Method for diagnosing pancreatitis in canids, reagent kit, and use of acylcarnitine in the diagnosis of pancreatitis in canids

Abstract

Provided are a method and a reagent kit for diagnosing pancreatitis in canids, which can rapidly assist in diagnosing the possibility of pancreatitis in canids or assist in discriminating the severity of acute pancreatitis in canids by detecting the concentration of a specific biomarker in a specimen, and the use of acylcarnitine in the diagnosis of pancreatitis in canids. 【Means for solving the problem】 First, a specimen of a canid is provided. Then, the concentration of a biomarker in the specimen is detected. If the concentration is higher than a predetermined value, it is determined that the canid has a high risk of pancreatitis, and the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, where the medium-chain acyl has a chain length of 6 to 12 carbon atoms, and the long-chain acyl has a chain length exceeding 12 carbon atoms.

Description

Technical Field

[0001] The present invention relates to a method for diagnosing pancreatitis in canids and a reagent kit for diagnosing pancreatitis in canids, and particularly to a method or reagent kit for diagnosing pancreatitis in canids using a special biomarker.

Background Art

[0002] Acute pancreatitis is caused by abnormal activation of trypsinogen in the pancreas, which results in ineffective functioning of trypsin inhibition in pancreatic acinar cells, leading to autodigestion and inflammation. It is one of the most severe diseases of the digestive system in dogs. Mild pancreatitis usually presents with clinical symptoms such as mild to moderate anorexia, abdominal pain, and vomiting, and can be completely recovered with appropriate symptomatic treatment. However, severe acute pancreatitis is accompanied by obvious pancreatic necrosis and usually involves a severe systemic inflammatory response and multiple organ dysfunction, with a fatality rate that can approach 50%.

[0003] Currently, the clinical diagnosis of acute pancreatitis in dogs, especially the determination of severity, remains difficult. The currently commonly used diagnostic index is serum canine pancreatic-specific lipase (cPL), but there is still room for improvement in its specificity and sensitivity, and it is unable to clearly distinguish the severity of acute pancreatitis. In actual diagnosis, it is still necessary to include the medical history, clinical symptoms, and abdominal ultrasound results.

Summary of the Invention

Problems to be Solved by the Invention

[0004] The present invention provides a method for diagnosing or detecting pancreatitis in canids and a reagent kit for diagnosing or detecting pancreatitis in canids, which can quickly assist in diagnosing the possibility of pancreatitis in canids or assist in discriminating the severity of acute pancreatitis in canids by detecting the concentration of a special biomarker in a specimen.

Means for Solving the Problems

[0005] Based on one embodiment of the present invention, the present invention provides a method for diagnosing pancreatitis in canids. First, a specimen of a canid is provided. Then, the concentration of a biomarker in the specimen is detected. If the concentration is higher than a predetermined value, it is determined that the canid 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 refers to those having a chain length of 6 to 12 carbon atoms, and the long-chain acyl refers to those having a chain length exceeding 12 carbon atoms.

[0006] Based on one embodiment of the present invention, the present invention provides a reagent kit for diagnosing pancreatitis in canids. The kit includes a detection reagent that can be used to detect the concentration of a biomarker in a specimen of a canid after being mixed with the specimen. The biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof. The medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and the long-chain acyl refers to those having a chain length exceeding 12 carbon atoms.

[0007] Based on another embodiment of the present invention, the present invention provides the use of medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof in the diagnosis of pancreatitis in canids, where the medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and the long-chain acyl refers to those having a chain length exceeding 12 carbon atoms.

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

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

Effects of the Invention

[0010] By detecting the special biomarkers provided by the present invention, namely medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, it is possible to assist in diagnosing whether a canine animal develops pancreatitis. When it is discovered that a canine animal has a high possibility of pancreatitis, treatment can be carried out immediately, and the survival rate can be significantly improved.

Brief Description of the Drawings

[0011]

Figure 1

Figure 2

Modes for Carrying Out the Invention

[0012] To enable those skilled in the art to further understand the present invention, the following provides preferred embodiments of the present invention, and in conjunction with the drawings, the structural content of the present invention and the effects to be achieved will be described in detail.

