KR101894489B1 - Igm semiquantitative diagnostic kit for leptospirosis - Google Patents

Abstract

The present invention relates to a leptospirosis diagnostic kit comprising a first kit and a second kit, and a diagnosis method using the same, wherein the leptospirosis patients can be distinguished and diagnosed not only in the epidemic region but also in the endemic region .

Description

IGM Semi-quantitative Leptospirosis Diagnostic Kit {IGM SEMIQUANTITATIVE DIAGNOSTIC KIT FOR LEPTOSPIROSIS}

The present invention relates to a leptospirosis diagnostic kit comprising a first kit and a second kit, and a diagnostic method for leptospirosis.

Leptospirosis is a common infectious disease of people and animals caused by infection with leptospira interrogans , a type of Spirillum . It is an acute febrile illness with jaundice and renal disease and was named Weil's disease in 1887 and was known as an infectious disease. In 1918, the causative organisms were isolated from patients with Wilhelm's disease and named Spirochaetaicterohaemorrhagiae , and then renamed Leptospira icterohaemorrhagiae . Leptospira belongs to the Leptospiraaceae family. Leptospira genus is classified as Leptospira interrogans and Leptospira biflexa according to the presence or absence of pathogenicity. Leptospira genus is classified as Leptospira interrogans and Leptospira biflexa . The pathogenic Leptospira, Terogenes, has been classified as 30 serogroups and has been reported to contain about 300 serovars, which are known to have different serotypes depending on the region. Leptospira interferon can survive for several weeks if the temperature is maintained at about 20 ° C even if it is present in the tissue of an animal infected with leptospira interferon or through the excretion of animal, It can survive on soil or water and infect humans. Thus, humans can be infected primarily through food or water contaminated with leptospirosis intoxicated or treated with tissue from infected animals or infected animals. It is known that leptospirosis occurs mainly in Europe, the Americas, Australia, Vietnam, Thailand, Malaysia, Taiwan, China and Japan. Serologically proven cases of leptospirosis were reported in Korea in 1942, and in 1984, leptospirosis was isolated from patients with pulmonary hemorrhage. Since then, Korea has been designated as a third-party court infectious disease along with Tsutsugamushi disease, syphilis hemorrhagic fever, and rash fever.

Symptoms of leptospirosis are acute febrile illnesses that are typical of clinical symptoms such as fever, headache, muscle aches, and depression at the onset of the disease. In severe cases, it can cause symptoms of internal organs such as pulmonary hemorrhage, jaundice and kidney failure. Leptospirosis is caused by leptospiroses penetrating through mucous membranes or wounded skin, rapidly spreading throughout the blood vessels, penetrating into tissues, and then injuring the endothelial cells of blood vessels by leptospirosis toxin. It causes symptoms such as long-term bleeding, decreased blood volume, hypotension, etc. However, it is important to find antibiotics such as penicillin at the beginning of the onset, so that they are found within one to two days after the onset and within three to four days at the latest.

As a method for diagnosing leptospirosis, there is the World Health Organization (WHO) standard method, MAT (microscopic agglutination test). Compared to other diagnostic methods, MAT has the advantage of accurate diagnosis. However, since living bacteria are used, maintenance and management are troublesome, expensive equipment is required, and only a skilled expert can test it. Therefore, it is necessary to develop a simple, rapid and accurate method for diagnosing leptospirosis at medical institutions such as public health centers and hospitals. Recently, a rapid kit using immunochromatography assay has been developed. The Rapid Kit is a medical device developed to enable rapid and accurate diagnosis of leptospirosis in 15 minutes with the concept of on-site diagnostic point-of-care testing (POCT). However, such a leptospirosis diagnostic kit is generally developed to be used without distinguishing between an epidemic region and an endemic region of leptospirosis. However, in the case of using the kit in the endemic region of the leptospirosis, the problem is that the antibodies present in the past infected patients or in patients with inapparent infection can be diagnosed as leptospirosis patients have. Therefore, it was necessary to develop a kit that can diagnose leptospirosis by distinguishing between epidemic region and endemic region.

It is an object of the present invention to provide a leptospirosis diagnostic kit and a diagnostic method using the same, and more particularly, to provide a leptospirosis diagnostic kit comprising the first kit and the second kit and a diagnostic method using the same.

