Reagent for analyzing solid components in urine and method for analyzing solid components by employing the same

• US 5,891,733 A
• Filed: 10/20/1995
• Issued: 04/06/1999
• Est. Priority Date: 10/20/1994
• Status: Expired due to Term

• Current Assignee: Sysmex Corporation
• Original Assignee: Toa Medical Electronics Co., Ltd.
• Inventors: Inoue, Junya
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US08/545,939
• Time in Patent Office: 1,264 Days
• Field of Search: 436/8, 436/10, 436/16, 436/17, 436/18, 436/43, 436/52, 436/63, 436/172, 435/6
• US Class Current: 436/63
• CPC Class Codes: G01N 15/12 by observing changes in resistance or impedance across apertures when traversed by individual particles, e.g. by using the Coulter principle
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 2015/0053 in liquids, e.g. trouble
• CPC Class Codes: G01N 2015/1019 Associating Coulter-counter and optical flow cytometer [OFC]
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]
• CPC Class Codes: Y10T 436/106664 Blood serum or blood plasma standard or control
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/117497 with a continuously flowing sample or carrier stream

Abstract

A reagent for analyzing solid components in urine comprising: (i) a buffer agent for maintaining pH at 5.0 to 9.0, (ii) an osmotic pressure compensating agent for maintaining osmotic pressure at 100 mOsm/kg to 600 mOsm/kg, (iii) a first dye which is a condensed benzene derivative, (iv) a second fluorescent dye capable of staining a damaged cell, and (v) a chelating agent.

Claims

1. A method capable of differentiating at least crystalline components, erythrocytes, yeast-like fungi and leukocytes in urine which comprises the steps of:

mixing the urine with a reagent for analyzing solid components in urine comprising;

(i) a buffer agent for maintaining pH at 5.0 to 9.0,

(ii) an osmotic pressure compensating agent for maintaining osmotic pressure at 100 mOsm/kg to 600 mOsm/kg,

(iii) a first dye which is a condensed benzene derivative selected from the group consisting of the following formula;

3,3'"'"'-(di-n-hexyl)-2,2'"'"'-oxacarbocyanine iodide (NK-2280, DiOC6(3)) ##STR14## 2-(p-dimethylaminostyryl)-3-methylbenzooxazolium iodide (NK-528) ##STR15## 2-(p-dimethylaminostyryl)-1,3,3-trimethyl-3H-indolium iodide (NK-97) ##STR16## and is capable of differentiating crystalline components, erythrocytes and yeast-like fungi,

(iv) a second fluorescent dye capable of staining a damaged cell, and

(v) a chelating agent

to stain the desired solid components in the urine, applying an excitation light to solid components in the above stained urine, and

measuring the scattered light and the fluorescent light emitted from the solid components.

2. A method according to claim 1, wherein an electric resistance signal is measured after the solid components in the urine are stained.

3. A method according to claim 1, wherein the reagent for analyzing is mixed with the urine after the reagent is previously warmed.

4. A method according to claim 1, wherein said buffer agent maintains pH at 6.5 to 7.5.

5. A method according to claim 4, wherein said buffer agent maintains pH at 6.8 to 7.2.

6. A method according to claim 1, wherein said osmotic pressure compensating agent maintains osmotic pressure at 150 mOsm/kg to 500 mOsm/kg.

7. A method according to claim 1, wherein the electrical conductivity of the reagent is adjusted to 1 to 10 mS/cm.

8. A method according to claim 7, wherein the electrical conductivity of the reagent is adjusted to 4 to 7 mS/cm.

9. A method according to claim 1, wherein the reagent for analyzing is previously warmed to 30°

C. to 40°

C.

10. A method according to claim 9, wherein the reagent for analyzing is previously warmed to 33°

C. to 37°

C.

14. A method according to claim 2, wherein said buffer agent maintains pH at 6.5 to 7.5.

15. A method according to claim 14, wherein said buffer agent maintains pH at 6.8 to 7.2.

11. A method capable of differentiating at least crystalline components, erythrocytes, yeast-like fungi and leukocytes in urine which comprises the steps of:

mixing the urine with a reagent for analyzing solid components in urine comprising;

(I) a buffer agent for maintaining pH at 5.0 to 9.0,

(II) an osmotic pressure compensating agent for maintaining osmotic pressure at 100 mOsm/kg to 600 mOsm/kg,

(III) a dye capable of being excited by a red wave-length light, which is selected from the group consisting of the following;

##STR17## Rhodnile Blue, Capri Blue BB, Basic Blue 124 and Basic Blue 1, and is capable of differentiating crystalline components, erythrocytes and yeast-like fungi, and

(IV) a chelating agent

to stain the desired solid components in the urine,

applying an excitation light to solid components in the above stained urine, and

measuring the scattered light and the fluorescent light emitted from the solid components.

12. A method according to claim 11, wherein an electric conductivity signal is measured after the solid components in the urine are stained.

13. A method according to claim 11, wherein the reagent for analyzing is mixed with the urine after the reagent is previously warmed.

16. A method according to claim 11, wherein said osmotic pressure compensating agent maintains osmotic pressure at 150 mOsm/kg to 500 mOsm/kg.

17. A method according to claim 11, wherein the electrical conductivity of the reagent is adjusted to 1 to 10 mS/cm.

18. A method according to claim 17, wherein the electrical conductivity of the reagent is adjusted to 4 to 7 mS/cm.

19. A method according to claim 11, wherein the reagent for analyzing is previously warmed to 30°

C. to 40°

C.

20. A method according to claim 19, wherein the reagent for analyzing is previously warmed to 33°

C. to 37°

C.