CN209264734U - Brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream - Google Patents

Abstract

The utility model relates to brucella antibody test colloidal gold colloidal gold detection test paper strips in cattle and sheep cream, it is characterised in that: including micropore cup and the sealing element being set on micropore cup and test strips；Colloidal gold-SPA the compound of freeze-drying is placed in the micropore cup；The test strips are equipped with LPS detection line T and rabbit-anti SPA polyclonal antibody nature controlling line C.Brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream provided by the utility model, it is short with detection time, stability is good, it is easy to operate, be not necessarily to other instruments, do not require testing staff that there is specialized capability yet, testing result is intuitive and reliable simultaneously, and the detection of high-volume sample is carried out suitable for base's milking station and quality testing department.

Description

Brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream

Technical field

The utility model relates to colloidal gold colloidal gold detection test paper strip fields, examine more particularly, to brucella antibody in cattle and sheep cream Survey colloidal gold colloidal gold detection test paper strip.

Background technique

Brucella is a kind of Gram-negative bacteria of cytozoicus.It can be divided according to its pathogenic and host's preferendum It is 7 kinds, 20 biological.That wherein infects people has ox, sheep, pig, dog and marine species.Sheep kind it is brucellar it is pathogenic most By force, clinically maximum with the meaning of the brucellosis of ox, sheep, pig.Brucellosis is the people, animal as caused by brucella Illness altogether.

Brucellar harm is larger, people suffer from brucella it is clinical after being ill be mainly shown as that fever, hidrosis, whole body are weary Power, arthralgia, and can lead to deformity, women easily miscarries, and male is then susceptible to suffer from orchitis and leads to the forfeiture of fecundity.Because of it It is similar with cold symptoms and ignored by people, if cannot timely symptomatic treatment, severe can be formed or be transferred to chronic phase, can different journeys Degree ground influences the physical and mental health and labour capacity of people.Clinical symptoms are mainly shown as after being ill for bovine livestock, sheep, pig infection brucella Reproductive organs, fetal membrane, Various Tissues organ irritation, necrosis form granuloma, and male animal is susceptible to suffer from orchitis, epididymitis leads to male animal Infertility, dam easily leads to miscarriage and infertile, weak tire or stillborn foetus etc..Because the disease is the total illness of people and animals' easy infection and to the mankind Health, livestock-raising and food safety bring huge harm, therefore the disease is our country is listed as two class infectious diseases.

The traditional immunization method of detection brucellosis includes the red plate agglutination experiment (RBPT) of tiger, plate agglutination at present Test (PAT), standard test tube agglutination experiment several traditional serological diagnostic methods such as (SAT) and complement fixation test (CFT). The milk of external cattle farm production is cooked oligomer experiment.The above-mentioned detection method operation about brucella antibody is more numerous It is trivial, it takes long time, and not intuitive enough as the result is shown, meanwhile, above-mentioned detection method also requires test experience personnel that must have phase The experimental skills answered can just access accurate detection result.In this case, base's milking station and quality testing department are not particularly suited for The screening and detection of high-volume sample.

Utility model content

In view of this, the utility model is intended to provide one kind can be used in quickly detecting in cattle and sheep cream whether contain cloth Lu Shi The colloidal gold colloidal gold detection test paper strip of bacillus, to solve, existing traditional immunization detection method is cumbersome, takes long time, as the result is shown The not intuitive and demanding problem of implementation.

The technical solution adopted in the utility model is as follows:

Cattle and sheep cream in brucella antibody test colloidal gold colloidal gold detection test paper strip, including test strips and with test strips cooperate The micropore cup used is provided with sealing element on the micropore cup；Colloidal gold-the SPA that freeze-drying is placed in the micropore cup is compound Object；The test strips are equipped with LPS (Lipopolysaccharide, lipopolysaccharides) detection line T and rabbit-anti SPA (Staphylocoal Protein A, staphylococcal protein A) polyclonal antibody nature controlling line C.

Further, the LPS detection line T and rabbit-anti SPA polyclonal antibody nature controlling line C are arranged parallel to each other.