However, the problems to be solved by the present invention are not limited to the above-mentioned problems, and other problems not mentioned can be clearly understood by those skilled in the art from the following description.

A first aspect of the present invention is a kit comprising a first kit and a second kit, wherein the first kit comprises 1.2 to 2.0 μg of Leptospira antigens and the second kit comprises 0.6 to 0.9 μg of Leptospira antigens and BSA, and the leptospira antigen is a polysaccharide derived from a lipopolysaccharide of Leptospira.

According to one embodiment, the Ab titer measured by MAT can be determined from 1:50 to 1: 200 using the Leptospirosis diagnostic kit.

According to one embodiment, the BSA may be between 0.1 and 0.4 μg.

According to one embodiment, the first kit further comprises a gold-conjugate-goat-anti-human IgM antibody of OD 3.5 to 4.5, and the second kit comprises a gold-conjugate-goat- An anti-human IgM antibody.

According to one embodiment, the kit may be a strip type using immunochromatography, a device type using immunochromatography or a microplate type.

According to one embodiment, the strip-type kit may include a nitrocellulose membrane, a conjugate pad, an absorbent pad, and a sample pad.

In a second aspect of the present invention, there is provided a method for diagnosing a leptospirosis patient having an Ab titer of 1:50 to 1: 200 measured by a MAT using the above-described leptospirosis diagnostic kit, Lt; RTI ID = 0.0 > IgM < / RTI > antibody.

According to one embodiment, the patient's sample may be serum, plasma or whole blood.

According to one embodiment, the method may further include comparing the detection of the leptospira IgM antibody in the first kit with the detection of the leptospira IgM antibody in the second kit.

According to one embodiment, the step of detecting the antibody uses an antigen-antibody reaction, and the antigen-antibody reaction is detected by enzyme immunoassay (ELISA), radioimmunoassay (RIA), sandwich assay, One or more methods selected from the group consisting of Western blotting, immunoprecipitation, immunohistochemical staining, flow cytometry, fluorescence activated cell sorting (FACS), enzyme substrate staining and antigen-antibody aggregation Can be used.

The leptospirosis diagnostic kit comprising the first kit and the second kit according to the present invention and the diagnostic method using the same can diagnose a leptospirosis patient having an Ab titer of 1:50 to 1: 200 measured by MAT Therefore, it is possible to diagnose leptospirosis patients not only in the epidemic region but also in the endemic region.

FIG. 1 shows a strip-type leptospirosis diagnostic kit according to an embodiment of the present invention.

In the following, embodiments will be described in detail with reference to the accompanying drawings. Like reference symbols in the drawings denote like elements.

Various modifications may be made to the embodiments described below. It is to be understood that the embodiments described below are not intended to limit the embodiments, but include all modifications, equivalents, and alternatives to them.

The terms used in the examples are used only to illustrate specific embodiments and are not intended to limit the embodiments. The singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, the terms "comprises" or "having" and the like refer to the presence of stated features, integers, steps, operations, elements, components, or combinations thereof, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this embodiment belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.

In the following description of the present invention with reference to the accompanying drawings, the same components are denoted by the same reference numerals regardless of the reference numerals, and redundant explanations thereof will be omitted. In the following description of the embodiments, a detailed description of related arts will be omitted if it is determined that the gist of the embodiments may be unnecessarily blurred.

As used herein, the term " diagnosis " means identifying the presence or characteristic of a pathological condition. For the purposes of the present invention, the diagnosis is also to identify leptospirosis.

The term " leptospirosis " as used herein refers to diseases caused by pathogenic strains of Leptospira.

The term epidemic region used in the present invention refers to an area in which a specific epidemic disease occurs intensively at a certain time.

The term endemic region used in the present invention refers to an area in which a specific infectious disease occurs continuously.

The leptospirosis diagnostic kit of the present invention comprises a first kit and a second kit, wherein the first kit comprises 1.2 to 2.0 μg of Leptospira antigens and the second kit comprises 0.6 to 1.0 μg of Leptospira antigens and BSA, and the leptospira antigen is a polysaccharide derived from a lipopolysaccharide of Leptospira.

