Method and reagent group for diagnosing canine pancreatitis and application of acyl carnitine in diagnosis of canine pancreatitis

Abstract

The invention provides a method for diagnosing canine pancreatitis. First, a specimen of a canine animal is provided. Then, a concentration of a biomarker of the specimen is detected, if the concentration is higher than a predetermined value, it is determined that the canine has a high risk of pancreatitis, the biomarker being medium-chain acyl carnitine, long-chain acyl carnitine, or a combination thereof, where the medium-chain acyl group has a chain length of 6 to 12 carbon atoms, the long-chain acyl group has a chain length of 6 to 12 carbon atoms, and the long-chain acyl group has a chain length of 6 to 12 carbon atoms. And the long-chain acyl refers to the chain length of more than 12 carbon atoms. The invention further provides a reagent group for diagnosing the canine pancreatitis and application of the acyl carnitine to diagnosis of the canine pancreatitis.

Description

Technical Field

[0001] The present invention relates to a method for diagnosing pancreatitis in canines and a reagent set for diagnosing pancreatitis in canines, and more particularly to a method or reagent set for diagnosing pancreatitis in canines by detecting specific biomarkers. Background Art

[0002] Acute pancreatitis is caused by the abnormal activation of trypsinogen in the pancreas, resulting in the ineffective operation of trypsin inhibitor in pancreatic acinar cells and triggering autodigestion, leading to inflammation. It is one of the most severe diseases of the canine digestive tract. Mild pancreatitis is usually accompanied by clinical symptoms such as mild to moderate anorexia, abdominal pain, and vomiting. Appropriate symptomatic treatment usually leads to complete recovery. However, severe acute pancreatitis is characterized by obvious pancreatic necrosis, usually accompanied by severe systemic inflammatory response and multiple organ dysfunction, with a fatality rate approaching 50%.

[0003] Currently, the clinical diagnosis of canine acute pancreatitis is still difficult, especially in the judgment of severity. The currently commonly used diagnostic marker is canine pancreatic-specific lipase (cPL), but its specificity and sensitivity still need improvement, and it cannot clearly distinguish the severity of acute pancreatitis. In actual diagnosis, the medical history, clinical symptoms, and abdominal ultrasound results still need to be considered. Summary of the Invention

[0004] The present invention thus provides a method for diagnosing or detecting pancreatitis in canines and a reagent set for detecting pancreatitis in canines or for its detection. By detecting the concentration of specific biomarkers in a sample, it can quickly assist in diagnosing the possibility of pancreatitis in canines or in differentiating the severity of acute pancreatitis in canines.

[0005] According to an embodiment of the present invention, a method for diagnosing pancreatitis in canines is provided. First, a sample of a canine is provided. Then, the concentration of a biomarker in the sample is detected. If the concentration is higher than a predetermined value, it is determined that the canine has a high risk of pancreatitis, and the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof. The medium-chain acyl group refers to a carbon chain with 6 - 12 carbon atoms, and the long-chain acyl group refers to a carbon chain with more than 12 carbon atoms.

[0006] According to another embodiment of the present invention, the present invention provides a reagent kit for diagnosing canine pancreatitis. The reagent kit includes a detection agent, which, after being mixed with a sample of a canine, can be used to detect the concentration of a biomarker in the sample. The biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, where the medium-chain acyl refers to a carbon chain length of 6-12 carbon atoms, and the long-chain acyl refers to a carbon chain length greater than 12 carbon atoms.

[0007] According to still another embodiment of the present invention, the present invention provides the use of medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof as a biomarker for diagnosing canine pancreatitis, where the medium-chain acyl refers to a carbon chain length of 6-12 carbon atoms, and the long-chain acyl refers to a carbon chain length greater than 12 carbon atoms.

[0008] In one embodiment of the present invention, the medium-chain acylcarnitine referred to by the aforementioned biomarker refers to L-decanoyl carnitine (Decanoyl-L-carnitine, C10), L-dodecanoyl carnitine (Dodecanoyl-L-carnitine, C12), or L-dodecenoyl carnitine (Dodecenoyl-L-carnitine, C12:1), and the sample includes urine or blood.

[0009] In another embodiment of the present invention, the long-chain carnitine referred to by the aforementioned biomarker refers to L-tetradecadienoyl carnitine (Tetradecadienoyl-L-carnitine, C14:2), and the sample includes urine or blood.