Further, the quantity of the rabbit-anti SPA polyclonal antibody nature controlling line C is 1.

Further, the test strips include bottom plate, and the test strips include bottom plate, and successively linking fits in bottom plate On sample pad, nitrocellulose filter and blotting paper.

Further, the sample pad is glass fiber sample pad.

Further, the bottom plate is PVC board.

Further, the micropore cup includes cup plate；Several indents are provided on the cup plate and for placing colloid The cup-like trough of gold-SPA compound；The sealing element is to be set to glass plate surface and for sealing the opening position of cup-like trough Any one of sealed membrane or micropore lid.

Further, when the sealing element be micropore lid when, the micropore lid include it is equal with cup-shaped slot number and with The one-to-one seal closure of cup-like trough；Each seal closure lower end is equipped with the protrusion isometrical with the opening of cup-like trough, so that envelope Lid can be sticked in the opening of cup-like trough and seal cup-like trough.

In conclusion compared with prior art, by adopting the above-described technical solution, the beneficial effects of the utility model It is:

(1) high sensitivity of colloidal gold colloidal gold detection test paper strip provided by the utility model: it is with colloid gold label SPA, colloid Gold label effect is preferable, and the colloidal gold-SPA marked is added in micropore cup after being diluted with freeze-drying liquid and is lyophilized.It can be with when detection Sample and Sample dilution is first added in micropore cup, antibody and colloidal gold-SPA in sample can so sufficiently reacted, Then test strips are inserted into, to substantially increase the sensitivity of detection reagent；It tests international Bang's bacillus positive criteria The sensitivity of serum can reach 0.1lU；

(2) specificity of colloidal gold colloidal gold detection test paper strip provided by the utility model is good: through testing, this test strips detects Yang Yuan Salmonella positive serum, ox source salmonella positive serum, sheep source colon bacillus 0157 positive serum, ox source Escherichia coli 0157 positive serum, Mycobacterium bovis by force positive serum, Rough Anti-Brucella positive serum when be feminine gender；

(3) colloidal gold colloidal gold detection test paper strip provided by the utility model is easy to operate, and as a result interpretation is quick: only needing absorption 10 The fresh milk sample of μ L is added in micropore cup, and the Sample dilution that 100 μ L are then added makes sample in micropore cup and freeze-drying After colloidal gold-SPA is sufficiently mixed, result can be directly observed after being inserted into 5~8min of test strips；

(4) the display testing result of colloidal gold colloidal gold detection test paper strip provided by the utility model is vivid, intuitive, accurate；With inspection Whether test paper slip T line develops the color as negative and positive label, while judging that test strip is with the presence or absence of Quality Control elder generation C line No failure provides effective guarantee to testing result；When test strip T line does not develop the color the colour developing of C line, indicate to be free of in sample There is Brucella antibody；When test strip T line and C line develop the color, indicate to contain Brucella antibody in sample；Once inspection Test paper slip C line does not develop the color, and indicates test strip failure；

(5) cost of colloidal gold colloidal gold detection test paper strip provided by the utility model is low, does not need in use process any mating Analysis Instrument equipment and reagent, without without acquiring animal blood and separation separation serum, layman can be carried out Detection operates and obtains accurate result；Thus, it is easy to which base promotes, the big base's detection scene of detection demand can satisfy, Such as farm, health and epidemic prevention station, base's stock units, practicability with higher.

Detailed description of the invention

Fig. 1 is the structural plan of test strips described in the utility model

Fig. 2 is a kind of structural schematic diagram of micropore cup described in the utility model

Fig. 3 is a kind of structural schematic diagram of micropore lid described in the utility model

Fig. 4 is that colloidal gold colloidal gold detection test paper strip provided by the utility model determines that sample to be tested testing result contains brucella The schematic diagram of antibody

Fig. 5 is that colloidal gold colloidal gold detection test paper strip provided by the utility model determines sample to be tested testing result without brucella The schematic diagram of antibody

Fig. 6 is colloidal gold colloidal gold detection test paper strip failure provided by the utility model or expired schematic diagram

In the figure, it is marked as 1- blotting paper, 2- rabbit-anti SPA polyclonal antibody nature controlling line C, 3-LPS detection line T, 4- nitric acid are fine Tie up plain film, 5- sample pad, 6- glasss of plates, 7- cup-like trough, 8- micropore lid, 9- bottom plate.