In order to detect antibodies against leptospira, the leptospirosis diagnostic kit of the present invention utilizes a polysaccharide in which lipids are separated from the lipopolysaccharide (LPS) of the genus Leptospira as an antigen. Lipopolysaccharide (LPS) of leptospirosis is composed of lipid A, core polysaccharide and O-specific side chain. Lipid A and core polysaccharides are linked by 2-keto-3-deoxyoctanoic acid (KDO), and the core polysaccharides are composed of glucose, galactose and N-acetylglucosamine. More specifically, a polysaccharide composed of a genus-specific O-specific side chain and a core polysaccharide from which lipid A has been removed to increase antigenicity of the antigen is used as an antigen (see Example 1).

Since the antigen and the gold-conjugate concentration of the conventional leptospirosis diagnostic kit were intended to detect the presence or absence of the antibody, the amount of the antibody was arbitrarily selected so that only the presence or absence of color due to the antigen-antibody reaction could be discriminated . However, the leptospirosis diagnostic kit of the present invention is a diagnostic kit for detecting the presence or absence of an antibody present in a patient, and for selectively coloring only when a certain amount of antibody amount is dispensed in the diagnostic kit. These diagnostic kits are necessary because the amounts of antibodies present in living organisms may differ depending on the epidemic area and the climate area. It may be difficult to make an accurate diagnosis only with a diagnostic kit that measures the presence or absence of antibodies.

The leptospirosis diagnostic kit of the present invention comprises a first kit and a second kit, wherein the first kit comprises 1.2 to 2.0 μg, preferably 1.4 to 1.8 μg, most preferably 1.6 μg of leptospira antigens, The second kit may comprise from 0.6 to 0.9, preferably 0.8, of leptospirosis antigens. The IgM cut-off was 50 (MAT Ab titer 1: 50) to IgM cut-off was 200 (MAT Ab titer 1: 200) through the antigen-antibody reaction in the first kit and the second kit ) Can be identified. The second kit may also comprise BSA or casein. By including BSA or casein, the degree of antigen-antibody response to leptospira can be controlled.

The leptospirosis diagnostic kit of the present invention is a kit for diagnosing leptospirosis by measuring the level of antibody against leptospira in a biological sample. In order to examine the formation of an antigen-antibody complex, leptospira, which acts as an antigen reacting to an antibody against leptospira, And polysaccharides derived from lipid polysaccharides of fungi.

According to one embodiment, the amount of BSA or casein contained in the second kit of the leptospirosis diagnostic kit of the present invention may be 0.1 to 0.4 μg. When BSA is included, it is preferable that the total amount of BSA and leptospira antigens contained in the second kit is 1 占 퐂. If the amount of BSA is more than 0.4, the reactivity of the second kit may be lower than that of the MAT Ab titer of 50 to 200. If the amount of BSA is less than 0.1, the antigen- There may be a problem that the reactivity of the antibody is increased and thus false positives are caused. (See Example 2-3)

According to one embodiment, the first kit further comprises a gold-conjugate-goat-anti-human IgM antibody of OD 3.5 to 4.5, and the second kit comprises a gold-conjugate-goat- An anti-human IgM antibody. When the gold-conjugate-goat-anti-human IgM antibody of the first kit is contained at an OD of less than 3.5, color development due to an antigen-antibody reaction may not be exhibited. The value of MAT Ab may be positive in less than 50 specimens. When the gold-conjugate-goat-anti-human IgM antibody of the second kit is contained at an OD of less than 1.5, color development due to an antigen-antibody reaction may not be exhibited. There is a problem that the MAT Ab gene, which is an off-value, may result in positive in less than 200 samples (see Example 2-3).

Immunoassays for examining antigen-antibody complex formation include Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, immunofluorescence, immunochromatography, FACS, and the like. However, the immunoassay method is not limited thereto. For example, the immunoassay method may be used for immunoassay, rocket immunoelectrophoresis, tissue immuno staining, immunoprecipitation assay, complement fixation assay, It is not.

The kit for diagnosing leptospirosis of the present invention includes not only the polysaccharide specifically binding to an antibody but also tools and reagents generally used in the field used for immunological analysis. Such tools or reagents include, but are not limited to, suitable carriers, labeling substances capable of producing a detectable signal, solubilizers, cleaning agents, buffers, stabilizers, and the like. The diagnostic kit of the present invention may also be of a type such as, but not limited to, a microplate, a dip-stick device, an immunochromatographic test strip, a spin-split immune assay device, a flow-through device, and the like.

