CN212060271U - Antibody detection kit for detecting human brucella - Google Patents
Antibody detection kit for detecting human brucella Download PDFInfo
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- CN212060271U CN212060271U CN202020678040.2U CN202020678040U CN212060271U CN 212060271 U CN212060271 U CN 212060271U CN 202020678040 U CN202020678040 U CN 202020678040U CN 212060271 U CN212060271 U CN 212060271U
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Abstract
The utility model provides a detect antibody detect reagent box of people's brucellosis, comprises a box bod, the top of box body articulates there is the lid, the inside baffle that is provided with of box body, the baffle will the inner space of box body divide into two parts, deposits the district and test solution deposits the district for the detection card respectively, it has people's brucellosis to detect the card to deposit to be provided with in the district to detect the card, test solution deposits and is provided with sample diluent bottle and sample in the district and adds the burette. The detection kit of the utility model has simple operation, wide use, good specificity and high sensitivity, and is suitable for the auxiliary diagnosis of early acute infection and the on-site diagnosis of epidemiology investigation.
Description
Technical Field
The utility model belongs to the kit field especially relates to an antibody detection kit who detects human brucella.
Background
Brucellosis, also known as Malta fever or wavy fever, is a natural epidemic infectious disease of zoonosis caused by Brucella (also known as Brucella). The epidemic is wide, almost all over the world, and over 160 countries and regions have the epidemic of brucellosis. Brucellosis can infect humans, livestock and wild animals, resulting in significant economic losses and serious public health problems. The harm of human infected with brucellosis is more serious than that of domestic animal.
After people are infected with brucellosis, the brucellosis has no characteristic symptoms, is often converted into chronic diseases due to misdiagnosis and mistreatment, causes repeated attacks, has low cure rate, is difficult to cure for the whole life of patients, can lead delayed patients to lose labor capacity, can cause death seriously, and brings inestimable loss to individuals, families and society. Moreover, the disease has high incidence rate, complex disease symptoms, various organs can be infected, and the diagnosis is difficult.
Currently, the detection of brucella is mainly carried out in a laboratory, and the main methods comprise: (1) and (3) a separate culture method which is easily influenced by a culture method to cause false negative results. Moreover, the culture period is long, the operation is difficult, and the method is gradually eliminated. (2) The molecular biology method needs to synthesize specific primers and cooperate with corresponding instruments and equipment to perform detection and result analysis, and is not suitable for field application and rapid diagnosis. (3) The serological diagnosis method has the advantages of low specificity and sensitivity, complex operation, large serum consumption and low detection speed.
Therefore, the establishment of a brucellosis diagnosis kit which is simpler and more convenient to operate, low in cost, good in specificity, high in sensitivity, suitable for popularization, capable of carrying out single-part detection and more suitable for early acute infection auxiliary diagnosis and epidemiological investigation field application is urgently needed.
SUMMERY OF THE UTILITY MODEL
The to-be-solved problem of the utility model is to provide an antibody detection kit for detecting human brucella, this detection kit is easy and simple to handle, use extensively, the specificity is good, sensitivity is high, is suitable for early acute infection auxiliary diagnosis and epidemiology investigation on-the-spot diagnosis.
In order to solve the technical problem, the utility model discloses a technical scheme is: the utility model provides a detect antibody detect reagent box of people's brucella, includes the box body, the top of box body articulates there is the lid, the inside baffle that is provided with of box body, the baffle will the inner space of box body divide into two parts, deposits the district and test solution storage area for the detection card respectively, it has people's brucella detection card to set up in the detection card storage area, test solution is deposited and is provided with sample diluent bottle and sample and adds the burette in the district.
Furthermore, a second foam partition plate is arranged at the bottom of the test solution storage area, a first round hole and a second round hole are formed in the second foam partition plate, a sample diluent bottle is fixedly clamped in the first round hole, and a sample adding dropper is fixedly clamped in the second round hole.
Furthermore, a brucella detection card is arranged in the detection card storage area, 4 fixed clamping blocks are fixedly arranged on the inner wall of the detection card storage area and are respectively fixed at four corners of the detection card storage area, a fixed clamping cavity is formed between the 4 fixed clamping blocks, and the brucella detection card is arranged in the fixed clamping cavity.