[0013] At present, the main causes of acute pancreatitis are not clear, but the generally considered causes are high-fat diet, inappropriate diet, obesity, drugs and toxins, endocrine diseases, and genetics. Taking dogs as an example, pancreatitis often occurs in middle-aged and elderly dogs, and although in small numbers, it also occurs in dog breeds such as Miniature Schnauzers. Acylcarnitine is one of the intermediate products of fatty acid metabolism and is produced by the esterification of fatty acids and L-carnitine. Therefore, the content of acylcarnitine changes with the regulation of fatty acid metabolism. Therefore, acylcarnitine can reflect diseases related to fatty acid oxidation disorders and metabolic disorders. When the β-oxidation pathway of fatty acids is affected, the level of acylcarnitine increases significantly. As a product of the fatty acid metabolism pathway, acylcarnitine exists in body fluids such as serum and urine, and at the same time has the ability to activate information transmission related to the inflammatory response. When the content is high, it can directly affect the normal physiological functions of cells.

[0014] In order to address the lack of a good biomarker for detecting pancreatitis in canids in the prior art, the present invention provides a method for diagnosing pancreatitis in canids. First, a sample of a canid is provided. In one embodiment of the present invention, the canid is, for example, a dog, wolf, jackal or fox, etc., but is not limited thereto. In a preferred embodiment, the canid refers to a dog. On the other hand, since the sample can be obtained from a canid in various ways, the sample can be separated from the canid and made the subject of detection independently of the canid. In one embodiment, the sample may be various tissue fluids or body fluids. In a preferred embodiment, the sample refers to blood, and in particular, serum that can be obtained, for example, by blood sampling. In another preferred embodiment, the sample is urine and can be obtained by the self-urination of a canid or other invasive methods such as catheterization or bladder puncture.

[0015] Next, the concentration of the biomarker in the specimen is detected. If the concentration is higher than a predetermined value, the canine is determined to have a high risk of pancreatitis. In one embodiment of the present invention, the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof. Medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and long-chain acyl refers to those having a chain length exceeding 12 carbon atoms. In a preferred embodiment of the present invention, the biomarker refers to medium-chain acylcarnitine. When the specimen is simultaneously tested for blood and urine, the biomarker refers to decanoyl-L-carnitine (C10), dodecanoyl (or lauryl)-L-carnitine (C12), or dodecenoyl-L-carnitine (C12:1). In another optimal embodiment, the biomarker refers to long-chain acylcarnitine. When the specimen is simultaneously tested for blood and urine, the biomarker refers to tetradecadienoyl-L-carnitine (C14:2).

[0016] On the one hand, the detection of the concentration of the above-mentioned biomarker is performed using a liquid chromatography mass spectrometer (LC / MS). In this case, if the concentration is higher than a predetermined value, the canid is considered to have a high risk of pancreatitis. The aforementioned predetermined value refers to the concentration of the biomarker in the specimen of a healthy dog. Theoretically, in the case of the same canid, the predetermined value should be fixed (predetermined). This predetermined value is obtained by the following method. First, a healthy canid is prepared. The healthy canid and the canid having the disease to be detected are of the same species of organism (for example, the same pet dog). Next, a healthy specimen of the healthy canid, for example, serum or urine in the blood, is collected, and the concentration of the same biomarker (for example, dodecanoyl carnitine) in the healthy specimen is measured using an instrument. Finally, the concentration of the biomarker in the healthy specimen is compared with the concentration of the biomarker in the specimen to be tested. If the concentration of the biomarker in the specimen to be tested is greater than a certain percentage of the concentration of the biomarker in the healthy specimen (that is, greater than the predetermined value), the canid to be tested is considered to have a high risk of pancreatitis. In one embodiment of the present invention, when the specimen is serum and the concentration of the biomarker in the specimen to be tested is 2 times or more the concentration of the biomarker in the healthy specimen, the canid to be tested is considered to have a high risk of pancreatitis. In one embodiment of the present invention, when the specimen is urine and the concentration of the biomarker in the specimen to be tested is 20 times or more the concentration of the biomarker in the healthy specimen, the canid to be tested is considered to have a high risk of pancreatitis. By comparing with the predetermined value, it is possible to determine whether the canid has a high risk of pancreatitis. In addition, the high risk of pancreatitis referred to in the present invention means that the canid can be diagnosed with pancreatitis according to the conventional diagnostic method. For example, the clinical symptoms include vomiting, diarrhea, loss of appetite, abdominal pain (the symptoms satisfy at least two of them), canine-specific lipase (cPL) value: > 200 μg / L, or abdominal ultrasound examination: hypoechoic pancreatic parenchyma, pancreatic hypertrophy, blurred pancreatic margin, surrounding hyperechoic adipose tissue, and other abnormalities such as pancreatic duct / bile duct dilation and ascites, but are not limited thereto, and other diagnostic indicators sufficient to indicate that the canid is suffering from pancreatitis are also included.