A preferred diagnostic kit is a strip-type diagnostic kit or device-type diagnostic kit using immunochromatography. The diagnosis using the immunochromatography method is carried out in such a manner that an antibody contained in a serum sample of a biological sample reacts with a tracer antibody bound to a colloidal gold particle and then the micropores of the nitrocellulose membrane and a colorant band is formed by binding with a capture antigen immobilized on the inner surface of the micropores while moving through a micropore, thereby enabling positive and negative discrimination by visual inspection. The "marker antibody" refers to an antibody that reacts with antibodies against leptospirosis and is bound to color particles.

According to one embodiment, the anti-leptospirosis diagnostic kit of the present invention can use a color particle binding method for the antigen-antibody complex, and examples of the color particles include colloidal gold particles or color glass or plastic (for example, polystyrene, poly Propylene, latex, etc.) beads can be used. More preferably, gold colloid can be used.

FIG. 1 shows a strip-type leptospirosis diagnostic kit according to an embodiment of the present invention.

According to one embodiment, a leptospirosis diagnostic kit of the present invention comprises a sample pad on which a sample is absorbed; A gold conjugation pad comprising a marker antibody that binds to an antibody in the sample; A test membrane treated with a test line containing the polysaccharide of the present invention and a control line containing a control protein and a pre-test line for removing a nonspecific antibody, ; And a test strip comprising an absorption pad on which a sample of the remaining amount is absorbed. The reaction line includes the polysaccharide of the present invention, and the control line includes a control protein of rabbit anti-goat IgG. In addition, the entire reaction line may contain a gram-negative bacterium having a structure similar to the lipopolysaccharide (LPS) of the present Leptospira, for example, a lipopolysaccharide (LPS) such as E. coli or Pseudomonas at a concentration of 0.5 to 1 mg / ml . The gold binding pad may comprise gold-labeled goat anti-human IgM.

According to one embodiment, when the leptospirosis diagnostic kit of the present invention is a strip-type diagnostic kit using immunochromatography, a method of diagnosing leptospirosis is as follows. First, when the biological sample to be examined is dropped on a sample pad, which is a site where the sample of the diagnostic kit is absorbed, the biological sample reaches the gold conjugation pad by capillary phenomenon, Labeled antibody in a gold binding pad, for example a gold-labeled anti-human IgM goat antibody to form a colloid. At this time, the biological sample to be tested is not immobilized on the gold binding pad but continues to move to the test line on which the polysaccharide antigen of the present invention is immobilized. In the case of a biological sample of a patient infected with leptospirosis, the antibody reacts with the polysaccharide antigen, that is, the antibody against the specific marker antibody in the gold binding pad of the patient binds to the polysaccharide antigen immobilized on the reaction line They are combined to form a red purple band, so that they can be observed with the naked eye. In addition, the marker antibody in the gold binding pad that has not reacted with the antibody in the biological sample reaches the control line and reacts with the control protein to form a red purple band, thereby showing the suitability of the test. As described above, the diagnostic kit of the present invention can rapidly confirm the test results with the naked eye without using special equipment by using the immunochromatography method by the antigen-antibody reaction. In addition, the diagnostic kit improves the accuracy of the diagnosis of leptospirosis by eliminating the possibility of false misrepresentation even though the pretest line does not actually contain an antibody against Leptospira.

According to one embodiment, the biological sample from which an antibody against Leptospira can be detected includes, but is not limited to, blood, serum, plasma, and the like.

Example 1 Preparation of Polysaccharide Antigens Derived from Leptospira

1-1. Leptospira culture

EMJH (Leptospira medium base Ellinghausen McCullough Hohnson Harris) was dissolved in 0.23 g / 100 ml sterilized, and 10 ml of leptospira enrichment EMJH was added to the culture medium. The medium was inoculated with leptospirosis and cultured at 24 ° C for 5 days with shaking.

1-2. Preparation of leptospira antigen

Leptospira cultured for 5 days in EMJH medium was centrifuged at 4080 × g for 20 minutes. The cells were washed three times with 1 × PBS buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO ₄ , and 2 mM KH ₂ PO ₄ ) Lt; / RTI > for 1 hour. Proteinase K (150 / / ml) was added thereto and reacted overnight at 37 캜. Then, EGTA was added to make 2 mM, followed by reaction at 70 ° C for 15 minutes, followed by centrifugation at 4 ° C and 180,000 × g for 2 hours. The precipitate was suspended by adding H ₂ O to produce a lipopolysaccharide (Westpha & Jann, 1965). The purified LPS antigen was treated to 1% acetic acid, heated at 100 ° C for 1 hour, centrifuged at 2980xg for 20 minutes, and lipid A was removed to prepare a polysaccharide antigen. The amount of the prepared antigen was measured using a polysaccharide obtained by treating LPS (Sigma) purified from Escherichia coli in the same manner as above as a control.

