CN216979096U - Chemiluminescence IgG detection device - Google Patents
Chemiluminescence IgG detection device Download PDFInfo
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- CN216979096U CN216979096U CN202123365802.7U CN202123365802U CN216979096U CN 216979096 U CN216979096 U CN 216979096U CN 202123365802 U CN202123365802 U CN 202123365802U CN 216979096 U CN216979096 U CN 216979096U
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Abstract
The utility model discloses a chemiluminescent IgG detection device, wherein a chip moving module drives a chip reaction tank and a chip in the chip reaction tank to move in an X direction, a reagent extruding device moves to the position right above a required reagent container through the X direction moving module, the reagent container is extruded under the driving of a Z direction moving module to release a reagent in the reagent container into the chip reaction tank, a CCD imager reads a chemiluminescent signal on the chip, 1-10 reaction channels are arranged on the chip, a plurality of microspheres are loaded in each reaction channel, and each microsphere in the same reaction channel is a microsphere coated with different antigens. The chemiluminescent IgG detection device provided by the utility model simultaneously embeds multiple microspheres coated with different types of antigens into one microchip, eliminates the interference of cross reaction among multiple projects on the result, can be used for simultaneously detecting multiple viruses, and has the advantages of short reaction time and small consumption of samples and reagents.
Description
Technical Field
The utility model relates to a chemiluminescent IgG detection device.
Background
Toxoplasmosis, also known as toxoplasmosis, is a zoonosis caused by Toxoplasma gondii, and is most important when cats and felines transmit the disease. After the pregnant women are infected with the toxoplasma gondii, the toxoplasma gondii can be vertically spread to fetuses through the placentas, the development of the fetuses is damaged, the serious mental system development deformity and the intellectual disability of the fetuses are caused, the population quality is reduced, and even the fetuses die. The TOX-IgM antibodies appeared earlier than the TOX-IgG antibodies, which declined faster after the acute phase, and the TOX-IgG antibodies may be present for a long time. Therefore, the determination of the TOX-IgM antibody is suitable for early diagnosis, and the positive TOX-IgG antibody indicates that the Toxoplasma gondii is infected once and is mostly chronic infection.
Rubella virus belongs to the genus rubella of the family enveloped viruses, and comprises a single-stranded, plus-stranded RNA, and three structural proteins E1, E2 and C, wherein glycoproteins E1 and E2 are located on the envelope in heterodimers, and protein C and RNA form the nucleocapsid. The patient with rubella is the source of the rubella. Humans are the only host for rubella virus, which is transmitted primarily through respiratory droplets. After infection of pregnant women, the fetuses can be infected through the placenta. Clinically, fever, retroauricular occipital and retrocervical lymphadenopathy and maculopapular rash are the main manifestations, and a large number of patients present with recessive infection. Pregnant women infected in the early gestation period (the first 3 months) are easy to cause congenital rubella syndrome such as stillbirth, abortion or fetal deformity. Rubella IgG antibodies (RUB-IgG) are usually present at the time of rash and will remain after the rash.
Cytomegalovirus (CMV) is a herpesvirus DNA Virus with a typical herpesvirus morphology. The distribution is wide, people have wide susceptibility to the disease, the transmission ways are various, and the disease belongs to sexually transmitted diseases. The infection is mainly asymptomatic subclinical infection, and CMV infection can cause infections of various systems mainly including diseases of the genitourinary system, the central nervous system and the liver, from mild asymptomatic infection to serious defect or death. After the pregnant women are infected with cytomegalovirus, the virus can infect the fetus through the placenta, causing congenital infection, abortion and dead fetus of the fetus. CMV-IgG appeared 4-6 weeks after viral infection and continued for life.