Further, the first round hole and the second round hole are both provided with one.
Furthermore, 3 human brucella detection cards are arranged in the detection card storage area, 12 fixing clamping blocks are fixedly arranged on the inner wall of the detection card storage area, every four fixing clamping blocks form a fixing clamping cavity, and the 3 human brucella detection cards are respectively clamped and fixed in different fixing clamping cavities.
Further, the first round hole is provided with one, and the second round hole is provided with three.
Furthermore, a first foam partition plate and a third foam partition plate are arranged in the detection card storage area, the first foam partition plate is located at the bottom of the fixed clamping block, and the third foam partition plate is arranged above the human Brucella detection card.
Further, the human brucella detection card comprises a card shell and a detection test strip arranged in the card shell; the detection test strip comprises a reaction plate, a sample pad, a gold conjugate pad, a reaction membrane and coarse fiber filter paper, wherein the sample pad, the gold conjugate pad, the reaction membrane and the coarse fiber filter paper are arranged on the reaction plate in a clinging manner, one end of the sample pad covers the upper side of one end of the gold conjugate pad, the other end of the gold conjugate pad covers the upper side of one end of the reaction membrane, and one end of the coarse fiber filter paper covers the upper side of the other end of the reaction membrane; the reaction membrane is provided with a first detection line, a second detection line and a quality control line, and the first detection line, the second detection line and the quality control line are sequentially arranged from a near gold conjugate pad to a near coarse fiber filter paper.
Further, the card shell comprises a first card shell and a second card shell, a sample adding hole is formed in the first card shell, an observing hole is formed in the second card shell, the sample adding hole is located on the upper side of the sample pad, and the observing hole is located above the first detecting line, the second detecting line and the quality control line.
Further, in the human brucella detection card, the gold conjugate pad is coated with a recombinant brucella antigen labeled with colloidal gold, the first detection line is coated with a mouse anti-human lgM monoclonal antibody, the second detection line is coated with a mouse anti-human lgG monoclonal antibody, and the quality control line is coated with a mouse anti-brucella antibody.
Furthermore, locking devices are arranged on the box cover and the box body in a matched mode.
The utility model has the advantages and positive effects that:
the human brucella detection card in the kit has the characteristics of rapidness, simplicity, convenience, no need of special equipment, high accuracy, high sensitivity and the like, and the whole operation time is only 15-20 minutes. The clinical detection has important application value for early diagnosis of patients; and is suitable for the auxiliary diagnosis of early acute infection and the epidemiological investigation.
Drawings
FIG. 1 is a front view of an antibody detection kit for detecting human Brucella according to the present invention;
FIG. 2 is a sectional view of embodiment 1 of the antibody detection kit for detecting human Brucella according to the present invention;
FIG. 3 is a cross-sectional view A-A of FIG. 2;
FIG. 4 is a schematic structural diagram of embodiment 1 of the antibody detection kit for detecting human Brucella;
FIG. 5 is a sectional view of embodiment 2 of the antibody detection kit for detecting human Brucella according to the present invention;
FIG. 6 is a cross-sectional view A-A of FIG. 5;
FIG. 7 is a schematic structural diagram of embodiment 2 of the antibody detection kit for detecting human Brucella according to the present invention;
FIG. 8 is a schematic structural diagram of a card shell in an antibody detection kit for detecting human Brucella according to the present invention;
FIG. 9 is a schematic structural diagram of a test strip in an antibody detection kit for detecting human Brucella according to the present invention;
FIG. 10 is a schematic structural diagram of a fixed block in an antibody detection kit for detecting human Brucella according to the present invention;
in the figure: 1-box body, 2-box cover, 3-clapboard, 4-detection card storage area, 5-test solution storage area, 6-brucella abortus detection card, 61-card shell, 611-first card shell, 6111-sample adding hole, 612-second card shell, 6121-observation hole, 62-detection test paper strip, 621-reaction plate, 622-sample pad, 623-gold conjugate pad, 624-reaction membrane, 6241-first detection line, 6242-second detection line, 6243-quality control line, 625-coarse fiber filter paper, 7-sample diluent bottle, 8-sample adding dropper, 9-fixed fixture block, 10-fixed card cavity, 11-first foam clapboard, 12-second foam clapboard, 121-first round hole, 122-second round hole, 13-third foam spacer.
