CN103439484B - A kind of Color-development quality control material and application thereof - Google Patents
A kind of Color-development quality control material and application thereof Download PDFInfo
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Abstract
The invention discloses a kind of Color-development quality control material and application thereof, belong to Quality Control thing technical field.Described Color-development quality control material is that in common Quality Control thing, add concentration be that the coloring agent of 0.001g/ml ~ 1g/ml obtains.This Color-development quality control material can naked eyes with detect sample harmonizing yinyang reference substance and distinguish, when being convenient to test, Quality Control thing is randomly dispersed in and detects in sample, the better monitoring effect of performance Quality Control thing, and avoids adding Wrong, missing and add Quality Control thing.The more important thing is that this Color-development quality control material well can keep its original stable Biochemistry laboratory course, complete enzyme linked immunological experiment accurately.
Description
Technical field
The invention belongs to Quality Control thing technical field, be specifically related to a kind of Color-development quality control material and application thereof.
Background technology
Enzyme-linked immunosorbent assay (ELISA) is detection method the most frequently used in clinical examination, because of its sensitivity and specificity is higher, robotization, the advantage such as cheap, therefore very advantageous in high-throughout blood testing is the irreplaceable method that current Blood Transfusion Services of China carries out blood safety detection.Third liver, syphilis, hepatitis B and acquired immune deficiency syndrome (AIDS) are four infectious disease indexs that the blood of all collections of Ministry of Public Health's regulation must detect, and its detection method is ELISA.
When doing ELISA test, a hole or porous Internal Quality Control thing must be done to monitor the validity of each test findings together with the blood sample detected.The matrix of Internal Quality Control thing comes from the serum of people, and therefore, its color of Quality Control thing that current most of testing agency uses is the same with the serum specimen of detection, is all water white transparency or micro-yellow (water white transparency is serum color after treatment).When mass detection blood sample, full-automatic loading device is adopted to be added in the ELISA Plate hole of 300ul by 50 μ l Quality Control things.Because the blood preparation of its color and detection is difficult to differentiate, can only fixed hole position in test, what tester just can know which hole does is Quality Control thing, and the requirement in fact doing Quality Control thing must be randomly distributed in the middle of blood preparation, test the monitoring effect just playing Quality Control thing together with detection sample.And the color of the negative control of current most of reagent manufacturer production, positive control and Quality Control thing is quite similar, naked eyes almost can not distinguish.Therefore, whether experimenter is also difficult to distinguish yin and yang attribute tester and Quality Control thing has and adds wrong position, or the phenomenon mixed.The test findings of Quality Control thing is directly connected to the validity of experimental data by the gross.In real work because of Quality Control thing color bad with yin and yang attribute tester and the serum specimen of detection distinguish and occur adding Wrong, missing and to add or the phenomenon of quantity not sufficient happens occasionally; the result of Quality Control thing is related to the validity of test findings by the gross; test findings is invalid usually can cause test to reform, and brings unnecessary trouble and waste to patient and staff.Hepatitis C, microspironema pallidum, hepatitis B surface antigen and AIDS virus are four infectious disease indexs that the blood of all collections of Ministry of Public Health's regulation must detect, test is caused to be reformed because Quality Control thing result is nonconforming, patient may be had influence on time serious transfuse blood in time, jeopardize the life of patient.Therefore, the improvement for the color trait of Quality Control thing becomes particularly important.
Summary of the invention
The object of the invention is to solve the problem and provide one and can keep the original Biochemistry laboratory course of Quality Control thing, and the Color-development quality control material that can with the naked eye distinguish and application thereof.
The technical solution adopted in the present invention is:
A kind of Color-development quality control material, described Color-development quality control material is that in common Quality Control thing, add concentration be that the coloring agent of 0.001g/ml ~ 1g/ml obtains.
Further, in described Color-development quality control material, original Biochemistry laboratory course of common Quality Control thing can not change.
Preferably, described coloring agent is chemosynthesis pigment or natural colouring matter.
More preferably, described chemosynthesis pigment is famille rose, lemon yellow or sunset yellow.
More preferably, described natural colouring matter is beet red.
Preferably, described common Quality Control thing is AIDS virus Quality Control thing, the third liver Quality Control thing, syphilis Quality Control thing or hepatitis B Quality Control thing.
Further, when described common Quality Control thing is AIDS virus Quality Control thing, syphilis Quality Control thing or hepatitis B Quality Control thing, the concentration adding coloring agent is 0.1g/ml.
Further, when described common Quality Control thing is the third liver Quality Control thing, the concentration adding coloring agent is 0.2g/ml.