Example 2: Production of a diagnostic kit

An NC membrane with a plastic backing was used as a basic skeleton to immobilize an antigen in the middle, a polymer membrane-loaded glass fiber membrane (conjugate pad) was attached to the lower part, and a cellulose membrane absorbing an excessive amount of aqueous solution was placed on both ends Pad), and are connected to each other. The glass fiber membrane located at the bottom can absorb the sample within a short time because of its high absorption power and can prolong the stagnation time before the dissolved polymer moves to the upper immunostrip, thereby inducing an efficient reaction between the analyte and the polymer. A specific antibody (control line) capable of detecting an analyte measurement antigen (a test line or a capture line) and a polymer was spatially separated and immobilized on an intermediate immunostrip and immobilized on the top of the immunostrip The cellulose membrane, which is an absorbent pad, allows excess sample to be absorbed quickly.

2-1. Immobilization of a leptospira antigen (polysaccharide) on nitrocellulose membrane

Immobilization of specific antigens, which serve to capture analytical specific antibodies, was carried out by electrostatic interaction on nitrocellulose (NC) membranes. The control antibody (rabbit anti-goat IgG) was placed at a distance of 1 cm from the bottom of the plastic backed NC membrane strip (4 X 25 mm) using a dispenser at 1 ul / cm As shown in Fig. The dripped membrane was allowed to dry at 37 DEG C for 1 hour.

2-2. Antigen crystals to be used in the pre-test line

When a nonspecific signal due to a polysaccharide occurs, it can be removed by dropping the pretest line in front of the antigen-presenting site to remove the nonspecific antibody. LPS of E. coli, Psudomonasaeruginosa, somewhat similar to Leptospira LPS, was used.

2-3. Antigen concentration and gold-conjugate concentration

Concentrations of antigens and gold-conjugates of the conventional leptospirosis diagnostic kit were used to detect the presence or absence of antibody due to antigen-antibody reaction. However, in addition to measuring the presence or absence of antibodies, it is important to determine the antigens and gold-conjugate concentrations that allow selective color development only when a certain range of antibody amounts are dispensed into the diagnostic kit. This is because, as mentioned above, the amount of antibody present in living organisms may differ depending on the epidemic area and the climate region, and therefore, it is difficult to make an accurate diagnosis by simply detecting the presence or absence of the antibody.

(1) antigen concentration and gold conjugate concentration

Concentrations of the antigen in the kit were adjusted to 0.2, 0.4, and 0.8 μg per kit, and gold-conjugate ODs were set to 1, 2, 3, and 4, respectively. Antibodies used were those with standard IgM, titers 160, 320 and 640, respectively.

Table 1 shows kit responses according to antigen concentration and Gold-conjugate OD.

As can be seen from Table 1, the amounts of detectable antibodies were different depending on the antigen concentration and the OD of the gold-conjugation. Especially, when the gold-conjugation OD value was 2, significant results were obtained depending on the amount of the antigen and antibody .

(2) Second kit antigen concentration

The gold-conjugate OD value in the kit was 2, and the concentrations of the antigens were 0.2, 0.4, and 0.8 μg per kit, respectively. The response of the kit was tested using 10 antibodies with IgM titers of 160, 320, 640 in each kit. The antibodies used are as follows.

- Potency 160: 99-7, 00-224, 01-224, 87-109, 88-123

- Potency 320: 90-226, 95

- Titer 640: 99-4, 00-100, 91-188

Tables 2 to 4 show the kit responses according to the concentration of antigen and antibody.

When the gold-conjugate OD value was fixed to 2 and the degree of color development according to the amount of the antigen and the amount of the antibody was tested, when the amount of the antigen was 0.4 쨉 g or less, the degree of color development was low, There is a problem that it is not easy to distinguish according to the above.