[0017] In another embodiment of the present invention, the present invention provides a reagent kit for diagnosing pancreatitis in canids. The reagent kit includes a detection reagent. The detection reagent can be used to detect the concentration of a biomarker after being mixed with a sample of a canid. The biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof. Medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and long-chain acyl refers to those having a chain length exceeding 12 carbon atoms. The detection reagent includes an internal standard necessary for detecting the concentration of medium-chain / long-chain acylcarnitine. The measurement of the concentration of medium-chain / long-chain acylcarnitine can be performed by various methods known in the art, such as liquid chromatography-mass spectrometry or detection of the concentration of an internal biomarker by high-performance liquid chromatography. In one embodiment, the reagent kit of the present invention can also non-invasively diagnose medium-chain / long-chain using enzyme immunoassay (ELISA), affinity chromatography column purification method, or improved test tube binding analysis method, etc. For example, the present invention detects pancreatitis in canids using the ELISA method. First, the medium-chain / long-chain acylcarnitine of the present invention is connected to a calibration 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-chain / long-chain acylcarnitine in the sample is analyzed, for example, to determine whether there is a risk of pancreatitis in canids. The same parts as the foregoing embodiments of the related implementation methods of this embodiment, such as canids, samples, biomarkers, etc., will not be described again here.

[0018] In another embodiment of the present invention, there is provided the use of medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof in the diagnosis of pancreatitis in canids, where medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and long-chain acyl refers to those having a chain length exceeding 12 carbon atoms. The same parts as the foregoing embodiments of the related implementation methods of this embodiment, such as canids, samples, biomarkers, etc., will not be described again here.

[0019] To clearly explain the implementation method and effects of the present invention, the following will provide examples and related experiments and describe their detailed procedures.