Detailed Description
The following detailed description of the embodiments of the present invention will be made with reference to the accompanying drawings.
Example 1:
as shown in fig. 1 to 4 and fig. 8 to 10: an antibody detection kit for detecting human brucella comprises a box body 1, wherein a box cover 2 is hinged to the top of the box body 1, a partition plate 3 is arranged inside the box body 1, the partition plate 3 divides the inner space of the box body 1 into two parts, namely a detection card storage area 4 and a test solution storage area 5, and the partition plate 3 is used for separating the detection card storage area 4 from the test solution storage area 5 so as to be convenient to take; the detection card storage area 4 is internally provided with a brucella detection card 6, and the test solution storage area 5 is internally provided with a sample diluent bottle 7 and a sample adding dropper 8. The sample diluent 7 is filled with a sample diluent, namely a phosphate buffer, and the sample adding dropper 8 is used for adding a sample during detection.
In this embodiment, a second foam partition 12 is disposed at the bottom of the test solution storage area 5, a first circular hole 121 and a second circular hole 122 are formed in the second foam partition 12, a sample diluent bottle 7 is fixedly clamped in the first circular hole 121, and a sample adding dropper 8 is fixedly clamped in the second circular hole 122. Wherein, the first round hole 121 and the second round hole 122 are both provided with one. A second foam spacer 12 is provided for holding the specimen dilution bottle 7 and the specimen addition dropper 8.
In this embodiment, a brucella detection card 6 is disposed in the detection card storage area 4, 4 fixing blocks 9 are fixedly disposed on the inner wall of the detection card storage area 4, and are respectively fixed at four corners of the detection card storage area 4, a fixing card cavity 10 is formed between the 4 fixing blocks 9, and the brucella detection card 6 is disposed in the fixing card cavity 10. The fixing block 9 is arranged to form a fixing block cavity 10, so that the detection card can be fixed conveniently and prevented from shifting.
In this embodiment, a first foam partition plate 11 and a third foam partition plate 13 are disposed in the detection card storage area 4, the first foam partition plate 11 is located at the bottom of the fixed fixture block 9, and the third foam partition plate 13 is disposed above the human brucella detection card 6. Set up first foam baffle 11 and play the effect that separates wet heat-proof, set up third foam baffle 13 and can prevent that the detection card from removing, play the cushioning effect simultaneously, in the kit transportation, play the cushioning effect when its atress again.
Further, the human brucella detection card 6 comprises a card shell 61 and a detection test strip 62 arranged in the card shell 61; the card shell 61 is used for packaging and fixing the test strip 62; the detection test strip comprises a reaction plate 621, a sample pad 622, a gold conjugate pad 623, a reaction membrane 624 and a coarse fiber filter paper 625, wherein the reaction membrane 624 is a nitrocellulose membrane; wherein, the sample pad 622, the gold conjugate pad 623, the reaction membrane 624 and the coarse fiber filter paper 625 are closely arranged on the reaction plate 621, one end of the sample pad 622 covers the upper side of one end of the gold conjugate pad 623, the other end of the gold conjugate pad 623 covers the upper side of one end of the reaction membrane 624, and one end of the coarse fiber filter paper 625 covers the upper side of the other end of the reaction membrane 624; the reaction membrane 624 is provided with a first detection line 6241, a second detection line 6242 and a quality control line 6243, and the first detection line 6241, the second detection line 6242 and the quality control line 6243 are sequentially arranged from the adjacent gold conjugate pad 623 to the adjacent coarse fiber filter paper 625. At the time of detection, a sample is added from the sample pad 622, and then the sample is diffused from the sample pad 622 to the gold conjugate pad 623 and then into the reaction membrane 624 in sequence for diffusion.
