CN107091934A - A kind of direct bilirubin detecting kit and collocation method and its utilization - Google Patents
A kind of direct bilirubin detecting kit and collocation method and its utilization Download PDFInfo
- Publication number
- CN107091934A CN107091934A CN201610089227.7A CN201610089227A CN107091934A CN 107091934 A CN107091934 A CN 107091934A CN 201610089227 A CN201610089227 A CN 201610089227A CN 107091934 A CN107091934 A CN 107091934A
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- emulgen
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- direct bilirubin
- detecting kit
- bilirubin detecting
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- 238000008789 Direct Bilirubin Methods 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 17
- 206010023138 Jaundice neonatal Diseases 0.000 claims abstract description 18
- 239000003153 chemical reaction reagent Substances 0.000 claims description 36
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 24
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 8
- 238000007792 addition Methods 0.000 claims description 5
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 4
- OVBJJZOQPCKUOR-UHFFFAOYSA-L EDTA disodium salt dihydrate Chemical compound O.O.[Na+].[Na+].[O-]C(=O)C[NH+](CC([O-])=O)CC[NH+](CC([O-])=O)CC([O-])=O OVBJJZOQPCKUOR-UHFFFAOYSA-L 0.000 claims description 4
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 239000008213 purified water Substances 0.000 claims description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 4
- CMZUMMUJMWNLFH-UHFFFAOYSA-N sodium metavanadate Chemical compound [Na+].[O-][V](=O)=O CMZUMMUJMWNLFH-UHFFFAOYSA-N 0.000 claims description 4
- 150000004683 dihydrates Chemical class 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 239000001509 sodium citrate Substances 0.000 claims description 2
- 206010023126 Jaundice Diseases 0.000 abstract description 11
- 238000001514 detection method Methods 0.000 abstract description 8
- 238000005259 measurement Methods 0.000 abstract description 2
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 26
- 239000008280 blood Substances 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 238000012360 testing method Methods 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- WQEVDHBJGNOKKO-UHFFFAOYSA-K vanadic acid Chemical group O[V](O)(O)=O WQEVDHBJGNOKKO-UHFFFAOYSA-K 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 2
- 230000008676 import Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000002277 temperature effect Effects 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
- G01N33/728—Bilirubin; including biliverdin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of direct bilirubin detecting kit and collocation method and its utilization, the problem of jaundice has interference to direct bilirubin detecting in neonate's sample is solved.The present invention includes direct bilirubin detecting kit body, and Emulgen A-90 and/or Emulgen B-66.The present invention is prevented effectively from the interference that icterus neonatorum is brought to Samples detection, and it is the false higher phenomenon of measurement result that sample is brought to solve icterus neonatorum, greatly improves the accuracy in detection of icterus neonatorum sample.
Description
Technical field
The present invention relates to a kind of kit, and in particular to a kind of direct bilirubin detecting kit, and there is provided the reagent
The collocation method of box and utilization.
Background technology
The application of bilirubin direct clinically is relatively broad, has higher clinical value to the judgement of disease, to new life
Youngster's sample, which is determined, also important clinical meaning.Non-neonate, can be direct because internal bilirubin metabolism produces jaundice extremely
Bilirubin direct level in blood is caused to raise.That is, bilirubin direct level rise can both be caused by physiologic jaundice, can also
Caused by pathological jaundice.
The common method of bilirubin direct is Vanadic acid oxidation, and existing Vanadic acid oxidation determines icterus neonatorum mark
This when, detection data are easily disturbed by jaundice, increase its exception, very big interference is brought to clinical diagnosis, causes clinical assays
As a result inaccurate, influence clinician correctly judges patient status.And after clinic is accurately judged as physiologic jaundice, it is surveyed
Definite value still can be extremely higher, thus cause clinically can not accurate quantitative analysis bilirubin direct actual concentration.
The content of the invention
The technical problems to be solved by the invention are icterus neonatorum samples have to the measure of existing bilirubin direct it is dry
Disturb, it is therefore intended that a kind of direct bilirubin detecting kit of offer, jaundice is to direct bilirubin detecting in solution neonate's sample
The problem of interference.
The present invention is achieved through the following technical solutions：
A kind of direct bilirubin detecting kit, including direct bilirubin detecting kit body, and Emulgen A-90 and/
Or Emulgen B-66.
The present invention is effectively reduced or even eliminated by adding Emulgen A-90 and/or Emulgen B-66 in reagent
The interference that icterus neonatorum sample is brought, improves the authenticity of direct bilirubin detecting.