Herpes Simplex Virus (HSV) is a viral pathogen that seriously harms human health, causes skin and venereal diseases, and can cause infections such as gingivitis, keratoconjunctivitis, encephalitis, and infections of the reproductive system and neonate. Humans are the only host of human HSV, which can be present in the vesicle fluid, saliva and feces of patients, convalescent or healthy carriers, disseminated in vivo via blood or neural pathways, and harbor some form of post-infection HSV in locally infected ganglion cells. Viruses are divided into two serotypes, type I and type II. Among them, HSV-I type infection has low recurrence rate and long attack time interval. After a patient is infected with HSV, antibodies aiming at the HSV appear about 1 week, the peak is reached in 3-4 weeks, the antibody can be continued for years, and the antibody level is correspondingly reduced after the treatment. HSV II is transmitted mainly through sexual life, infects genitals and parts below the waist, and is typically clinically manifested as erythema, papules, etc. of the external genitalia skin mucosa. Infection in pregnant women can cause fetal abortion, congenital malformation, or developmental disorders, lethal or permanent nervous system damage in newborns. HSV II-IgG appears 4-6 weeks after viral infection and usually persists for life.
The current main detection methods for the disease detection comprise: enzyme linked immunosorbent assay, colloidal gold assay, and chemiluminescence assay. The chemiluminescence method has higher sensitivity and specificity, and the detection accuracy is higher than that of an enzyme-linked immunosorbent assay and a colloidal gold assay, so that the chemiluminescence method is the most adopted detection method. Most of the existing kits are single detection kits, only one type of microspheres with antigens are arranged in each chip, 5 diseases are required to be detected during pre-pregnancy detection, the detection time is very long, the detection result cannot be given in time, and the existing detection device cannot meet the requirement of simultaneous detection.
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide a multiple micro-ball type chemiluminescence IgG detection device which can be used for simultaneously detecting multiple virus antibodies.
The technical scheme adopted by the utility model is as follows: the utility model provides a chemiluminescence IgG detection device, includes chip, chip reaction tank, a plurality of reagent container, CCD imager, chip removal module, X to removing module, Z to removing module, reagent extrusion device, a plurality of reagent containers can extrude the pressure deformation, all fix on the support, the rigidity, the chip is placed in the chip reaction tank, chip removal module drives chip reaction tank and the chip in it and carries out the removal of X to, reagent extrusion device removes to required reagent container directly over through X to removing the module, extrudees the reagent container in order to carry out the release of its interior reagent to the chip reaction tank under the drive of Z to removing the module, the CCD imager reads the chemiluminescence signal on the chip, its characterized in that: the chip is provided with 1-10 reaction channels, each reaction channel is loaded with a plurality of microspheres, and each microsphere in the same reaction channel is a microsphere coated with different antigens.
Preferably, the chip moving module comprises a screw motor, a slider and a fixing plate, the chip reaction tank is fixed on the fixing plate of the chip moving module, the fixing plate is fixed on the slider of the chip moving module, and the screw motor and the slider on the chip moving module are matched to drive the chip reaction tank and the chip therein to move in the X direction together.
Preferably, the X-direction moving module comprises a fixed block, an X-direction moving module sliding block and an X-direction moving module lead screw motor, the Z-direction moving module comprises a Z-direction moving module sliding block and a Z-direction moving module lead screw motor, the reagent extruding device is fixed on the Z-direction moving module sliding block, and the Z-direction moving module drives the Z-direction moving module sliding block through the Z-direction moving module lead screw motor so as to drive the reagent extruding device on the Z-direction moving module sliding block to realize Z-direction movement; the Z-direction moving module is fixed on a fixed block of the X-direction moving module, the fixed block is fixed on an X-direction moving module sliding block, and the X-direction moving module drives the X-direction moving module sliding block and the fixed block fixed on the X-direction moving module sliding block through an X-direction moving module screw rod motor on the X-direction moving module to realize that the Z-direction moving module and a reagent extruding device fixed on the Z-direction moving module move together in the X direction.
Preferably, the chip moving module moves the chip to a position right below the CCD imager, so that the CCD imager reads the chemiluminescent signal on the chip.
Preferably, each reaction channel of the chip is loaded with 5 microspheres, and the 5 microspheres are microspheres coated with toxoplasma antigen, microspheres coated with rubella virus antigen, microspheres coated with cytomegalovirus antigen, microspheres coated with herpes simplex virus type 1 antigen and microspheres coated with herpes simplex virus type 2 antigen.
Preferably, the chip is made of polyvinyl chloride, polystyrene or polyacrylamide.
Preferably, the microspheres are made of polyvinyl chloride, polystyrene or polyacrylamide.