Further, the card shell 61 is composed of a first card shell 611 and a second card shell 612, a sample adding hole 6111 is formed in the first card shell 611, an observation hole 6121 is formed in the second card shell 612, the sample adding hole 6111 is located on the upper side of the sample pad 622, and the observation hole 6121 is located above the first detection line 6241, the second detection line 6242 and the quality control line 6243. The sample adding hole 6111 is used for adding a sample, and the observation hole 6121 is used for observing whether the first detection line 6241, the second detection line 6242 and the quality control line 6243 change color or not.
Further, in the human brucella detection card 6, the gold conjugate pad 623 is coated with a recombinant brucella antigen labeled with colloidal gold, the first detection line 6241 is coated with a mouse anti-human lgM monoclonal antibody, the second detection line 6242 is coated with a mouse anti-human lgG monoclonal antibody, and the quality control line 6243 is coated with a mouse anti-brucella antibody.
When an IgM antibody positive sample is detected, the human brucella IgM antibody in the sample can be combined with the recombinant brucella antigen marked by the colloidal gold to form an immune complex, and the complex and the sample flow forwards in the reaction membrane 164 due to the chromatography; when the compound passes through the first detection line 1641, the compound is combined with the coated mouse anti-human IgM monoclonal antibody to form 'colloidal gold-recombinant brucella antigen-human brucella IgM-mouse anti-human IgM monoclonal antibody' for agglutination and color development. When an IgG antibody positive sample is detected, the human Brucella IgG antibody in the sample can be combined with the recombinant Brucella antigen marked by the colloidal gold to form an immune complex, and the complex and the sample flow forwards in the reaction membrane 164 due to chromatography; when the complex passes through the second detection line 1642, the complex is combined with the coated mouse anti-human IgG monoclonal antibody to form the colloidal gold-recombinant brucella antigen-human brucella IgG-mouse anti-human IgG monoclonal antibody, and agglutination and color development are realized. The residual colloidal gold labeled recombinant brucella antigen is combined with the mouse anti-brucella antibody coated by the quality control line 1643 to agglutinate and develop color, and when a negative sample is detected, the sample does not contain the combination of human brucella IgM/IgG antibody, so that an immune complex cannot be formed, and the color can be developed only at the position of the quality control line 1643.
Furthermore, locking devices are arranged on the box cover 2 and the box body 1 in a matched mode. The locking device is of any structure capable of fixing and separating the box cover and the box body, for example, the locking device can be an upper lock catch and a lower lock catch which are matched with each other, the upper lock catch is arranged on the box cover 2, the lower lock catch is arranged on the box body 1, and when the box cover 2 is locked with the box body 1, the upper lock catch and the lower lock catch are locked.
Example 2:
as shown in fig. 5-10, compared with embodiment 1, in this embodiment, 3 human brucella detection cards 6 are disposed in the detection card storage area 4, 12 fixing clips 9 are fixedly disposed on the inner wall of the detection card storage area 4, wherein every four fixing clips 9 form a fixing clip cavity 10, and 3 human brucella detection cards 6 are respectively fixed in different fixing clip cavities 10.
In this embodiment, there are one first circular hole 121 and three second circular holes.
Compared with the embodiment 1, the difference is that the kit of the embodiment is equipped with three human brucella detection cards 6, three samples can be tested, and the kit is more suitable for being carried outside compared with a single sample, so as to reduce the occupied space.
In the utility model, the preparation process of the specific human brucella detection card 6 is as follows:
1. preparation of reaction film 624: diluting the mouse anti-human lgM monoclonal antibody and the mouse anti-human lgG monoclonal antibody to 1mg/mL by using coating diluent; diluting the mouse anti-brucella antibody to 2mg/mL by using a coating diluent; spraying the two coating solutions onto the reaction film 4 by a film dispenser with a spraying amount of 0.1 μ L/mm to form a first detection line 6241, a second detection line 6242 and a quality control line 6243 respectively; the coated reaction membrane 624 was dried at 37 ℃ for 3 hours and stored at room temperature. Wherein the coating diluent is phosphate buffer.
