Granted Patent B2

US 12,584,161 · App. 17/521,350 · Granted Mar 24, 2026

Inventors: Jianbing Qin (Omaha, NE); Bradford A. Hunsley (Papillion, NE)

Assignee: STRECK LLC

C12Q1/6806

Abstract

Claims (37)

1 . A composition comprising:

(a) a urine sample;

(b) a nuclease inhibitor present in a concentration of at least 0.5% of the composition;

(c) a preservative present in a concentration of from about 0.2% to about 4% of the composition;

(d) a formaldehyde quencher present in a concentration of at least 0.01%, wherein a ratio of nuclease inhibitor to preservative is about 9:10 or about 9:20.

2 . A specimen container comprising a urine preservative composition for maintaining structural integrity of nucleated blood cells and cell-free nucleic acids in a urine sample comprising:

(a) a nuclease inhibitor present in an amount of from about 100 to about 300 g/L of the composition;

(b) a preservative;

(c) a formaldehyde quencher, wherein the ratio of nuclease inhibitor to preservative is about 9:10 or about 9:20.

3 . The specimen container of claim 2 , wherein the urine preservative composition is present in an amount of about 1 to 15 mL.

4 . The specimen container of claim 2 , wherein the nuclease inhibitor is ethylene glycol tetraccetic acid (EGTA), pepstatin, K 3 EDTA, phosphoramidone, leupeptin, aproptinin, bestatin, proteinase inhibitor E 64 (E-64), 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), or a combination thereof.

5 . The specimen container of claim 2 , wherein the formaldehyde quencher is glycine, Tris(hydroxymethyl)aminomethane (TRIS), urea, allantoin, sulfites, or a combination thereof.

6 . The specimen container of claim 2 , wherein preservative comprises imidazolidinyl urea (IDU), diazolidinyl urea (DU), 2-bromo-2nitropropane01,3,-diol, 5-hydroxymethoxymethyl-1-aza-3,7-dioxabicyclo (3.3.0) octane and 5-hydroxymethyl-1-aza-3,7-dioxabicyclo (3.3.0) octaine and 5-hydroxypoly[methyleneoxy]methyl-1-aza-3,7-dioxabicyclo (3.3.0) octaine, a bicyclic oxazolidine, DMDM hydantoin, sodium hydroxymethylglycinate, hexamethylenetetramine chloroallyl chlrodie (Quaternium 15), a biocide, a water-soluble zinc salt, or a combination thereof.

7 . The specimen container of claim 2 , wherein the preservative comprises IDU, DU, or a combination thereof and the formaldehyde quencher comprises glycine.

8 . The specimen container of claim 2 , wherein the ratio of nuclease inhibitor to preservative is about 9:10 and a ratio of nuclease inhibitor to formaldehyde quencher is about 9:1.

9 . The specimen container of claim 2 , wherein the urine preservative composition is provided in an amount for a single unit dosage.

10 . The composition of claim 1 , wherein the urine sample is diluted at a ratio in a range of 5 parts urine sample to 1 part urine preservative composition to 20 parts urine sample to 1 part urine preservative composition.

11 . The composition of claim 10 , wherein the urine sample is diluted at a ratio in a range of 5 parts urine sample to 1 part urine preservative composition.

12 . The composition of claim 8 , wherein preservative comprises imidazolidinyl urea (IDU), diazolidinyl urea (DU), 2-bromo-2nitropropane01,3,-diol, 5-hydroxymethoxymethyl-1-aza-3,7-dioxabicyclo (3.3.0) octane and 5-hydroxymethyl-1-aza-3,7-dioxabicyclo (3.3.0) octaine and 5-hydroxypoly[methyleneoxy]methyl-1-aza-3,7-dioxabicyclo (3.3.0) octaine, a bicyclic oxazolidine, DMDM hydantoin, sodium hydroxymethylglycinate, hexamethylenetetramine chloroallyl chloride (Quaternium 15), a biocide, a water-soluble zinc salt, or a combination thereof.

13 . A method for preserving a urine sample comprising the steps of:

(a) providing the specimen container of claim 2 comprising a predetermined amount of the urine preservative composition;

(b) contacting the urine preservative composition with a urine sample;

(c) sealing the specimen container with the urine sample and the urine preservative composition contained within it; and

wherein the urine sample, upon being preserved, is capable of analysis after a period at least 168 hours has elapsed.

14 . The method of claim 13 , wherein the urine preservative composition comprises: imidazolidinyl urea (IDU), diazolidinyl urea (DU) or a combination thereof as the preservative, and glycine as the formaldehyde quencher.

15 . The method of claim 13 , wherein the preservative includes K 3 EDTA in an amount of at least five times what would be necessary for inhibition of nuclease activity in a blood sample.

16 . The method of claim 13 , wherein the urine preservative composition is present in an amount of about 1 to about 20 percent by volume of the specimen container.

17 . The method of claim 13 , wherein the specimen containers comprises a single unit dose of the urine preservative.

18 . The method of claim 13 , wherein the urine preservative is directly contacted with urine sample or the urine sample is directly contacted with the urine preservative within the specimen container.

19 . The method of claim 13 , wherein the shipping temperature of the sealed specimen container is within a range from about 6° C. to about 37° C.

20 . The method of claim 13 , wherein the sample exhibits a stability for at least about 7 days post treatment.

21 . The method of claim 13 , further comprising analyzing the urine sample, wherein the step of analysis is performed 1, 2, 4, or 7 days after the specimen is obtained.

22 . The method of claim 21 , where the analysis of the urine sample includes nucleic acids from the preserved urine sample and analyzing the isolated nucleic acids to identify a pathological characteristic.

23 . The method of claim 13 , wherein the method is performed in the absence of refrigeration.

24 . The method of claim 13 , wherein the urine includes one or more of cell free nucleic acids, cellular nucleic acids, DNA, circulating tumor DNA (ctDNA), RNA, microRNA (miRNA), or messenger RNA (m RNA).

25 . The method of claim 13 , wherein nucleic acid analysis is performed on one or more of a cell free DNA, circulating tumor DNA (ctDNA), cell free RNA, mRNA, or miRNA from within the urine.

26 . The method of claim 13 , wherein the specimen container contains preservative composition present in an amount of about 1 to about 15 ml.

Assignments (3)

Continuity (3)

References Cited (400)