Preferably, the microspheres coated with different antigens are arranged in series in the reaction channel, and the arrangement shape is a straight line, a curve, a Z shape or an L shape.
Preferably, the reagent container comprises a washing liquid container, an enzyme labeling reagent container, a sample diluent container and two excitation liquid containers.
The chemiluminescence IgG detection device provided by the utility model simultaneously embeds a plurality of microspheres coated with different types of antigens into a microchip, eliminates the interference of cross reaction among a plurality of projects on the result, can be used for simultaneously detecting a plurality of virus IgG, and has the advantages of short reaction time and small consumption of samples and reagents. In addition to the simultaneous detection of 5 viruses, i.e., TOX, RUB, CMV, HSV-I, HSV-II, it should be particularly noted that the present invention is not limited to the detection of 5 viruses, i.e., TOX, RUB, CMV, HSV-I, HSV-II, and the device of the present invention can also be used for the simultaneous detection of IgA, IgE, or IgD antibodies of more than 3 other viruses, and only different antigens need to be coated on the microspheres.
Drawings
FIG. 1 is a schematic view of the structure of a chemiluminescent IgG detection device.
FIG. 2 is a schematic diagram of a mobile module of a chemiluminescent IgG detection device
FIG. 3 is a schematic diagram of a chip structure. Coating microspheres from left to right: toxoplasma antigen, rubella virus antigen, cytomegalovirus antigen, herpes simplex virus type I antigen, herpes simplex virus type II antigen.
FIG. 4 shows the luminescence of microspheres in a chip.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Example 1
As shown in fig. 1 and 2, a chemiluminescent IgG detection device comprises a chip reaction tank 1, a chip 2, a washing solution container 3, an enzyme-labeled reagent container 4, a sample diluent container 5, a support 6, excitation solution containers 8 and 9, a CCD imager 7, a chip moving module 10, an X-direction moving module 11, a Z-direction moving module 12, and a reagent squeezing device 13. The washing liquid container 3, the enzyme-labeled reagent container 4, the sample diluent container 5 and the excitation liquid containers 8 and 9 are all fixed on the bracket 6 and are fixed in position. The chip 2 is placed in the chip reaction tank 1, the chip reaction tank 1 is fixed on a fixing plate 14 of the chip moving module 10, the fixing plate 14 is fixed on a slide block 16 of the chip moving module 10, and the chip reaction tank 1 and the chip 2 therein can be driven to move in the X direction together through the matching of a screw rod motor 17 on the chip moving module 10 and the slide block 16, so that a liquid inlet on the chip moves to the right below a corresponding reagent container to release a corresponding reagent. The reagent extruding device 13 is fixed on a slide block of the Z-direction moving module 12, and the Z-direction moving module 12 drives the slide block through a screw rod motor thereof so as to drive the reagent extruding device 13 thereon to realize the Z-direction movement. The Z-direction moving module 12 is fixed on a fixing block 15 of the X-direction moving module 11, the fixing block 15 is fixed on a sliding block of the X-direction moving module 11, the X-direction moving module 11 drives the sliding block and the fixing block 15 fixed on the sliding block through a screw rod motor on the X-direction moving module to realize that the Z-direction moving module 12 and a reagent extruding device 13 fixed on the sliding block move together in the X-direction, and the reagent extruding device 13 can be accurately positioned right above a corresponding reagent container through the mutual matching of the X-direction moving module and the Z-direction moving module and extrude the reagent container to release the reagent into a chip reaction tank. After the reagent reaction is completed, the chip moving module 10 moves the chip to a position right below the CCD imager 7, so that the acquisition area of the CCD imager 7 can completely cover the light-emitting area. The CCD imager 7 can read the chemiluminescence signal on the chip after the reagent reaction is completed. The chip 2 is provided with 1 reaction channel, each reaction channel is loaded with 5 microspheres, and the 5 microspheres are microspheres coated with toxoplasma antigen, microspheres coated with rubella virus antigen, microspheres coated with cytomegalovirus antigen, microspheres coated with herpes simplex virus 1 antigen and microspheres coated with herpes simplex virus 2 antigen.
The microsphere and the chip loading frame are both made of polyvinyl chloride materials.
Polystyrene or polyacrylamide materials are also possible.