[0010] By detecting the specific biomarker provided by the present invention, namely medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, it can be used to assist in diagnosing whether a canine has pancreatitis. Once it is found that a canine has a high possibility of pancreatitis, treatment can be carried out immediately, which can greatly improve its survival rate. Brief Description of the Drawings

[0011] Figure 1 It is a bar graph showing the relative contents of 15 groups of acylcarnitines with different carbon lengths in the sera of healthy dogs and dogs with pancreatitis analyzed by the present invention.

[0012] Figure 2 It is a bar graph showing the relative contents of 13 groups of acylcarnitines with different carbon lengths in the urine of healthy dogs and dogs with pancreatitis analyzed by the present invention. Detailed Description of the Embodiments

[0013] To enable those skilled in the art to further understand the present invention, the preferred embodiments of the present invention will be listed in the following description and will be described in detail in conjunction with the drawings to explain the composition and the intended effects of the present invention.

[0014] The main causes of acute pancreatitis are not yet clear. Common speculated causes are: high-fat diet, improper diet, obesity, drugs and toxins, endocrine diseases, and genetics. Taking dogs as an example, pancreatitis mostly occurs in middle-aged and elderly dogs, and a small number are in dog breeds prone to it, such as: Miniature Schnauzer. Acylcarnitine is one of the intermediate products of fatty acid metabolism, which is produced by the esterification of fatty acids and L-carnitine. Since the regulation of fatty acid metabolism will change the content of acylcarnitine, acylcarnitine can reflect diseases related to fatty acid oxidation disorders and metabolic disorders. If the β-oxidation pathway of fatty acids is affected, the acylcarnitine will increase significantly. As a product of the fatty acid metabolism pathway, acylcarnitine exists in body fluids such as serum and urine. At the same time, acylcarnitine has the function of activating the signal transduction related to the inflammatory response. In the case of high content, it can directly affect the normal physiological functions of cells.

[0015] To solve the problem in the prior art of lacking good biomarkers for detecting pancreatitis in canines, the present invention provides a method for diagnosing pancreatitis in canines. First, a specimen of a canine is provided. In an embodiment of the present invention, the canine may be, for example: a dog, a wolf, a jackal, or a fox, etc., but is not limited thereto. In a preferred embodiment, the canine refers to a dog. On the other hand, the specimen can be obtained from the canine in various ways so that the specimen is separated from the canine and obtained as a subject for detection independent of the canine. In an embodiment, the specimen can be various tissue fluids or body fluids. In a preferred embodiment, the specimen refers to blood, especially serum, which can be obtained by, for example, blood sampling. In another preferred embodiment, the specimen refers to urine, which can be obtained by the canine's own urination, or by other invasive methods such as setting a catheter or bladder puncture.

[0016] Then, a concentration of a biomarker of the specimen is detected. If the concentration is higher than a predetermined value, it is determined that the canine has a high risk of pancreatitis. In an embodiment of the present invention, the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, where the medium-chain acyl refers to a carbon chain length of 6-12 carbon atoms, and the long-chain acyl refers to a carbon chain length greater than 12 carbon atoms. In a preferred embodiment of the present invention, the biomarker refers to medium-chain acylcarnitine. If the specimen detects both blood and urine, the biomarker is preferably Decanoyl-L-carnitine (C10), Dodecanoyl-L-carnitine (C12), or Dodecenoyl-L-carnitine (C12:1). In another optimal embodiment, the biomarker refers to long-chain acylcarnitine. If the specimen detects both blood and urine, the biomarker is preferably Tetradecadienoyl-L-carnitine (C14:2).