2. Preparation of gold conjugate pad 623: and coupling the recombinant brucella antigen with colloidal gold to prepare a colloidal gold-labeled recombinant brucella antigen solution. The colloidal gold-labeled recombinant brucella antigen solution was adjusted to a concentration of 10 μ g/mL, 1.5 mL/strip, the gold conjugate pad 623 was soaked, gold-plated, dried at 37 ℃ for 3 hours, and stored at room temperature. The preparation method of the colloidal gold labeled recombinant Brucella antigen comprises the following steps: regulating the pH value of the colloidal gold to 8.5-9.0 by using 0.1M potassium carbonate solution; adding 10 microgram/mL recombinant Brucella antigen, and stirring for 40 minutes; thirdly, adding 10 percent of bovine serum albumin to the final concentration of 1 percent, and stirring for 61 minutes; fourthly, centrifuging at 12000rpm/min for 61 minutes at 4 ℃, and carefully removing supernatant; adding 1/2 volume colloidal gold conjugate diluent of original volume (original volume is calculated by colloidal gold volume); the colloidal gold conjugate diluent is prepared by dissolving 5.372g of disodium hydrogen phosphate, 0.78g of sodium dihydrogen phosphate, 5g of bovine serum albumin, 8.5g of sodium chloride and 0.5g of casein in 1L of deionized water and uniformly mixing.
3. Cutting and assembling: the reaction membrane 624, the gold conjugate pad 623, the coarse fiber filter paper 625 and the sample pad 622 are sequentially attached to the reaction plate 621, and then cut into a test strip 62 having a width of 4mm by a cutter, and then the test strip 62 is assembled in the card case 61, thereby obtaining the final human brucella detection card 6.
The procedure for detection using the human brucella detection card 6 was as follows:
1. taking 10 μ L of serum, plasma or 20 μ L of whole blood sample, directly adding into the sample adding hole 6111, and adding 100 μ L of sample diluent (about 2-3 drops); the whole blood is collected by fingertips or veins, and the whole blood sample is not preserved after being collected, and is used immediately after being collected; collecting serum sample by vein according to conventional method; the plasma sample can be treated by heparin, sodium citrate and EDTA; the sample measured in 5 days of the serum or plasma sample can be stored at 4 ℃;
2. observing the color changes of the first detection line 6241, the second detection line 6242 and the quality control line 6243 at the observation hole 6121, judging and reading the result within 61-20 minutes, and invalidating the detection result after 20 minutes;
3. and (3) judging a detection result: when a red strip appears at the position of the quality control line 6243, the result is negative; IgM is positive when two red bands appear at the positions of the quality control line 6243 and the first detection line 6241; IgG is positive when two red strips appear at the positions of the quality control line 6243 and the second detection line 6242; detection is invalid when no red band appears at the position of the control line 6243.
As can be seen from the above, the human Brucella detection card 6 in the kit has the characteristics of rapidness, simplicity, convenience, no need of special equipment, high accuracy and sensitivity and the like, and the whole operation time is only 15-20 minutes. The kit has important application value for early diagnosis of patients and clinical detection. And is suitable for the auxiliary diagnosis of early acute infection and the epidemiological investigation.
The above description is provided for the preferred embodiments of the present invention, but the above description is only for the preferred embodiments of the present invention, and should not be construed as limiting the scope of the present invention. All the equivalent changes and improvements made according to the application scope of the present invention should still fall within the patent coverage of the present invention.
Claims (10)
1. The utility model provides an antibody detection kit for detecting human brucella, includes box body (1), the top of box body (1) articulates there is lid (2), its characterized in that: the utility model discloses a reagent bottle box, including box body (1), baffle (3) will the inner space of box body (1) divide into two parts, deposit district (4) and test solution and deposit district (5) for the detection card respectively, it has brucella detection card (6) to be provided with in detection card deposit district (4), it adds burette (8) to be provided with sample dilution liquid bottle (7) and sample in test solution deposit district (5).