Learnt by experiment, while when adding Emulgen A-90 and Emulgen two kinds of materials of B-66, icterus neonatorum
The interference that jaundice is brought in sample is minimum, detects the accuracy highest of obtained data；And the Emulgen A-90's is dense
Spend for 1~10g/L, when the concentration of the Emulgen B-66 is 1~10g/L, as shown in table 2, can effectively eliminate neonate yellow
The interference that subcutaneous ulcer sample is brought, makes Detection results most accurate.
Preferably, the direct bilirubin detecting kit body includes reagent 1 and reagent 2,
The reagent 1 includes：
100~150mmol/L of citric acid
1.5~3.5g/L of NaOH
4.16~5.12g/L of HAS
1.0~1.5ml/L of PC300；
The reagent 2 includes：
20~50mmol/L of citrate dihydrate trisodium
2.2~4.2g/L of citric acid
0.8~1.2g/L of sodium metavanadate
0.5~1.0ml/L of Sodium azide
0.5~3.0mmol/L of disodium ethylene diamine tetraacetate.
A kind of collocation method of above-mentioned direct bilirubin detecting kit, including：
（1）The configuration of reagent in direct bilirubin detecting kit body is completed first,
（2）After Emulgen A-90 and/or Emulgen B-66 are dissolved with water respectively, it is slow added into reagent, when
When Emulgen A-90 and Emulgen B-66 are added, first add Emulgen A-90 and be sufficiently stirred for mixing, it is then slow again
Emulgen B-66 are added to be sufficiently stirred for mixing；
（3）Constant volume.
Emulgen A-90 and dissolving of the two kinds of materials of Emulgen B-66 in direct bilirubin detecting kit body
Degree is relatively low, and being directly added into bilirubin direct reagent to be completely dissolved, and the effect played in reagent can be caused to subtract completely
It is weak.The present invention is tried using direct bilirubin detecting is then added to after dissolving Emulgen A-90 and Emulgen B-66 in water
In agent box body, the problem of it can not be completely dissolved is prevented effectively from, meanwhile, after being dissolved and added by the method for the present invention
The declines that conventional addition manner causes it played in reagent can be prevented effectively from, effect is notable.
Present inventor draws after experiment and research many times, as the Emulgen A-90 and Emulgen
When B-66 solution temperature is 35~39 DEG C, the function influence played to it in reagent is minimum, and can have at such a temperature
Effect is completely dissolved Emulgen A-90 and Emulgen B-66, plays it and preferably acts on.
Preferably, the solution temperature of the Emulgen A-90 and/or Emulgen B-66 are 37 DEG C.
Preferably, the direct bilirubin detecting kit body includes reagent 1 and reagent 2,
The configuration process of the reagent 1 is as follows：
Sequentially citric acid, sodium hydroxide, HAS, PC300 are added into purified water, after the completion of the dissolving of former material
Latter material, water suction piping and druming, then Emulgen A-90 and/or Emulgen B-66 are added repeatedly after PC300 additions are added again
Enter rear constant volume；
The configuration process of the reagent 2 is as follows：
Sequentially Sodium Citrate, usp, Dihydrate Powder, citric acid, sodium metavanadate, Sodium azide, disodium ethylene diamine tetraacetate are added into purified water,
Latter material, last constant volume are added again after the completion of the dissolving of former material.
A kind of direct bilirubin detecting kit of the present invention, its bilirubin direct for being applied to all blood samples is surveyed
It is fixed, but its utilization best results on icterus neonatorum sample, it can be prevented effectively from what jaundice in icterus neonatorum sample was brought
Interference, the degree of accuracy highest for detecting it.
The present invention compared with prior art, has the following advantages and advantages：
1st, the present invention can the interference that is brought by Samples detection of effectively anti-icterus neonatorum, by Emulgen A-90 and
Emulgen B-66 can significantly solve the false higher phenomenon of measurement result that icterus neonatorum is brought very much, make icterus neonatorum mark
This accuracy in detection is greatly improved；
2nd, collocation method of the invention can effectively be such that Emulgen A-90 and Emulgen B-66 is sufficiently dissolved in reagent, send out
Emulgen A-90 and Emulgen B-66 are waved preferably to act on.
Embodiment
For the object, technical solutions and advantages of the present invention are more clearly understood, with reference to embodiment, to present invention work
Further to describe in detail, exemplary embodiment and its explanation of the invention is only used for explaining the present invention, is not intended as to this
The restriction of invention.
Embodiment 1
As shown in figure 1, a kind of direct bilirubin detecting kit of the invention, concrete composition composition is as shown in table 1.