[0017] On the other hand, the detection of the concentration of the biomarker mentioned above is carried out by using liquid chromatography-mass spectrometry (LC / MS). In this case, if the concentration is higher than a predetermined value, it can be considered that the canine has a high risk of pancreatitis. The aforementioned predetermined value refers to the concentration of the biomarker in the sample of a healthy canine. Theoretically, for the same canine, this predetermined value should be fixed (predetermined). This predetermined value is obtained through the following method: First, a healthy canine is provided, which is of the same species as the diseased canine to be tested (for example, both are domestic dogs). Then, a healthy sample of the healthy canine is collected, such as serum in blood or urine, and the concentration of the same biomarker (for example, lauroyl carnitine) in the healthy sample is measured by an instrument. Finally, the concentration of the biomarker in the healthy sample is compared with the concentration of the biomarker in the sample to be tested. If the concentration of the biomarker in the sample to be tested is greater than the concentration of the biomarker in the healthy sample (that is, greater than the predetermined value) by a certain proportion, it is considered that the canine to be tested has a high risk of pancreatitis. In an embodiment of the present invention, when the sample is serum and the concentration of the biomarker in the sample to be tested is more than 2 times the concentration of the biomarker in the healthy sample, it is considered that the canine to be tested has a high risk of pancreatitis. In an embodiment of the present invention, when the sample is urine and the concentration of the biomarker in the sample to be tested is more than 20 times the concentration of the biomarker in the healthy sample, it is considered that the canine to be tested has a high risk of pancreatitis. By comparing with the predetermined value, it can be determined whether the canine has a high risk of pancreatitis. Moreover, the so-called having a high risk of pancreatitis in the present invention means that the canine can be diagnosed as having pancreatitis according to traditional diagnostic methods, for example: clinical symptoms include vomiting, diarrhea, loss of appetite, abdominal pain (at least two of the symptoms are met); the value of canine-specific lipase (cPL): >200 μg / L, or whole abdominal ultrasound examination: hypoechoic pancreatic parenchyma, pancreatic thickening, blurred pancreatic margin, high echo adipose tissue around, and other abnormalities such as pancreatic duct / bile duct dilation and ascites, etc., but not limited thereto, and also includes other diagnostic indicators sufficient to represent having pancreatitis.

[0018] In another embodiment of the present invention, the present invention further provides a reagent kit for diagnosing canine pancreatitis. The reagent kit includes a detection reagent. After the detection reagent is mixed with a sample of a canine, it can be used to detect the concentration of a biomarker, which refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, where the medium-chain acyl refers to a carbon chain length of 6-12 carbon atoms, and the long-chain acyl refers to a carbon chain length greater than 12 carbon atoms. The detection reagent includes an internal standard required to detect the concentration of medium / long-chain acylcarnitine. Measuring the concentration of medium / long-chain acylcarnitine can be various methods known in the art, such as detecting the concentration of its internal biomarker by liquid chromatography-mass spectrometry or high-performance liquid chromatography. In one embodiment, the reagent kit of the present invention can also use enzyme immunoassay (ELISA), affinity chromatography column purification method, or modified tube binding assay, etc., to diagnose medium / long-chain acylcarnitine in a non-invasive manner. For example, the present invention uses the ELISA method to detect canine pancreatitis. First, the medium / long-chain acylcarnitine of the present invention is linked to a labeling substance, including, but not limited to, biotin, His tag, fluorescent substance (Cy3 or Cy5), or Dig (Digoxigenin), etc., and by detecting the absorbance value or fluorescence intensity, the amount of medium / long-chain acylcarnitine in the sample is analyzed to determine whether it is at risk of canine pancreatitis. The relevant implementation manners of this embodiment that are the same as those of the foregoing embodiments, such as canines, samples, biomarkers, etc., will not be elaborated herein again.

[0019] In yet another embodiment of the present invention, there is provided the use of medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof for diagnosing pancreatitis in canines, where the medium-chain acyl refers to a carbon chain length of 6-12 carbon atoms, and the long-chain acyl refers to a carbon chain length greater than 12 carbon atoms. The relevant implementation manners of this embodiment that are the same as those of the foregoing embodiments, such as canines, samples, biomarkers, etc., will not be elaborated herein again.

[0020] To more clearly describe the implementation manners and effects of the present invention, the following will give examples and related experiments and illustrate their detailed steps.

[0021] Example

[0022] The experimental procedure of this example is as follows:

[0023] (I) This study was reviewed and approved by the Institutional Animal Care and Use Committee of National Chiayi University in Taiwan, China, and the approval number is 111026.

[0024] (II) Case collection

[0025] (1) Inclusion criteria for diseased canines

[0026] A. Clinical symptoms: including vomiting, diarrhea, loss of appetite, abdominal pain (at least two of the symptoms must be met).

[0027] B. Canine pancreatic lipase (cPL) value: > 200 μg / L

[0028] C. Whole abdominal ultrasound examination: hypoechoic pancreatic parenchyma, pancreatic thickening, blurred pancreatic margin, high echo adipose tissue around, and other abnormalities such as pancreatic / biliary duct dilation and ascites

[0029] (2) Exclusion criteria for diseased dogs

[0030] A. History of primary gastrointestinal, hepatobiliary, and urinary system diseases

[0031] B. No obvious abnormalities in pancreatic ultrasound, and the pancreatic image cannot be clearly observed by ultrasound

[0032] C. Clinical symptoms lasting for more than 7 days

[0033] (3) Inclusion criteria for healthy dogs

[0034] No abnormal clinical symptoms, normal blood test, abdominal ultrasound, and cPL value

[0035] (III) Blood and urine collection

[0036] (1) Healthy dogs: Blood is collected from the cephalic vein at the health check clinic, and urine is collected by natural urination, once for each.