Example

[0020] The experimental process of this example is as follows. (1) This study was reviewed and approved by the Institutional Animal Care and Use Committee of National Chiayi University (Taiwan), with approval number 111026. (2) Case collection (1) Inclusion criteria for dogs with the disease A. Clinical symptoms: including vomiting, diarrhea, loss of appetite, abdominal pain (at least two of these symptoms must be met). B. Canine pancreatic lipase (cPL) value: > 200 μg / L C. Ultrasonic examination of the entire abdomen: hypoechoic pancreatic parenchyma, pancreatic gland hypertrophy, blurred pancreatic margin, other abnormalities such as surrounding hyperechoic adipose tissue and dilation of the pancreatic duct / bile duct and ascites. (2) Exclusion criteria for dogs with the disease A. History of primary diseases of the gastrointestinal, hepatobiliary, and urinary systems B. No obvious abnormalities in pancreatic ultrasound examination and inability to clearly observe the pancreatic image by ultrasound examination. C. Clinical symptoms lasting for more than 7 days (3) Inclusion criteria for healthy dogs No abnormal clinical symptoms, and normal blood tests, abdominal ultrasound examination, and cPL value. (3) Collection of blood and urine (1) Healthy dogs: Blood is collected from the radial vein during a health check, and urine is collected by natural urination, with each collected once. (2) Sick dogs: Blood is collected from the radial vein during treatment. If natural urination is not possible, urine is collected using bladder puncture, and a sedative (Alfaxalone, 0.3 mL / kg) is used as needed. (4) Sample testing Metabolite and liposome analyses of blood and urine were performed using an Agilent 1290 UPLC coupled with 6540-QTOF (UHPLC-QTOF) mass spectrometer (Agilent Technologies, Santa Clara, CA) according to the published method [1]. To control the quality of the analysis, blank samples and pooled analytical quality control samples were inserted for analytical calibration at the start of each batch and every five samples to ensure the quality of the analysis. At the same time, at the start of each analysis, the performance of the instrument was tested using 40 synthetic standards. Each sample was analyzed three times, 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 confirmation of spectral peaks was performed by comparing them with a metabolome database [4] constructed independently. The features of potential metabolites selected by the comparison were preprocessed to remove those with more than 50% missing values, and the remaining features were obtained as half of the minimum positive value of the original data. Before performing MetaboAnalyst statistical analysis on the preprocessed data, the sum of the total peak areas was normalized, and after logarithmic transformation (log transform) and autoscaling, it was analyzed using MetaboAnalyst 5.0 [5]. For the methods cited in this paragraph, the following references can be consulted. 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. 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 / . 3.https: / / mzmine.github.io / mzmine_documentation / module_docs / lc-ms_featdet / featdet_adap_chromatogram_builder / adap-chromatogram-builder.html 4. In-house database: the National Taiwan University MetaCore Metabolomics Chemical Standard Library. 5.https: / / www.metaboanalyst.ca / docs / About.xhtml

[0021] <Experiment 1: Detection of Canidae Serum> The sera of healthy canids and canids suffering from pancreatitis were compared, and the serum contents of 15 sets of acylcarnitines with different carbon chain lengths were detected. Referring to Figure 1, Figure 1 shows a bar graph indicating the relative contents of 15 sets of acylcarnitines with different carbon chain lengths in the sera of healthy dogs and dogs suffering from pancreatitis according to the present invention. The 15 sets of acylcarnitines with the following carbon chain lengths are as follows. Hexanoyl-L-carnitine (C6), Octenoyl-L-carnitine (C8:1), Octanoyl-L-carnitine (C8), Decanoyl-L-carnitine (C10), Dodecenoyl-L-carnitine (C12:1), Dodecanoyl (or lauryl)-L-carnitine (C12), Tetradecadienoyl-L-carnitine (C14:2), Tetradecenoyl-L-carnitine (C14:1), Tetradecanoyl-L-carnitine (C14), Hexadecenoyl-L-carnitine (C16:1), Hexadecanoyl-L-carnitine (C16), Octadecadienyl (or linoleyl)-L-carnitine (C18:2), Oleyl-L-carnitine (C18:1), Octadecanoyl (or stearoyl)-L-carnitine (C18), Eicosanoyl-L-carnitine (C20). As shown in Figure 1, it can be seen that in the sera of dogs suffering from pancreatitis, all of the above-mentioned acylcarnitines with different carbon chain lengths tend to increase compared to healthy dogs.Among them, in the C6 group, the dogs suffering from pancreatitis were 2.04 times that of healthy dogs. In the C8:1 group, the dogs suffering from pancreatitis were 1.42 times that of healthy dogs. In the C8 group, the dogs suffering from pancreatitis were 3.89 times that of healthy dogs. In the C10 group, the dogs suffering from pancreatitis were 2.29 times that of healthy dogs. In the C12:1 group, the dogs suffering from pancreatitis were 2.85 times that of healthy dogs. In the C12 group, the dogs suffering from pancreatitis were 2.07 times that of healthy dogs. In the C14:2 group, the dogs suffering from pancreatitis were 2.11 times that of healthy dogs. In the C14:1 group, the dogs suffering from pancreatitis were 1.83 times that of healthy dogs. In the C14 group, the dogs suffering from pancreatitis were 1.94 times that of healthy dogs. In the C16:1 group, the dogs suffering from pancreatitis were 2.57 times that of healthy dogs. In the C16 group, the dogs suffering from pancreatitis were 1.53 times that of healthy dogs. In the C18:2 group, the dogs suffering from pancreatitis were 1.72 times that of healthy dogs. In the C18:1 group, the dogs suffering from pancreatitis were 1.73 times that of healthy dogs. In the C18 group, the dogs suffering from pancreatitis were 1.79 times that of healthy dogs. In the C20 group, the dogs suffering from pancreatitis were 1.42 times that of healthy dogs. The data of Experiment 1 indicate that both medium-chain acylcarnitines (with 6 to 12 carbon atoms) and long-chain acylcarnitines (with more than 12 carbon atoms) can effectively distinguish between healthy dogs and dogs suffering from pancreatitis. Therefore, it is presumed that acylcarnitine can be used as a biomarker for dogs suffering from pancreatitis.