In Fig. 4, a and b indicate that colloidal gold colloidal gold detection test paper strip determines that sample to be tested testing result contains Brucella antibody Two kinds of situations, it is obvious not as good as in b that the difference of the two is that the LPS detection line T in a develops the color, and typicallys represent to be measured representated by a Brucella antibody content is less than sample to be tested representated by b in sample.

In Fig. 6, the difference of c and d are that the LPS detection line T in c develops the color, and LPS detection line T does not develop the color in d, but c and d Middle rabbit-anti SPA polyclonal antibody nature controlling line C do not develop the color, to illustrate to indicate test strip failure or expired be likely to occur Two kinds of testing results.

Specific embodiment

All features disclosed in this specification can be with any other than mutually exclusive feature and/or step Mode combines.

It elaborates below with reference to Fig. 1~6 pair the utility model.

Embodiment 1

For the ease of the quick and precisely detection of brucella antibody in development at the basic level ox, sheep cream, referring to Figures 1 and 2, this Embodiment provides brucella antibody test colloidal gold colloidal gold detection test paper strip in a kind of cattle and sheep cream, including test strips and and test paper The micropore cup that item is used cooperatively is provided with sealing element on the micropore cup, the colloidal gold-of freeze-drying is placed in the micropore cup SPA compound；The test strips are equipped with LPS detection line T3 and rabbit-anti SPA polyclonal antibody nature controlling line C2.

Wherein, see Fig. 1, the test strips include bottom plate 9, and the test strips include bottom plate 9, and successively linking fits in Sample pad 5, nitrocellulose filter 4 and blotting paper 1 on bottom plate 9.See that Fig. 2, the micropore cup include cup plate 6；On the cup plate 6 It is provided with several indents and the cup-like trough 7 for placing colloidal gold-SPA compound；The sealing element is to be set to 6 table of glass plate Face and any one of the sealed membrane for being used to make the opening position of cup-like trough 7 to seal or micropore lid 8.Meanwhile the micropore cup Material can be but be not limited to the plastics such as medical grade PVC, PC.

For above-mentioned colloidal gold colloidal gold detection test paper strip, expansion explanation is carried out below.

1, colloidal gold-SPA compound the preparation method is as follows:

1) preparation of colloidal gold

Add 100mL ultrapure water into the 300mL triangular flask of wash clean, triangular flask is then placed in heating magnetic stirring apparatus On be heated to boiling, then phase triangular flask in be added 1mL 1% chlorogold solution, center boiling after be added bis- water lemon of 1.5mL Lemon acid trisodium continues 15min after being heated to boiling and stops heating, ultrapure water is added after restoring to room temperature and restores to original volume.4℃ It saves backup.

2) label of SPA colloidal gold

The pH of colloidal gold solution is adjusted to 6.0 with the potassium carbonate of 0.1mol/L under magnetic stirring by colloidal gold solution.By every The amount that 16 micrograms are added in milliliter colloidal gold is slowly added to SPA.It is stirred at room temperature and 10% BSA solution is added after 60min to end Concentration is 1%, continues to stir 60min at room temperature.After stirring stops, it being centrifuged under the conditions of 4 DEG C with the revolving speed of 4500r/min 20min collects centrifuge tube bottom precipitation, and upper layer colloidal gold solution continues be centrifuged 30min with the revolving speed of 8500r/min, again Collect centrifuge tube bottom precipitating, the discarded recycling of supernatant.The colloidal gold collected twice is precipitated with redissolution liquid according to colloid when label It redissolves the 1/20 of gold solution volume.4 DEG C save backup.