[0037] (2) Diseased dogs: Blood is collected from the cephalic vein at the clinic. If natural urination is not possible, urine is collected by bladder puncture, and sedation (Alfaxalone, 0.3 mL / kg) is used if necessary.

[0038] (IV) Sample testing

[0039] Metabolite and lipid analysis of blood and urine was performed using an Agilent 1290 UPLC coupled with a 6540-QTOF mass spectrometer (Agilent 1290 UHPLC coupled with 6540-QTOF (UHPLC-QTOF) (Agilent Technologies, Santa Clara, CA)) and analyzed according to the published method [1]. To control the quality of the analysis, blank samples and pooled analytical quality control samples were inserted at the beginning of each batch and every five samples for analytical calibration to ensure the analysis quality. At the same time, at the beginning of each analysis, the performance of the instrument was tested with a mixture of 40 synthetic standards. Each sample was analyzed in triplicate, and the total ion chromatogram was manually inspected. The analytical data were used to detect spectral peaks using MZmine 3 [2] and the ADAP algorithm [3]. The spectral peaks were confirmed by matching them against a self-built metabolite database [4]. The potential metabolite features selected by matching were preprocessed to remove those with >50% missing values, and the remaining features were taken as half of the smallest positive value in the original data. Before performing statistical analysis with MetaboAnalyst, the preprocessed data were normalized by the sum of the total peak areas, log-transformed, and auto-scaled, and then analyzed using MetaboAnalyst 5.0 [5]. The methods cited in this paragraph can be referred to the following literature:

[0040] 1. Liu CT, Raghu R, Lin SH, Wang SY, Kuo CH, Tseng Y, Sheen LY. Metabolomics of Ginger Essential Oil against Alcoholic Fatty Liver in Mice. J. Agric. Food Chem. 2013;61:11231–11240.

[0041] 2. Schmid, R., Heuckeroth, S., Korf, A. et al. “Integrative analysis of multimodal mass spectrometry data in MZmine 3.” Nature Biotechnology (2023); https: / / mzmine.github.io / .

[0042] 3.https: / / mzmine.github.io / mzmine_documentation / module_docs / lc-ms_featdet / featdet_adap_chromatogram_builder / adap-chromatogram-builder.html

[0043] 4.In-house database: the Taiwan University MetaCore Metabolomics Chemical Standard Library.

[0044] 5.https: / / www.metaboanalyst.ca / docs / About.xhtml

[0045] Experiment 1: Detection of Canine Serum

[0046] Compare the sera of healthy canines and canines with pancreatitis, and detect the contents of acylcarnitines with fifteen different carbon lengths in the sera respectively. Please refer to Figure 1, a bar graph showing the relative contents of fifteen groups of acylcarnitines with different carbon chain lengths in the sera of healthy dogs and dogs with pancreatitis according to the present invention. The fifteen groups of acylcarnitines with carbon chain lengths are: L-Hexanoyl L-carnitine (C6), L-Octenoyl-L-carnitine (C8:1), L-Octanoyl-L-carnitine (C8), L-Decanoyl-L-carnitine (C10), L-Dodecenoyl L-carnitine (C12:1), L-Dodecanoyl-L-carnitine (C12), L-Tetradecadienoyl-L-carnitine (C14:2), L-Tetradecenoyl-L-carnitine (C14:1), L-Tetradecanoyl-L-carnitine (C14), L-Hexadecenoyl-L-carnitine (C16:1), L-Hexadecanoyl-L-carnitine (C16), L-Linoleyl-L-carnitine (C18:2), L-Oleyl-L-carnitine (C18:1), L-Stearoyl-L-carnitine (C18), L-Eicosanoyl-L-carnitine (C20). As Figure 1As shown, it can be seen that in the sera of dogs suffering from pancreatitis, the acylcarnitines in the above different carbon-length groups all tend to increase compared with healthy dogs. Among them, in the C6 group, the pancreatitis dogs are 2.04 times that of the healthy dogs; in the C8:1 group, the pancreatitis dogs are 1.42 times that of the healthy dogs; in the C8 group, the pancreatitis dogs are 3.89 times that of the healthy dogs; in the C10 group, the pancreatitis dogs are 2.29 times that of the healthy dogs; in the C12:1 group, the pancreatitis dogs are 2.85 times that of the healthy dogs; in the C12 group, the pancreatitis dogs are 2.07 times that of the healthy dogs; in the C14:2 group, the pancreatitis dogs are 2.11 times that of the healthy dogs; in the C14:1 group, the pancreatitis dogs are 1.83 times that of the healthy dogs; in the C14 group, the pancreatitis dogs are 1.94 times that of the healthy dogs; in the C16:1 group, the pancreatitis dogs are 2.57 times that of the healthy dogs; in the C16 group, the pancreatitis dogs are 1.53 times that of the healthy dogs; in the C18:2 group, the pancreatitis dogs are 1.72 times that of the healthy dogs; in the C18:1 group, the pancreatitis dogs are 1.73 times that of the healthy dogs; in the C18 group, the pancreatitis dogs are 1.79 times that of the healthy dogs; in the C20 group, the pancreatitis dogs are 1.42 times that of the healthy dogs. The data from Experiment 1 show that both medium-chain acylcarnitines (6 - 12 carbon atoms) and long-chain acylcarnitines (more than 12 carbon atoms) can effectively distinguish between healthy dogs and pancreatitis dogs. Therefore, it is speculated that acylcarnitines can be used as biomarkers for pancreatitis in dogs.