Further, described Color-development quality control material can ensure the stability of enzyme-linked immunosorbent assay.
The present invention has the following advantages:
The color trait that the present invention overcomes existing Quality Control thing is not enough, a kind of Color-development quality control material is provided, this display Quality Control thing can ensure that common Quality Control thing can not only keep its Biochemistry laboratory course originally, the process of monitoring enzyme linked immunoassay, and naked eyes can be easily passed through the blood sample of itself and positive control, negative control thing and detection is made a distinction, and be convenient to Quality Control thing to be arranged at random in the blood sample of detection, make whole experiment process more safe and reliable, the value of Quality Control thing more can accurately reaction test result.
Accompanying drawing explanation
Fig. 1 is that the hepatitis B Quality Control thing that develops the color in embodiment 1 is applied to the colour developing result figure of enzyme-linked immunosorbent assay;
Fig. 2 is that the syphilis Quality Control thing that develops the color in embodiment 2 is applied to the colour developing result figure of enzyme-linked immunosorbent assay;
Fig. 3 is that the AIDS virus Quality Control thing that develops the color in embodiment 3 is applied to the colour developing result figure of enzyme-linked immunosorbent assay;
Fig. 4 is that the third liver Quality Control thing that develops the color in embodiment 4 is applied to the colour developing result figure of enzyme-linked immunosorbent assay.
Embodiment
Below in conjunction with drawings and embodiments, the present invention is further detailed explanation.
Embodiment 1
Present embodiments provide a kind of preparation method of the hepatitis B Quality Control thing that develops the color, comprise the steps:
(1) 1.0g dyestuff lemon yellow is taken with analytical balance, with 10ml deionized water dissolving, dilution.Centrifugal 1 minute of the rotating speed that hydro-extractor is 1000 revs/min, stand-by.
(2) Anti-HBs antibody Ag Quality Control thing 1 (specification: 0.5ml/ props up or 1ml/ props up) is taken out, equilibrium at room temperature 30 minutes.
(3) draw the lemon yellow dyestuff that dissolved of 1ul to add 0.5ml/ and to prop up or 1ml/ props up in anti-HBsAg Quality Control thing, cover lid, puts upside down mixing 1 minute back and forth, the very fast level dyeing of visible Quality Control thing.
(4) preservation of Quality Control thing: under being stored in-20 DEG C of conditions.
(5) use before be put in thaw one day in 4 DEG C of refrigerators after to be put in equilibrate at room temperature more for subsequent use for half an hour.
This colour developing hepatitis B Quality Control thing is applied to ELISA experiment, and its experimental procedure is as follows:
(1) to make even the Anti-HBs antibody Ag ELISA reagent weighed, Article 1, in enzyme mark hole, add negative control 2 hole (A1, B1), positive control 2 hole (C1, D1) Quality Control thing 1 hole (E1), the each 100ul in every hole, in Article 2 and Article 3 and Article 4 microplate, (A2-H2, A3-H3, A4-H4) adds each 100ul in the every hole of Quality Control thing not adding coloring agent in totally 24 holes, (A, B, C, D, E, F, G, H and A5-H5, A6-H6, A7-H7) hole of Article 5, six and seven adds colour developing hepatitis B Quality Control thing, each 100ul in every hole in totally 24 holes.Its colour developing result is with reference to Fig. 1.
(2) microplate is placed in 37 DEG C of incubators and hatches 1 hour.
(3) washing lotion configured washes plate 5 times, and button is dry.
(4) every enzyme-added 100ul in hole, continues at 37 DEG C of incubators and hatches 1 hour.
(5) washing lotion configured washes plate 5 times, adds substrate (each 50ul of A, B liquid).
(6) 37 DEG C of incubators hatch 30 minutes.
(7) every hole adds stop buffer 100ul, and microplate develops the color, and detects OD value by microplate reader.
The OD value that the OD value measured by protoplasm control quality testing and colour developing hepatitis B Quality Control quality testing measure is carried out t and is checked and compare, P=0.51 > 0.05, two groups of data are without obvious significant difference, illustrate that the Quality Control thing after this colouring method process can well maintain the Biochemistry laboratory course of Quality Control thing, the enzyme linked immunological that is applied to that can be stable is tested.For confirming that the Quality Control thing after improving does not produce any impact to Blood Transfusion Services's routine work, detect in Blood Transfusion Services the microplate of hepatitis B add improvement in the blank well reserved after Quality Control thing 100ul, enter in full-automatic FAME aftertreatment instrument with microplate and test one month, doing microplate number is altogether 211 plates, by improve after Quality Control thing institute's value and protoplasm control thing numeric ratio comparatively, (P=0.65>0.05, difference does not have statistical significance).Quality Control thing after improving is joined any hole in microplate at random together with detection sample, do microplate 30 pieces altogether, the value improving rear Quality Control thing gained is compared with protoplasm control thing OD value, (P=0.43>0.05, difference does not have statistical significance).