Wherein, redissolving liquid is the PB buffer that 20mM, pH are 7.4, and wherein BSA content is 1%, polysorbas20 0.3%, PVP40 is 1%, trehalose 3%, sucrose 5%.

2, the compound colloidal gold-SPA compound being prepared into micropore cup of the colloidal gold-SPA marked among the above is lyophilized The method of powder are as follows:

After the colloidal gold-SPA compound marked is diluted 14 times with freeze-drying liquid, dilution volume and amount are with each micropore cup Subject to 70 μ L of middle addition.Wherein, the PB buffer that freeze-drying liquid level 0.02mol/L, pH is 7.4, wherein BSA content is 1%, caramel 2%, trehalose 3%, polysorbas20 0.05%, dextrin 1%.It will be added to after above-mentioned dilution by the amount of each 70 μ L of micropore cup Then micropore cup equipped with above-mentioned colloidal gold-SPA compound is put into vacuum freeze drier, in -40 DEG C of item by micropore cup Start vacuum pump under part, after pre-freeze 5h to be vacuumized；Wherein, the temperature and setting time vacuumized in engineering can be -35 DEG C, 4h, -30 DEG C, 4h, -25 DEG C, 3h, -20 DEG C, 2h, -10 DEG C, 1h, -5 DEG C, 1h, 0 DEG C, 1h, 10 DEG C, 2h, 20 DEG C, 5h, 24 ℃,10h.After the completion of after freeze-drying, by the micropore cup being lyophilized after taking out in vacuum drier, humidity can be put into less than 30% Assembling room, then protected under the conditions of being placed in 4 DEG C using sealing element by after all cup-like trough 7 on micropore cup all sealing It deposits spare.

Wherein, the method for sealing element is set on micropore cup specifically:

When sealing element is sealed membrane, the sealed membrane is pressed together on the top of glass plate 6 from top to bottom under heat, infuses simultaneously Position between meaning cup-like trough 7 should also be as hot pressing pressing, so that the equal autonomous closure of the opening of each cup-like trough 7, what is used Remove the sealed membrane in the cup-like trough 7 for detection, such to be advantageous in that, cost of sealed membrane as desired in the process It is lower, and each cup-like trough 7 is allowed to be used alone.

When sealing element is micropore lid 8, it can be but be not limited to the plastics such as medical grade PVC, PC, and structure can be divided into Two kinds of situations.First, micropore lid 8 is independent design, i.e., being individually fitted into each cup-like trough 7 has to be used for the micro- of its sealing Port lid 8, advantage and the scheme of use sealed membrane are roughly the same, are made simultaneously because micropore lid 8 is usually selected compared with duroplasts, institute The sealing played is more reliable, can also more efficiently prevent from external force destruction, but cost can increase.Second, such as Fig. 3, institute There is micropore lid 8 to be integrated on one block of plate, enables to all cup-like troughs 7 to open simultaneously or close, the benefit being arranged in this way It is, assembling and using more convenient, operating efficiency is higher, while micropore lid 8 falls when greatly reducing micropore 8 independent design of lid The risk fallen.

3, the method for coating of sample pad is as follows:

The selection of sample pad can be, but not limited to the membranaceous sample pad of glass fibre material.

By taking glass fiber sample pad as an example, after impregnating 5min in its sample pad treatment fluid, in 37 DEG C of forced air drying It is sealed after drying in case spare.Wherein, sample pad treatment fluid is the PHS solution that 0.01mol/L, pH are 8.0, wherein BAS Content be 0.1%, the content of Triton X-100 content is 0.5%.

4, dotey's detection line and nature controlling line are as follows on nitrocellulose:

LPS to concentration 1mg/mL is diluted with the PBS solution that 0.01mol/L, pH are 7.4, is drawn by the amount of 1 μ L/cm in nitric acid On tunica fibrosa.1 μ L/ is pressed after diluting rabbit-anti SPA polyclonal antibody to 1.5mg/mL with the PBS solution that 0.01mol/L, pH are 7.4 On the amount language nitrocellulose filter 4 of cm.It is parallel to each other between two scribing line and is divided into 5mm.Film after coating is in 37 DEG C of drums Drying is sealed spare after drying overnight in wind drying machine.