[0047] Experiment 2: Urine detection in canines.

[0048] Compare the urine of healthy canines and canines with pancreatitis, and detect the content of acylcarnitines with thirteen different carbon lengths in the urine respectively. Please refer to Figure 2, a bar graph showing thirteen groups of acylcarnitines with different carbon chain lengths in the urine of healthy dogs and dogs with pancreatitis according to the present invention. The thirteen groups of acylcarnitines with different carbon chain lengths are: L-Hexanoyl carnitine (Hexanoyl L-carnitine, C6), L-Octenoyl carnitine (Octenoyl-L-carnitine, C8:1), L-Octanoyl carnitine (Octanoyl-L-carnitine, C8), L-Decenoyl carnitine (Decenoyl-L-carnitine, C10:1), L-Decanoyl carnitine (Decanoyl-L-carnitine, C10), L-Dodecenoyl carnitine (Dodecenoyl-carnitine, C12:1), L-Dodecanoyl carnitine (Dodecanoyl-L-carnitine, C12), L-Tetradecadienoyl carnitine (Tetradecadienoyl-L-carnitine, C14:2), L-Tetradecenoyl carnitine (Tetradecenoyl-L-carnitine, C14:1), L-Tetradecanoyl carnitine (Tetradecanoyl-L-carnitine, C14), L-Hexadecanoyl carnitine (Hexadecanoyl-L-carnitine, C16), L-Hexadecenoyl carnitine (Hexadecenoyl-L-carnitine, C16:1), L-Octadecadienoyl (or linolenic acid) carnitine (Linoleyl-L-carnitine, C18:2). As Figure 2As shown, it can be seen that compared with healthy dogs, the acylcarnitines in the urine of dogs suffering from pancreatitis tend to increase. Among them, the acylcarnitines in the groups of C10, C12:1, C12, and C14:2 showed a significant increase (p<0.05). The specific data for each group are as follows: In the group of C6, the pancreatitis dogs were 6.08 times that of the healthy dogs; in the group of C8:1, the pancreatitis dogs were 9.40 times that of the healthy dogs; in the group of C8, the pancreatitis dogs were 14.19 times that of the healthy dogs; in the group of C10:1, the pancreatitis dogs were 8.28 times that of the healthy dogs; in the group of C10, the pancreatitis dogs were 23.35 times that of the healthy dogs; in the group of C12:1, the pancreatitis dogs were 24.38 times that of the healthy dogs; in the group of C12, the pancreatitis dogs were 23.61 times that of the healthy dogs; in the group of C14:2, the pancreatitis dogs were 75.23 times that of the healthy dogs; in the group of C14:1, the pancreatitis dogs were 120.14 times that of the healthy dogs; in the group of C14, the pancreatitis dogs were 2086.68 times that of the healthy dogs; in the group of C16, the pancreatitis dogs were 7.30 times that of the healthy dogs; in the group of C16:1, the pancreatitis dogs were 114.01 times that of the healthy dogs; in the group of C18:2, the pancreatitis dogs were 4.08 times that of the healthy dogs. It shows that both medium-chain acylcarnitines (6-12 carbon atoms) and long-chain acylcarnitines (more than 12 carbon atoms) can effectively distinguish healthy dogs from pancreatitis dogs, and in urine, the concentration of medium / long-chain acylcarnitines in pancreatitis dogs is at least about 20 times higher than that of healthy dogs. Therefore, it can indeed be used as a biomarker for canine pancreatitis.