A plurality of reaction channels can also be arranged on the chip and used for detecting a plurality of samples simultaneously.
The chip reaction tank can move, and when imaging, the chip reaction tank moves into the imaging device for imaging.
Or the chip reaction tank is fixed, the imaging device can move, and the imaging device moves to the chip reaction tank for imaging during imaging.
Example 2
1. The microspheres are made of polyvinyl chloride.
2. Microspheres were treated 2 times with 5 min of washing each time using ph7.4 buffer.
3. Adding toxoplasma antigen into 5ml of phosphate buffer solution with pH7.4 according to the concentration of 1ug/ml to prepare coating solution, putting 20 microspheres into 10ml of coating solution, treating for 16-24 hours at 2-8 ℃, washing for 6 times by using buffer solution with pH6.5-pH7.5, and treating for 16 hours at 30 ℃.
4. Adding rubella virus antigen into 5ml of phosphate buffer solution with pH7.4 according to the concentration of 1ug/ml to prepare coating solution, putting 20 microspheres into 10ml of coating solution, treating for 16-24 hours at 2-8 ℃, washing for 6 times by using buffer solution with pH6.5-pH7.5, and treating for 16 hours at 30 ℃.
5. Adding cytomegalovirus antigen into 5ml of phosphate buffer solution with pH value of 7.4 according to the concentration of 2ug/ml to prepare coating solution, placing 20 microspheres into 10ml of coating solution, treating for 16-24 hours at 2-8 ℃, washing for 6 times by using buffer solution with pH value of 6.5-7.5, and placing the microspheres coated with the cytomegalovirus antigen at 30 ℃ for 16 hours.
6. Adding 5ml of carbonic acid buffer solution with pH9.6 into the single-purity herpes virus I antigen according to the concentration of 2ug/ml to prepare coating solution, putting 20 microspheres into 10ml of coating solution, treating for 16-24 hours at 2-8 ℃, washing for 5 times by using buffer solution with pH6.5-pH7.5, washing for 1 time by using buffer solution with pH6.5-pH7.5, and treating for 16 hours at 30 ℃.
7. Adding 5ml of carbonic acid buffer solution with pH9.6 into the single-purity herpes virus II antigen according to the concentration of 2ug/ml to prepare coating solution, putting 20 microspheres into 10ml of the coating solution, treating for 16-24 hours at 2-8 ℃, washing for 6 times by using the buffer solution with pH6.5-pH7.5, and treating the microspheres coated with the single-purity herpes virus II antigen for 16 hours at 30 ℃.
8. Five different types of antigens coated are fixed on a polyacrylamide chip in sequence, and the distance between the microspheres can be 1cm, 2cm, 3cm and 4 cm. The completed chip is shown in fig. 3.
Example 3
Use of a chip
1. Placing the chip in a chip reaction tank;
2. adding 50ul of sample to be detected into a chip reaction tank;
3. the sample diluent in the sample diluent container was added to 50ul to the chip reaction chamber.
4. The temperature of the chip reaction chamber area was controlled at 37 ℃ for 10 minutes.
5. And adding the washing liquid in the washing liquid container into the reaction tank, and cleaning the reaction tank.
6. Adding the enzyme-labeled reagent in the enzyme-labeled reagent container into the reaction tank.
7. The temperature of the chip reaction chamber area was controlled at 37 ℃ for 10 minutes.
8. And automatically adding the washing liquid in the washing liquid container into the reaction tank, and cleaning the reaction tank.
9. The exciting liquid A in the exciting liquid container 8 is added to the reaction tank.
10. The exciting liquid B in the exciting liquid container 9 is added to the reaction tank.
And 11, immediately collecting the chemiluminescence signals on the microspheres by the CCD imager, sending the chemiluminescence signals to analysis software for analysis, generating specific numerical values on a software interface, and automatically displaying the positive and negative values and the specific numerical values of the five indexes by the system. As shown in fig. 4 and table 1, the samples were positive for rubella virus IgG, positive for cytomegalovirus IgG, and negative for the remaining three. Wherein the rubella virus IgG value is 800, the cytomegalovirus IgG value is 1600, and the other three items are 0.