5, the assembling of test strips

In composing room of the humidity less than 30%, by the sample pad 5 prepared, the nitrocellulose filter 4 being coated with, inhale Water paper 1 is successively pasted on PVC bottom plate 9, and the test strips for being then cut into 4mm wide with cutting machine are spare.

6, the assembling of finished product

Assembled micropore cup and the test strips cut are put into equipped with dry in composing room of the humidity less than 30% Assembling is sealed in the aluminium foil bag of drying prescription.Brucella antibody test colloidal gold detection in assembled finished product, as cattle and sheep cream Test strips, the test strips save under the conditions of 4 DEG C.

The application method of brucella antibody test colloidal gold colloidal gold detection test paper strip is as follows in cattle and sheep cream in the present embodiment:

In a cup-like trough 7 for taking the fresh ox of 10 μ L or sheep cream to be added on micropore cup with pipettor, 100 are then taken again The physiological saline of μ L is added in cup-like trough 7 to be aspirated 5 times repeatedly, so that fresh milk and the colloidal gold-SPA in cup-like trough 7 are compound Object freeze-dried powder mixes well.One end that taking-up test strips post sample pad 5 is inserted into cup-like trough 7, after waiting in 5~10min Interpretation result.

7, the interpretation of testing result

Referring to fig. 4, when red stripes occur in the C line of test strips, T line, show that detected fresh milk contains cloth Shandong Salmonella antibody；Referring to Fig. 5, there are red stripes at the only C line of test strips, shows detected fresh milk without containing cloth Shandong Salmonella antibody；Referring to Fig. 6, at the C line, T line of test strips equal redfree band occur or the C line of test strips at occur Red line band and at T line when redfree band, then show that test strips are no longer valid or expired.

It is as described above the embodiments of the present invention.The utility model is not limited to above embodiment, anyone It should learn the structure change made under the enlightenment of the utility model, it is all that there is same or similar skill with the utility model Art scheme, each falls within the protection scope of the utility model.

Claims (8)

1. cattle and sheep cream in brucella antibody test colloidal gold colloidal gold detection test paper strip, it is characterised in that: including test strips and with The micropore cup that test strips are used cooperatively is provided with sealing element on the micropore cup, the colloid of freeze-drying is placed in the micropore cup Gold-SPA compound；The test strips are equipped with LPS detection line T (3) and rabbit-anti SPA polyclonal antibody nature controlling line C (2).

2. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 1, feature exist In: the LPS detection line T (3) and rabbit-anti SPA polyclonal antibody nature controlling line C (2) are arranged parallel to each other.

3. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 2, feature exist In: the quantity of the rabbit-anti SPA polyclonal antibody nature controlling line C (2) is 1.

4. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 1, feature exist In: the test strips include bottom plate (9), and are successively connected the sample pad (5) fitted on bottom plate (9), nitrocellulose filter (4) and blotting paper (1).

5. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 4, feature exist In: the sample pad (5) is glass fiber sample pad (5).

6. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 4, feature exist In: the bottom plate (9) is PVC board.

7. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 1, feature exist In: the micropore cup includes cup plate (6)；It is provided with several indents on the cup plate (6) and is used to place colloidal gold-SPA multiple Close the cup-like trough (7) of object；The sealing element is to be set to a glass plate (6) surface and the opening position for making all cup-like troughs (7) Any one of sealed membrane or micropore lid (8) of sealing.

8. brucella antibody test colloidal gold colloidal gold detection test paper strip in cattle and sheep cream according to claim 7, feature exist In: when the sealing element is micropore lid (8), the micropore lid (8) includes equal with cup-like trough (7) quantity and and cup-like trough (7) one-to-one seal closure；Each seal closure lower end is equipped with the protrusion isometrical with the opening of cup-like trough (7), so that envelope Lid can be sticked in the opening of cup-like trough (7) and seal cup-like trough (7).