[0049] According to the above-mentioned Experiment 1 and Experiment 2, it can be known that there are obvious differences in the serum and urine concentrations of medium-chain acylcarnitines between healthy dogs and pancreatitis dogs. In particular, L-decanoyl carnitine (Decanoyl-L-carnitine, C10), L-dodecenoyl carnitine (Dodecenoyl-L-carnitine, C12:1), and L-dodecane (or laurel) acyl carnitine (Dodecanoyl-L-carnitine, C12) can be used as indicators for detecting urine and serum. Therefore, they are most suitable as biomarkers for diagnosing canine pancreatitis. In another embodiment, L-tetradecadienoyl carnitine (Tetradecadienoyl-L-carnitine, C14:2) in long-chain acylcarnitines is also suitable as a screening marker for urine and blood. Moreover, urine is easier to obtain from the organism than blood and does not require an invasive method to obtain blood. Therefore, as a reagent set, it can be more conveniently operated at home without going to a medical collection institution. Compared with the conventional technology of measuring cPL that requires blood collection, the measuring method or reagent set provided by the present invention is more suitable for pre-operation at the home stage.

[0050] In summary, the present invention provides a specific biomarker, namely medium-chain acylcarnitine, long-chain acylcarnitine or a combination thereof, which can correctly diagnose the probability of pancreatitis occurring in canines such as dogs.

Claims

1. A method for diagnosing canine pancreatitis, characterized in that: Include: A sample of a canine is provided; and a concentration of a biomarker in the sample is detected. If the concentration is higher than a predetermined value, the canine is determined to have a high risk of pancreatitis, wherein the predetermined value is related to the concentration of the biomarker in a healthy canine, and the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine or a combination thereof, wherein the medium-chain acylcarnitine has a chain length of 6-12 carbon atoms, and the long-chain acylcarnitine has a chain length of more than 12 carbon atoms.

2. The method according to claim 1, characterized in that The medium-chain carnitine referred to by the biomarker refers to L-decanoyl-L-carnitine (C10), L-dodecanoyl (or lauryl) carnitine (C12) or L-dodecenoyl-L-carnitine (C12:1), and the sample includes urine or blood.

3. The method according to claim 1, characterized in that The long-chain carnitine referred to by the biomarker is L-tetradecadienoyl-L-carnitine (C14:2), and the sample includes urine or blood.

4. A kit for diagnosing pancreatitis in canines, characterized in that: The invention comprises a detection reagent, which can be used to detect the concentration of a biomarker in a canine sample after being mixed with the detection reagent. The biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine or a combination thereof, wherein the medium-chain acyl group refers to a group with a chain length of 6-12 carbon atoms, and the long-chain acyl group refers to a group with a chain length of more than 12 carbon atoms.

5. The reagent set according to claim 4, characterized in that The medium-chain carnitine referred to by the biomarker refers to L-decanoyl-L-carnitine (C10), L-dodecanoyl (or lauryl) carnitine (C12), and L-dodecenoyl-L-carnitine (C12:1), and the sample includes urine or blood.

6. The reagent set according to claim 4, characterized in that The long-chain carnitine referred to by the biomarker is L-tetradecadienoyl-L-carnitine (C14:2), and the sample includes urine or blood.

7. A method of using medium-chain acylcarnitine, long-chain acylcarnitine or a combination thereof for diagnosing pancreatitis in canines, characterized in that: A medium chain acyl group refers to an acyl group having a chain length of 6 to 12 carbon atoms, and a long chain acyl group refers to an acyl group having a chain length of more than 12 carbon atoms.

8. The use according to claim 7, characterized in that It refers to the use of L-decanoyl-L-carnitine (C10), L-dodecanoyl (or lauryl) carnitine (Dodecanoyl-L-carnitine, C12) or L-dodecenoyl-L-carnitine (Dodecenoyl-L-carnitine, C12:1) for diagnosing pancreatitis in canines, and the diagnosis is performed by taking urine or blood of the canines as the specimen.

9. The use according to claim 7, characterized in that The invention refers to the use of L-tetradecadienoyl-L-carnitine (C14:2) for diagnosing pancreatitis in canines, wherein the diagnosis is performed by taking urine or blood of the canines as a specimen.

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