr. 26, 2012 Sheet 6 0f 19

US 2012/0100529 A1

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Patent Application Publication

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Apr. 26, 2012 Sheet 7 0f 19

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Patent Application Publication

Apr. 26, 2012 Sheet 8 0f 19

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Patent Application Publication

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Patent Application Publication

Apr. 26, 2012 Sheet 11 0f 19

4% Agarose (HR)
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US 2012/0100529 A1

Patent Application Publication

Apr. 26, 2012 Sheet 12 0f 19

US 2012/0100529 A1

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US 2012/0100529 A1

Patent Application Publication

Apr. 26, 2012 Sheet 14 0f 19

US 2012/0100529 A1

TCEP-Whole influenza A virus

+ TCEP

TChrietscoald
5mM

10mM

25MM

35mM

Molar Concentration

FIG.11

50mM

Patent Application Publication

Apr. 26, 2012 Sheet 15 0f 19

US 2012/0100529 A1

TCEP-H5N1 Avian influenza ssRNA

G2O8ES.

2150

T% P

5mM

10mM

25MM

35mM

Molar Concentration

FIG.12

50mM

Patent Application Publication

Apr. 26, 2012 Sheet 16 0f 19

H5 Avian influenza RNA

33

35

FIG.13A

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35
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,1

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US 2012/0100529 A1

Patent Application Publication

Apr. 26, 2012 Sheet 18 0f 19

US 2012/0100529 A1

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Patent Application Publication

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Apr. 26, 2012 Sheet 19 0f 19

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US 2012/0100529 A1

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