Table 1
Collect birth 28 days within newborn blood sample, screening determination of bilirubin concentration 34.2umol/L~
205.23umol/L（The index of physiological jaundice of newborn）Between 40 samples, to the examination using embodiment in above-mentioned table 1
Agent detects that testing result is as shown in table 2 to 40 samples.
Table 2
Standard value in above-mentioned table 2 is using the bilirubin direct reagent not disturbed by icterus neonatorum of import test
The value arrived, the concrete model of the bilirubin direct reagent of the import is the bilirubin direct of Japanese Wako Pure Chemical Industries, Ltd.
Determine kit（Vanadic acid oxidation）.
The present invention has inquired into direct bilirubin detecting kit body simultaneously, and with TX-100, Tween-20, Brij-
35th, dodecyl sodium sulfate, Tween-80, the reagent of NP40, NP1055 TX-405 isoreactivity agent are in icterus neonatorum sample
The situation of jaundice interference, specific test result is as shown in table 3.
Table 3
By above-mentioned table 2 and table 3, while Emulgen A-90 and Emulgen B-66 are added into direct bilirubin detecting
In reagent box body, i.e., when being detected using the direct bilirubin detecting kit of the present invention, it can effectively mitigate or even eliminate
Interference of the physiological jaundice of newborn to it in direct bilirubin detecting, makes detection data more accurate.
The present embodiment also inquired into using existing direct bilirubin detecting kit detected when, neonate's physiological
Influence of the jaundice to direct bilirubin detecting result.Direct bilirubin detecting kit body is newly good for using Sichuan in the present embodiment
Direct bilirubin detecting kit of the health into Biological Co., Ltd..In use, according to the method for the present invention by Emulgen
A-90 and Emulgen B-66 are added in reagent, and Emulgen A-90 concentration is 4.5g/L, Emulgen B-66 concentration
For 5.5g/L.
Using the former direct bilirubin detecting reagent of above-mentioned Sichuan Xinjiankangcheng Biolog Technology Co., Ltd., addition
Direct bilirubin detecting reagent after Emulgen A-90 and Emulgen B-66 detects to neonate's blood sample, testing result
As shown in table 4.
Table 4
Can further it be proved by the data in above-mentioned table 4：Emulgen A-90 and Emulgen B-66 are directly appended to
In direct bilirubin detecting reagent, when being detected to neonate's blood sample, physiological jaundice of newborn can be effectively excluded to detection
As a result interference, effect is notable.
Above-described embodiment, has been carried out further to the purpose of the present invention, technical scheme and beneficial effect
Describe in detail, should be understood that the embodiment that the foregoing is only the present invention, be not intended to limit the present invention
Protection domain, within the spirit and principles of the invention, any modification, equivalent substitution and improvements done etc. all should be included
Within protection scope of the present invention.
Claims (8)
1. including direct bilirubin detecting kit body, and Emulgen A-90 and/or Emulgen B-66.
2. a kind of direct bilirubin detecting kit according to claim 1, it is characterised in that the Emulgen A-90
Concentration be 1~10g/L, the concentration of the Emulgen B-66 is 1~10g/L.
3. a kind of direct bilirubin detecting kit according to claim 1 or 2, it is characterised in that the direct courage is red
Element, which determines reagent box body, includes reagent 1 and reagent 2,
The reagent 1 includes：
100~150mmol/L of citric acid
1.5~3.5g/L of NaOH
4.16~5.12g/L of HAS
1.0~1.5ml/L of PC300；
The reagent 2 includes：
20~50mmol/L of citrate dihydrate trisodium
2.2~4.2g/L of citric acid
0.8~1.2g/L of sodium metavanadate
0.5~1.0ml/L of Sodium azide
0.5~3.0mmol/L of disodium ethylene diamine tetraacetate.
4. a kind of collocation method of direct bilirubin detecting kit as described in any one of claims 1 to 3, its feature exists
In, including：
（1）The configuration of reagent in direct bilirubin detecting kit is completed first,
（2）After Emulgen A-90 and/or Emulgen B-66 are dissolved with water respectively, it is slow added into reagent, when
When Emulgen A-90 and Emulgen B-66 are added, first add Emulgen A-90 and be sufficiently stirred for mixing, it is then slow again
Emulgen B-66 are added to be sufficiently stirred for mixing；
（3）Constant volume.
5. a kind of collocation method of direct bilirubin detecting kit as claimed in claim 4, it is characterised in that described
Emulgen A-90 and/or Emulgen B-66 solution temperature are 35~39 DEG C.