Embodiment 2
Present embodiments provide a kind of preparation method of the syphilis Quality Control thing that develops the color, comprise the steps:
(1) 1.0g dyestuff lemon yellow is taken with analytical balance, with 10ml deionized water dissolving, dilution.Centrifugal 1 minute of the rotating speed that hydro-extractor is 1000 revs/min, stand-by.
(2) syphilis Quality Control thing 1 (specification: 0.5ml/ props up or 1ml/ props up) is taken out, equilibrium at room temperature 30 minutes.
(3) draw the lemon yellow dyestuff that dissolved of 1ul to add 0.5ml/ and to prop up or 1ml/ props up in syphilis Quality Control thing, cover lid, puts upside down mixing 1 minute back and forth, the very fast level dyeing of visible Quality Control thing.
(4) preservation of Quality Control thing: under being stored in-20 DEG C of conditions.
(5) use before be put in thaw one day in 4 DEG C of refrigerators after to be put in equilibrate at room temperature more for subsequent use for half an hour.
This colour developing syphilis Quality Control thing is applied to ELISA experiment, and its experimental procedure is as follows:
(1) to make even the syphilis ELISA reagent weighed, Article 1, in enzyme mark hole, add negative control 2 hole (A1, B1), positive control 2 hole (C1, D1) Quality Control thing 1 hole (E1), the each 100ul in every hole, Article 2 and the interior (A2-H2 of Article 3 microplate, A3-H3) totally 16 holes add each 100ul in the every hole of Quality Control thing not adding coloring agent, add the colour developing syphilis Quality Control thing each 100ul in every hole in (A, B, C, D, E) hole of Article 5, six and seven.Its colour developing result is with reference to Fig. 2.
(2) microplate is placed in 37 DEG C of incubators and hatches 1 hour.
(3) washing lotion configured washes plate 5 times, and button is dry.
(4) every enzyme-added 100ul in hole, continues at 37 DEG C of incubators and hatches 1 hour.
(5) washing lotion configured washes plate 5 times, adds substrate (each 50ul of A, B liquid).
(6) 37 DEG C of incubators hatch 30 minutes.
(7) every hole adds stop buffer 50ul, and microplate develops the color, and detects OD value by microplate reader.
(8) the OD value that the OD value measured by protoplasm control quality testing and colour developing syphilis Quality Control quality testing measure is carried out t and is checked and compare, P=0.88 > 0.05, two groups of data are without obvious significant difference, illustrate that the Quality Control thing after this colouring method process can well maintain the Biochemistry laboratory course of Quality Control thing, the enzyme linked immunological that is applied to that can be stable is tested.For confirming that the Quality Control thing after improving does not produce any impact to Blood Transfusion Services's routine work, detect in Blood Transfusion Services the microplate of syphilis add improvement in the blank well reserved after Quality Control thing 100ul, enter in full-automatic FAME aftertreatment instrument with microplate and test one month, doing microplate number is altogether 200 plates, by improve after Quality Control thing institute's value and protoplasm control thing numeric ratio comparatively, (P=0.72>0.05, difference does not have statistical significance).Quality Control thing after improving is joined any hole in microplate at random together with detection sample, do microplate 30 pieces altogether, the value improving rear Quality Control thing gained is compared with protoplasm control thing OD value, (P=0.55>0.05, difference does not have statistical significance).
Embodiment 3
Present embodiments provide a kind of preparation method of the AIDS virus Quality Control thing that develops the color, comprise the steps:
(1) 1.0g dyestuff sunset yellow is taken with analytical balance, with 10ml deionized water dissolving, dilution.Centrifugal 1 minute of the rotating speed that hydro-extractor is 1000 revs/min, stand-by.
(2) AIDS virus Quality Control thing 1 (specification: 0.5ml/ props up or 1ml/ props up) is taken out, equilibrium at room temperature 30 minutes.
(3) draw the sunset yellow dyestuff that dissolved of 1ul to add 0.5ml/ and to prop up or 1ml/ props up in hepatitis B Quality Control thing, cover lid, puts upside down mixing 1 minute back and forth, the very fast level dyeing of visible Quality Control thing.
(4) preservation of Quality Control thing: under being stored in-20 DEG C of conditions.
(5) use before be put in thaw one day in 4 DEG C of refrigerators after to be put in equilibrate at room temperature more for subsequent use for half an hour.