2. The antibody detection kit for detecting human brucella as claimed in claim 1, wherein: the bottom of the test solution storage area (5) is provided with a second foam partition plate (12), the second foam partition plate (12) is provided with a first round hole (121) and a second round hole (122), a sample diluent bottle (7) is fixedly clamped in the first round hole (121), and a sample adding dropper (8) is fixedly clamped in the second round hole (122).
3. The antibody detection kit for detecting human brucella as claimed in claim 2, wherein: the detection card storage area (4) is internally provided with a brucella detection card (6) for one person, the inner wall of the detection card storage area (4) is fixedly provided with 4 fixed clamping blocks (9) which are respectively fixed at four corners of the detection card storage area (4) and 4, a fixed card cavity (10) is formed between the fixed clamping blocks (9), and the brucella detection card (6) for one person is arranged in the fixed card cavity (10).
4. The antibody detection kit for detecting human brucella as claimed in claim 3, wherein: the first round hole (121) and the second round hole (122) are both provided with one.
5. The antibody detection kit for detecting human brucella as claimed in claim 2, wherein: the detection card storage area (4) is internally provided with 3 Brucella detection cards (6), the inner wall of the detection card storage area (4) is fixedly provided with 12 fixed clamping blocks (9), every four fixed clamping blocks (9) form a fixed card cavity (10), and 3 Brucella detection cards (6) are respectively clamped and fixed in different fixed card cavities (10).
6. The antibody detection kit for detecting human brucella as claimed in claim 5, wherein: the number of the first round holes (121) is one, and the number of the second round holes is three.
7. The antibody detection kit for detecting human brucella as claimed in claim 3 or 5, wherein: the detection card storage area (4) is internally provided with a first foam partition plate (11) and a third foam partition plate (13), the first foam partition plate (11) is located at the bottom of the fixed clamping block (9), and the third foam partition plate (13) is arranged above the human Brucella detection card (6).
8. The antibody detection kit for detecting human brucella as claimed in claim 1, wherein: the human Brucella detection card (6) comprises a card shell (61) and a detection test strip (62) arranged in the card shell (61); the detection test strip comprises a reaction plate (621), a sample pad (622), a gold conjugate pad (623), a reaction membrane (624) and coarse fiber filter paper (625), wherein the sample pad (622), the gold conjugate pad (623), the reaction membrane (624) and the coarse fiber filter paper (625) are closely arranged on the reaction plate (621), one end of the sample pad (622) covers the upper side of one end of the gold conjugate pad (623), the other end of the gold conjugate pad (623) covers the upper side of one end of the reaction membrane (624), and one end of the coarse fiber filter paper (625) covers the upper side of the other end of the reaction membrane (624); the reaction membrane (624) is provided with a first detection line (6241), a second detection line (6242) and a quality control line (6243), and the first detection line (6241), the second detection line (6242) and the quality control line (6243) are sequentially arranged from an adjacent gold conjugate pad (623) to an adjacent coarse fiber filter paper (625).
9. The antibody detection kit for detecting human brucella as claimed in claim 8, wherein: the clamping shell (61) is formed by clamping a first clamping shell (611) and a second clamping shell (612), a sample adding hole (6111) is formed in the first clamping shell (611), an observation hole (6121) is formed in the second clamping shell (612), the sample adding hole (6111) is located on the upper side of the sample pad (622), and the observation hole (6121) is located above the first detection line (6241), the second detection line (6242) and the quality control line (6243).
10. The antibody detection kit for detecting human brucella as claimed in claim 8, wherein: in the human brucella detection card (6), a gold conjugate pad (623) is coated with a recombinant brucella antigen labeled with colloidal gold, a first detection line (6241) is coated with a mouse anti-human lgM monoclonal antibody, a second detection line (6242) is coated with a mouse anti-human lgG monoclonal antibody, and a quality control line (6243) is coated with a mouse anti-brucella antibody.
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| CN202020678040.2U CN212060271U (en) | 2020-04-28 | 2020-04-28 | Antibody detection kit for detecting human brucella |
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| CN202020678040.2U CN212060271U (en) | 2020-04-28 | 2020-04-28 | Antibody detection kit for detecting human brucella |
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