6. a kind of collocation method of direct bilirubin detecting kit as claimed in claim 5, it is characterised in that described
Emulgen A-90 and/or Emulgen B-66 solution temperature are 37 DEG C.
7. a kind of collocation method of direct bilirubin detecting kit as described in any one of claim 4~6, its feature exists
In, the direct bilirubin detecting kit body includes reagent 1 and reagent 2,
The configuration process of the reagent 1 is as follows：
Sequentially citric acid, sodium hydroxide, HAS, PC300 are added into purified water, completed when former material dissolves
Add latter material again afterwards, water suction piping and druming repeatedly after PC300 additions, then by Emulgen A-90 and/or Emulgen B-66
Constant volume after addition；
The configuration process of the reagent 2 is as follows：
Sequentially Sodium Citrate, usp, Dihydrate Powder, citric acid, sodium metavanadate, Sodium azide, disodium ethylene diamine tetraacetate are added into purified water,
Latter material, last constant volume are added again after the completion of the dissolving of former material.
8. a kind of direct bilirubin detecting kit as described in any one of claims 1 to 3 is on icterus neonatorum sample
With.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610089227.7A CN107091934B (en) | 2016-02-18 | 2016-02-18 | A kind of direct bilirubin detecting kit and configuration method and its utilization |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610089227.7A CN107091934B (en) | 2016-02-18 | 2016-02-18 | A kind of direct bilirubin detecting kit and configuration method and its utilization |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107091934A true CN107091934A (en) | 2017-08-25 |
| CN107091934B CN107091934B (en) | 2018-07-06 |
Family
ID=59648646
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610089227.7A Active CN107091934B (en) | 2016-02-18 | 2016-02-18 | A kind of direct bilirubin detecting kit and configuration method and its utilization |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109813918A (en) * | 2019-01-11 | 2019-05-28 | 河北省医疗器械与药品包装材料检验研究院(河北省医疗器械技术审评中心) | A kind of total bilirubin determination reagent kit |
| CN111455018A (en) * | 2020-03-03 | 2020-07-28 | 天津大学 | Direct bilirubin detection kit containing bacillus subtilis laccase |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0484133A2 (en) * | 1990-10-30 | 1992-05-06 | Wako Pure Chemical Industries Ltd | Method for measuring bilirubin |
| US20050095658A1 (en) * | 2002-01-30 | 2005-05-05 | Denka Seiken Co Ltd | Method of quantifying cholesterol in high density lipoprotein and reagent compositions |
| EP2105739A1 (en) * | 2008-03-26 | 2009-09-30 | Fujifilm Corporation | Dry analytical element capable of reducing influence of hemolysis for body fluid component measurement |
| CN104155438A (en) * | 2014-09-09 | 2014-11-19 | 湖北科技学院 | Determination reagent of total bile acid |
| CN104406971A (en) * | 2014-11-28 | 2015-03-11 | 山东博科生物产业有限公司 | Direct bilirubin detection reagent |
-
2016
- 2016-02-18 CN CN201610089227.7A patent/CN107091934B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0484133A2 (en) * | 1990-10-30 | 1992-05-06 | Wako Pure Chemical Industries Ltd | Method for measuring bilirubin |
| US20050095658A1 (en) * | 2002-01-30 | 2005-05-05 | Denka Seiken Co Ltd | Method of quantifying cholesterol in high density lipoprotein and reagent compositions |
| EP2105739A1 (en) * | 2008-03-26 | 2009-09-30 | Fujifilm Corporation | Dry analytical element capable of reducing influence of hemolysis for body fluid component measurement |
| CN104155438A (en) * | 2014-09-09 | 2014-11-19 | 湖北科技学院 | Determination reagent of total bile acid |
| CN104406971A (en) * | 2014-11-28 | 2015-03-11 | 山东博科生物产业有限公司 | Direct bilirubin detection reagent |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109813918A (en) * | 2019-01-11 | 2019-05-28 | 河北省医疗器械与药品包装材料检验研究院(河北省医疗器械技术审评中心) | A kind of total bilirubin determination reagent kit |
| CN109813918B (en) * | 2019-01-11 | 2021-04-09 | 河北省药品医疗器械检验研究院 | Total bilirubin determination kit |
| CN111455018A (en) * | 2020-03-03 | 2020-07-28 | 天津大学 | Direct bilirubin detection kit containing bacillus subtilis laccase |
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| Publication number | Publication date |
|---|---|
| CN107091934B (en) | 2018-07-06 |
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