[0022] <Experiment 2: Detection of Urine of Canidae Animals> The urine of healthy canids and canids suffering from pancreatitis was compared, and the contents of 13 groups of acylcarnitines with different carbon chain lengths in the urine were detected. Referring to Figure 2, Figure 2 shows a bar graph of 13 groups of acylcarnitines with different carbon chain lengths in the urine of healthy dogs and dogs suffering from pancreatitis of the present invention. The 13 groups of acylcarnitines with carbon chain lengths are as follows. Hexanoyl-L-carnitine (C6), Octenoyl-L-carnitine (C8:1), Octanoyl-L-carnitine (C8), Decenoyl-L-carnitine (C10:1), Decanoyl-L-carnitine (C10), Dodecenoyl-L-carnitine (C12:1), Dodecanoyl-L-carnitine (C12), Tetradecadienoyl-L-carnitine (C14:2), Tetradecenoyl-L-carnitine (C14:1), Tetradecanoyl-L-carnitine (C14), Hexadecanoyl-L-carnitine (C16), Hexadecenoyl-L-carnitine (C16:1), Hexadecanoyl-L-carnitine (C16), Octadecadienyl (or linoleyl)-L-carnitine (C18:2). As shown in Figure 2, it was found that compared with healthy dogs, the urine of dogs suffering from pancreatitis tended to have an increase in acylcarnitine, and the acylcarnitines in the groups of C10, C12:1, C12, and C14:2 increased significantly (p < 0.05). The specific data for each group are as follows.In the C6 group, dogs with pancreatitis were 6.08 times more likely than healthy dogs. In the C8:1 group, dogs with pancreatitis were 9.40 times more likely than healthy dogs. In the C8 group, dogs with pancreatitis were 14.19 times more likely than healthy dogs. In the C10:1 group, dogs with pancreatitis were 8.28 times more likely than healthy dogs. In the C10 group, dogs with pancreatitis were 23.35 times more likely than healthy dogs. In the C12:1 group, dogs with pancreatitis were 24.38 times more likely than healthy dogs. In the C12 group, dogs with pancreatitis were 23.61 times more likely than healthy dogs. In the C14:2 group, dogs with pancreatitis were 75.23 times more likely than healthy dogs. In the C14:1 group, dogs with pancreatitis were 120.14 times more likely than healthy dogs. In the C14 group, dogs with pancreatitis were 2086.68 times more likely than healthy dogs. In the C16 group, dogs with pancreatitis were 7.30 times more likely than healthy dogs. In the C16:1 group, dogs with pancreatitis were 114.01 times more likely than healthy dogs. In the C18:2 group, dogs with pancreatitis were 4.08 times more likely than healthy dogs. Regardless of whether it is medium-chain acylcarnitine (carbon number 6 - 12) or long-chain acylcarnitine (carbon number greater than 12), it was shown that both could effectively distinguish between healthy dogs and dogs with pancreatitis. In urine, the concentration of medium-chain / long-chain acylcarnitine in dogs with pancreatitis was at least about 20 times higher than that in healthy dogs. Therefore, it can surely be used as a biomarker for canine pancreatitis.