This colour developing AIDS virus Quality Control thing is applied to ELISA experiment, and its experimental procedure is as follows:
(1) to make even the AIDS virus ELISA reagent weighed, Article 1, in enzyme mark hole, add negative control 2 hole (A1, B1), positive control 2 hole (C1, D1) Quality Control thing 1 hole (E1), the each 100ul in every hole, Article 2 and the interior (A2-H2 of Article 3 microplate, A3-H3) totally 24 holes add each 50ul in the every hole of Quality Control thing not adding coloring agent, and (A, B, C, D, E, F, G, H and A5-H5, A6-H6, A7-H7) hole of Article 5, six and seven adds the colour developing AIDS virus Quality Control thing each 100ul in every hole in totally 24 holes.Its colour developing result is with reference to Fig. 3.
(2) microplate is placed in 37 DEG C of incubators and hatches 1 hour.
(3) washing lotion configured washes plate 5 times, and button is dry.
(4) every enzyme-added 100ul in hole, continues at 37 DEG C of incubators and hatches 1 hour.
(5) washing lotion configured washes plate 5 times, adds substrate (each 50ul of A, B liquid).
(6) 37 DEG C of incubators hatch 30 minutes.
(7) every hole adds stop buffer 100ul, and microplate develops the color, and detects OD value by microplate reader.
(8) the OD value that the OD value measured by protoplasm control quality testing and colour developing AIDS virus Quality Control thing measure is carried out t and is checked and compare, P=0.45 > 0.05, two groups of data are without obvious significant difference, illustrate that the Quality Control thing after this colouring method process can well maintain the Biochemistry laboratory course of Quality Control thing, the enzyme linked immunological that is applied to that can be stable is tested.For confirming that the Quality Control thing after improving does not produce any impact to Blood Transfusion Services's routine work, detect in Blood Transfusion Services the microplate of AIDS virus add improvement in the blank well reserved after Quality Control thing 100ul, enter in full-automatic FAME aftertreatment instrument with microplate and test one month, doing microplate number is altogether 163 plates, by improve after Quality Control thing institute's value and protoplasm control thing numeric ratio comparatively, (P=0.77>0.05, difference does not have statistical significance).Quality Control thing after improving is joined any hole in microplate at random together with detection sample, do microplate 20 pieces altogether, the value improving rear Quality Control thing gained is compared with protoplasm control thing OD value, (P=0.41>0.05, difference does not have statistical significance).
Embodiment 4
Present embodiments provide the preparation method of a kind of colour developing the third liver Quality Control thing, comprise the steps:
(1) 1.0g dyestuff is taken with analytical balance carmine, with 5ml deionized water dissolving, dilution.Centrifugal 1 minute of the rotating speed that hydro-extractor is 1000 revs/min, stand-by.
(2) HCV Quality Control thing (specification: 0.5ml/ props up or 1ml/ props up) is taken out, equilibrium at room temperature 30 minutes.
(3) cochineal dye that absorption 1ul has dissolved adds in 1.0mlHCV Quality Control thing, and cover lid, puts upside down mixing 1 minute back and forth, the very fast level dyeing of visible Quality Control thing.
(4) preservation of Quality Control thing: under being stored in-20 DEG C of conditions.
(5) use before be put in thaw one day in 4 DEG C of refrigerators after to be put in equilibrate at room temperature more for subsequent use for half an hour.
The third liver Quality Control thing that this developed the color is applied to ELISA experiment, and its experimental procedure is as follows:
(1) to make even the HCV ELISA reagent weighed, every hole adds 100ul dilution, Article 1, in reagent, add negative control 2 hole (A1, B1), positive control 2 hole (C1, D1) Quality Control thing 1 hole (E1), the each 10ul in every hole, Article 2 and the interior (A2-H2 of Article 3 microplate, A3-H3,) totally 16 holes add each 10ul in the every hole of Quality Control thing not adding coloring agent, (A, B, C, D, E, F, G, H and A7-H7, A8-H8, A9-H9) Nei Gong24 hole, hole of the 7th article, 8 articles and 9 articles adds each 10ul in the every hole of Quality Control thing through famille rose dyeing.Its colour developing result is with reference to Fig. 4 (for only adding dilution in A4-H4, A5-H5, A6-H6 hole).
(2) microplate is placed in 37 DEG C of incubators and hatches 1 hour.
(3) washing lotion configured washes plate 5 times, and button is dry.
(4) every enzyme-added 100ul in hole, continues at 37 DEG C of incubators and hatches 1 hour.