(3) BSA concentration

BSA was added to the loaded antigen in the kit, so that the total amount to be dispensed was 1 占 퐂 / kit. Each kit contained 0.1 μg antigen + 0.9 μg BSA, 0.2 μg antigen + 0.8 μg BSA, 0.4 μg antigen + 0.6 μg BSA, 0.6 μg antigen + 0.4 μg BSA and 0.8 μg antigen + 0.2 μg BSA, respectively. The response of the kit was tested using 10 antibodies with IgM titers of 160, 320, 640 in each kit. The antibodies used are as follows.

- Potency 160: 99-7, 00-224, 01-224, 87-109, 88-123

- Potency 320: 90-226, 95

- Titer 640: 99-4, 00-100, 91-188

Tables 5 to 9 show kit responses according to the concentration of antigen and BSA.

The addition of BSA made it easier to distinguish the degree of color development depending on the amount of antibody. In the case of 0.6 항 antigen + 0.4 B BSA and 0.8 항 antigen + 0.2 B BSA, IgM antibody titer 160 and IgM antibody titer 320 were preferable.

(4) First kit antigen concentration

Twelve kits containing 0.8 μg, 1.6 μg, or 3.2 μg of antigen per kit were prepared and tested three times (four per cycle).

When the concentration of antigen was 1.6 ㎍, the degree of color development according to the reaction in the kit was the best without noise.

2-4. Concentration of sample

The concentration of the human serum used in the kit can also induce a nonspecific reaction when the amount of the antigen or the gold conjugate in the kit is excessive or small. Therefore, if the specimen used in the diagnostic kit is serum or plasma, take 3 μl and dilute it to 300 μl with a diluent (100 μl). If whole blood is 6 μl, take 300 μl of the diluted solution It is preferable to use it by diluting 50 times. Take 300 μl of this diluted specimen with a micropipette and inject it into the specimen injector. Alternatively, 3 μl for serum or plasma and 6 μl for whole blood are taken with a micropipette, so as to be absorbed by the sample pad at the bottom of the specimen injector, and immediately 7 drops of sample diluent are injected into the specimen injector .

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. For example, if the techniques described are performed in a different order than the described methods, and / or if the described components are combined or combined in other ways than the described methods, or are replaced or substituted by other components or equivalents Appropriate results can be achieved.

Therefore, other implementations, other embodiments, and equivalents to the claims are also within the scope of the following claims.

Claims (8)

A leptospirosis diagnostic kit comprising a first kit and a second kit,
Wherein said first kit comprises 1.2 쨉 g to 2.0 쨉 g of leptospira antigens,
Wherein said second kit comprises 0.8 내지 to 0.9 의 of leptospira antigen and 0.1 내지 to 0.2 의 of BSA,
The total amount of leptospira antigens and BSA contained in the second kit was 1 μg,
The leptospira antigen is a polysaccharide derived from a lipopolysaccharide of Leptospira,
The above-mentioned leptospirosis diagnostic kit specifically diagnoses a leptospirosis patient at the time of diagnosis within the epidemic region or the climate region. delete The method according to claim 1,
A leptospirosis diagnostic kit, wherein the Ab titer measured by MAT is 1:50 to 1: 200. delete The method according to claim 1,
The first kit further comprises a gold-conjugate-goat-anti-human IgM antibody of OD 3.5 to 4.5, and
Wherein said second kit further comprises a gold-conjugate-goat-anti-human IgM antibody at an OD of 1.5 to 2.5. To identify leptospirosis according to the epidemic area or the climate area,
In order to provide the information necessary for diagnosing a leptospirosis patient having an Ab titer of 1: 50 to 1: 200 as measured by MAT,
A method for providing information for the diagnosis of leptospirosis, comprising the step of detecting the leptospira IgM antibody in a sample of a patient using the leptospirosis diagnostic kit of claim 1. The method according to claim 6,
Comparing the detection of the leptospira IgM antibody in the first kit with the detection of the leptospira IgM antibody in the second kit. The method according to claim 6,
The step of detecting the antibody uses an antigen-antibody reaction,
The antigen-antibody reaction can be assayed by enzyme immunoassay (ELISA), radioimmunoassay (RIA), sandwich assay, Western blotting, immunoprecipitation, immunohistochemical staining, wherein the method comprises using one or more methods selected from the group consisting of flow cytometry, fluorescence activated cell sorting (FACS), enzyme substrate staining and antigen-antibody aggregation.

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