[0023] According to the above-mentioned Experiment 1 and Experiment 2, it is found that there are significant differences in the concentrations of medium-chain carnitine in the serum and urine of healthy dogs and dogs suffering from pancreatitis. In particular, decanoyl-L-carnitine (C10), dodecenoyl-L-carnitine (C12:1), and dodecanoyl (or lauryl)-L-carnitine (C12) can be used as indicators for detecting urine and serum, so they are optimal as biomarkers for diagnosing canine pancreatitis. In another embodiment, tetradecadienoyl-L-carnitine (C14:2) in long-chain acylcarnitine is also suitable as a screening marker for urine and blood. In addition, urine can be easily obtained from the living body compared to blood, and there is no need to obtain blood by an invasive method. Therefore, as a reagent kit, it can be more conveniently operated at home without the need to go to a medical collection institution. Compared with the conventional cPL measurement technology that requires blood sampling, the measurement method or reagent kit provided by the present invention is suitable for the home stage and can be pre-operated.

[0024] In summary, the present invention provides special biomarkers, and by using medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, the probability of pancreatitis occurrence in canids, such as dogs, can be accurately diagnosed.

Claims

1. A step of providing a sample of a Canidae animal, a step of detecting the concentration of a biomarker in the sample, and when the concentration is higher than a predetermined value, determining that the Canidae animal has a high risk of pancreatitis, wherein the predetermined value relates to the concentration of the biomarker in a healthy Canidae animal, and the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, the medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and the long-chain acyl refers to those having a chain length exceeding 12 carbon atoms. When the sample is serum, if the concentration of the biomarker is 2 times or more of the predetermined value, it is determined that the risk of pancreatitis is high. A method for diagnosing pancreatitis in a Canidae animal, when the sample is urine, if the concentration of the biomarker is 20 times or more of the predetermined value, it is determined that the risk of pancreatitis is high.

2. The medium-chain acylcarnitine referred to by the biomarker refers to decanoyl-L-carnitine, dodecanoyl-L-carnitine, or dodecenoyl-L-carnitine, and the sample includes urine or blood. The method for diagnosing pancreatitis in a Canidae animal according to Claim 1.

3. The long-chain acylcarnitine referred to by the biomarker refers to tetradecadienoyl-L-carnitine, and the sample includes urine or blood. The method for diagnosing pancreatitis in a Canidae animal according to Claim 1.

4. A reagent kit for diagnosing pancreatitis in a Canidae animal, including a detection reagent that can be used to detect the concentration of a biomarker in a sample after mixing with a sample of a Canidae animal, the biomarker refers to medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof, the medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, and the long-chain acyl refers to those having a chain length exceeding 12 carbon atoms.

5. The medium-chain acylcarnitine referred to by the biomarker refers to decanoyl-L-carnitine, dodecanoyl-L-carnitine, or dodecenoyl-L-carnitine, and the sample includes urine or blood. The reagent kit for diagnosing pancreatitis in a Canidae animal according to Claim 4.

6. The long-chain acylcarnitine referred to by the biomarker refers to tetradecadienoyl-L-carnitine, and the sample includes urine or blood. The reagent kit for diagnosing pancreatitis in a Canidae animal according to Claim 4.

7. Use of medium-chain acylcarnitine, long-chain acylcarnitine, or a combination thereof as a biomarker in the diagnosis of pancreatitis in canids, wherein the medium-chain acyl refers to those having a chain length of 6 to 12 carbon atoms, the long-chain acyl refers to those having a chain length exceeding 12 carbon atoms, and the diagnosis uses urine or blood of canids as a specimen. Claim 8 The use according to claim 7, which refers to the use of decanoyl-L-carnitine, dodecanoyl-L-carnitine, or dodecenoyl-L-carnitine in the diagnosis of pancreatitis in canids. Claim 9 The use according to claim 7, which refers to the use of tetradecadienoyl-L-carnitine in the diagnosis of pancreatitis in canids.

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