Wherein
Enzyme labeling reagent: HRP-labeled anti-human IgG is stored in a buffer solution containing protective protein and contains a preservative;
sample diluent: the phosphate buffer solution specifically comprises: 137mmol/L NaCl, 2.7mmol/L KCl, 44.3 mmol/L Na2HPO44, 4mmol/L KH2PO41.4mmol/L;
washing liquid: phosphate buffer between Ph6.5-7.5;
reaction substrate a: 0.01g/L-10g/L of luminol salt solution;
reaction substrate B: 0.01g/L to 10g/L of hydrogen peroxide solution.
TABLE 1 results of sample analysis
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. The utility model provides a chemiluminescence IgG detection device, includes chip, chip reaction tank, a plurality of reagent container, CCD imager, chip removal module, X to removing module, Z to removing module, reagent extrusion device, a plurality of reagent containers can extrude the pressure deformation, all fix on the support, the rigidity, the chip is placed in the chip reaction tank, chip removal module drives chip reaction tank and the chip in it and carries out the removal of X to, reagent extrusion device removes to required reagent container directly over through X to removing the module, extrudees the reagent container in order to carry out the release of its interior reagent to the chip reaction tank under the drive of Z to removing the module, the CCD imager reads the chemiluminescence signal on the chip, its characterized in that: the chip is provided with 1-10 reaction channels, each reaction channel is loaded with a plurality of microspheres, and each microsphere in the same reaction channel is a microsphere coated with different antigens.
2. The chemiluminescent IgG detection device of claim 1, wherein: the chip moving module comprises a screw rod motor, a sliding block and a fixing plate, the chip reaction tank is fixed on the fixing plate of the chip moving module, the fixing plate is fixed on the sliding block of the chip moving module, and the screw rod motor and the sliding block on the chip moving module are matched to drive the chip reaction tank and a chip therein to move in the X direction together.
3. The chemiluminescent IgG detection device of claim 2, wherein: the X-direction moving module comprises a fixed block, an X-direction moving module sliding block and an X-direction moving module lead screw motor, the Z-direction moving module comprises a Z-direction moving module sliding block and a Z-direction moving module lead screw motor, the reagent extruding device is fixed on the Z-direction moving module sliding block, and the Z-direction moving module drives the Z-direction moving module sliding block through the Z-direction moving module lead screw motor so as to drive the reagent extruding device on the Z-direction moving module sliding block to realize Z-direction movement; the Z-direction moving module is fixed on a fixed block of the X-direction moving module, the fixed block is fixed on an X-direction moving module sliding block, and the X-direction moving module drives the X-direction moving module sliding block and the fixed block fixed on the X-direction moving module sliding block through an X-direction moving module screw rod motor on the X-direction moving module to realize that the Z-direction moving module and a reagent extruding device fixed on the Z-direction moving module move together in the X direction.
4. The chemiluminescent IgG detection device according to claim 1, wherein: the chip moving module moves the chip to be right below the CCD imager so that the CCD imager reads the chemiluminescence signal on the chip.
5. The chemiluminescent IgG detection device according to claim 1, wherein: each reaction channel of the chip is loaded with 5 microspheres, and the 5 microspheres are microspheres coated with toxoplasma antigen, microspheres coated with rubella virus antigen, microspheres coated with cytomegalovirus antigen, microspheres coated with herpes simplex virus 1 antigen and microspheres coated with herpes simplex virus 2 antigen.
6. The chemiluminescent IgG detection device of any of claims 1-5, wherein: the chip is made of polyvinyl chloride, polystyrene or polyacrylamide.
7. The chemiluminescent IgG detection device according to claim 6, wherein: the microspheres are made of polyvinyl chloride, polystyrene or polyacrylamide.
8. The chemiluminescent IgG detection device of claim 1 or 5, wherein: the microspheres coated with different antigens are arranged in series in the reaction channel, and the arrangement shape is a straight line, a curve, a Z shape or an L shape.
9. The chemiluminescent IgG detection device according to any of claims 1 to 5, wherein: the reagent container comprises a washing liquid container, an enzyme-labeled reagent container, a sample diluent container and two exciting liquid containers.
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| CN202123365802.7U CN216979096U (en) | 2021-12-29 | 2021-12-29 | Chemiluminescence IgG detection device |
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