(5) washing lotion configured washes plate 5 times, adds substrate (each 50ul of A, B liquid).
(6) 37 DEG C of incubators hatch 30 minutes.
(7) every hole adds stop buffer 50ul, and microplate develops the color, and detects OD value by microplate reader.
(8) the OD value measured by protoplasm control quality testing and the OD value that measures of colour developing the third liver Quality Control thing are carried out t and are checked and compare, P=0.196 > 0.05, two groups of data are without obvious significant difference, illustrate that the Quality Control thing after this colouring method process can well maintain the Biochemistry laboratory course of Quality Control thing, the enzyme linked immunological that is applied to that can be stable is tested.For confirming that the Quality Control thing after improving does not produce any impact to Blood Transfusion Services's routine work, detect in Blood Transfusion Services the microplate of hepatitis C virus add improvement in the blank well reserved after Quality Control thing 10ul, enter in full-automatic FAME aftertreatment instrument with microplate and test one month, doing microplate number is altogether 178 plates, by improve after Quality Control thing institute's value and protoplasm control thing numeric ratio comparatively, (P=0.177>0.05, difference does not have statistical significance).Quality Control thing after improving is joined any hole in microplate at random together with detection sample, do microplate 20 pieces altogether, the value improving rear Quality Control thing gained is compared with protoplasm control thing OD value, (P=0.135>0.05, difference does not have statistical significance).
It should be noted last that, above embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to technical scheme of the present invention or equivalent replacement, and not departing from the spirit and scope of technical solution of the present invention, it all should be encompassed in the middle of right of the present invention.
Claims (4)
1. a Color-development quality control material, is characterized in that, described Color-development quality control material is that in common Quality Control thing, add concentration be that the coloring agent of 0.001g/ml ~ 1g/ml obtains; Described common Quality Control thing is AIDS virus Quality Control thing, the third liver Quality Control thing, syphilis Quality Control thing or hepatitis B Quality Control thing; When described common Quality Control thing is AIDS virus Quality Control thing, syphilis Quality Control thing or hepatitis B Quality Control thing, the concentration adding coloring agent is 0.1g/ml; When described common Quality Control thing is the third liver Quality Control thing, the concentration adding coloring agent is 0.2g/ml; Described coloring agent is famille rose, lemon yellow or sunset yellow.
2. Color-development quality control material according to claim 1, is characterized in that, in described Color-development quality control material, original Biochemistry laboratory course of common Quality Control thing can not change.
3. the application of the Color-development quality control material according to any one of claim 1 or 2, it is characterized in that, described Color-development quality control material may be used for enzyme-linked immunosorbent assay.
4. the application of Color-development quality control material according to claim 3, is characterized in that, described Color-development quality control material can ensure the stability of enzyme-linked immunosorbent assay.
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| CN105277686A (en) * | 2014-06-26 | 2016-01-27 | 赖平安 | Two diluent indicators in ELISA detection reagent kit |
| CN111638106B (en) * | 2020-06-09 | 2024-03-12 | 吉林基蛋生物科技有限公司 | Urine dry chemical analysis quality control object |
| CN113092775A (en) * | 2021-03-09 | 2021-07-09 | 中山生物工程有限公司 | Hepatitis B virus surface antibody detection kit and preparation method thereof |
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| CN1117586A (en) * | 1994-08-24 | 1996-02-28 | 张永新 | Hepatitis, AIDS, syphilis membranous type trigeminal immunoassay device and method |
| CN101097216A (en) * | 2007-06-18 | 2008-01-02 | 曹健荣 | Human immunodeficiency virus antibody confirmations reagent |
| CN102053154A (en) * | 2009-11-09 | 2011-05-11 | 深圳市宏信生物技术有限公司 | Kit for detecting autoimmune antibodies at throughput |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100537504B1 (en) * | 2003-01-20 | 2005-12-19 | 삼성전자주식회사 | An automated fluidic system for protein detection in biological samples |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1117586A (en) * | 1994-08-24 | 1996-02-28 | 张永新 | Hepatitis, AIDS, syphilis membranous type trigeminal immunoassay device and method |
| CN101097216A (en) * | 2007-06-18 | 2008-01-02 | 曹健荣 | Human immunodeficiency virus antibody confirmations reagent |
| CN102053154A (en) * | 2009-11-09 | 2011-05-11 | 深圳市宏信生物技术有限公司 | Kit for detecting autoimmune antibodies at throughput |
Non-Patent Citations (1)
| Title |
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| 自制人类免疫缺陷病毒抗体等质控血清及室内质控方法建立研究;柏居林等;《检验医学与临床》;20100228;第7卷(第3期);230-232 * |
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