JP2023063389A - Method for assisting in detection of primary bile cholangitis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for assisting in detecting primary bile cholangitis simply by blood test.

SOLUTION: A method for assisting in the detection of primary bile cholangitis comprises measuring at least one kind selected from a group consisting of LDL-TG, RLP-C, LDL-TG/LDL-C, sd LDL-C, LDL-C/sd LDL-C, LDL-TG/sd LDL-C, total TG and total TG/total CHO in a test blood sample separated from a living body. When an amount of LDL-TG is used as an index, a higher amount of LDL-TG than that of a healthy person indicates a higher possibility of the primary bile cholangitis.

SELECTED DRAWING: Figure 7

COPYRIGHT: (C)2023,JPO&INPIT

Description

The present invention relates to a blood test to aid in the detection of primary biliary cholangitis.

Traditionally, primary biliary cholangitis (PBC) was diagnosed based on histological findings, chronic cholestasis findings (ALP, γ-GTP elevation), and antimitochondrial antibody (AMA) positive findings on blood findings. However, the diagnostic method was complicated.

Gershwin ME, et al. Molecular biology of the 2-oxo-acid dehydrogenase complexes and anti-mitochondrial antibodies. Prog Liver Dis 10:47-61, 1992.

An object of the present invention is to provide a method for easily assisting detection of PBC by a blood test.

As a result of intensive research, the inventors of the present application found that LDL-TG, RLP-C, LDL-TG/LDL-C, sd LDL-C, LDL-C/sd LDL in a test blood sample separated from a living body -C, LDL-TG/sd LDL-C, total TG and total TG/total CHO. completed the invention.

That is, the present invention provides the following.
(1) LDL-TG, RLP-C, LDL-TG/LDL-C, sd LDL-C, LDL-C/sd LDL-C, LDL-TG/sd in a test blood sample separated from a living body A method for assisting detection of primary biliary cholangitis, comprising measuring at least one selected from the group consisting of LDL-C, total TG and total TG/total CHO.
(2) The method according to (1), wherein the LDL-TG level is used as an index, and a higher LDL-TG level than that in a healthy subject indicates a high possibility of primary biliary cholangitis.
(3) The method according to (1), wherein the RLP-C level is used as an index, and a higher RLP-C level than in healthy subjects indicates a high possibility of primary biliary cholangitis.
(4) Using the LDL-TG/LDL-C ratio as an index, a higher LDL-TG/LDL-C ratio than in healthy subjects indicates a high possibility of primary biliary cholangitis (1 ) described method.
(5) The method according to (1), wherein the sd LDL-C level is used as an indicator, and a higher sd LDL-C level than in healthy subjects indicates a high possibility of primary biliary cholangitis.
(6) Using the LDL-C/sd LDL-C ratio as an index, a lower LDL-C/sd LDL-C ratio than in healthy subjects indicates a high possibility of primary biliary cholangitis. (1) The method described.
(7) Using the LDL-TG/sd LDL-C ratio as an index, a lower LDL-TG/sd LDL-C ratio than in healthy subjects indicates a high possibility of primary biliary cholangitis. (1) The method described.
(8) The method according to (1), wherein the total TG level is used as an indicator, and a higher total TG level than in healthy subjects indicates a high possibility of primary biliary cholangitis.
(9) The method according to (1), wherein the ratio of total TG/total CHO is used as an index, and a higher ratio of total TG/total CHO than in healthy subjects indicates a high possibility of primary biliary cholangitis. .

It is a figure which shows the result obtained by the following comparative example 1. FIG. It is a figure which shows the result obtained by the following comparative example 2. FIG. It is a figure which shows the result obtained by the following comparative example 3. FIG. It is a figure which shows the result obtained by the following comparative example 4. FIG. It is a figure which shows the result obtained by the following comparative example 5. FIG. It is a figure which shows the result obtained in the following Example 1. FIG. It is a figure which shows the result obtained in the following Example 2. FIG. It is a figure which shows the result obtained in the following Example 3. FIG. It is a figure which shows the result obtained by following Example 4. FIG. It is a figure which shows the result obtained by following Example 5. FIG. It is a figure which shows the result obtained by following Example 6. FIG. It is a figure which shows the result obtained by following Example 7. FIG. It is a figure which shows the result obtained by following Example 8. FIG. It is a figure which shows the result obtained by the following comparative example 6. FIG. It is a figure which shows the result obtained by the following comparative example 7. FIG. It is a figure which shows the result obtained by the following comparative example 8. FIG. It is a figure which shows the result obtained in following Example 9. FIG. It is a figure which shows the result obtained by following Example 10. FIG. It is a figure which shows the result obtained by following Example 11. FIG. It is a figure which shows the result obtained by following Example 12. FIG. It is a figure which shows the result obtained by following Example 13. FIG. It is a figure which shows the result obtained by following Example 14. FIG. It is a figure which shows the result obtained by following Example 15. FIG.

In the present invention, "TG" is triglyceride (also referred to as neutral fat), "LDL-TG" is triglyceride in low density lipoprotein, "sd LDL-C" is cholesterol in small particle low density lipoprotein, "RLP -C" means cholesterol in remnant-like lipoproteins, "LDL-C" means cholesterol in low-density lipoproteins, and "CHO" means cholesterol. These abbreviations are commonly used in the field of blood lipids.

Whole blood, serum or plasma can be used as the blood sample.

Total TG, LDL-TG, sd LDL-C, RLP-C, LDL-C and total CHO in blood samples can be measured by well-known methods, and the measurement methods are also specifically described in the examples below. It is

In the present invention, when the total TG amount is used as an index, a higher total TG amount than in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention. %, the optimum value is 96.0 mg/dL, and the value can be set according to the sensitivity and specificity desired by the person using the present invention. The "cutoff value" is a value that determines that there is a high possibility of primary biliary cholangitis if it is above or below this value. As statistical processing, the optimum cutoff value was calculated by Youden's index after analyzing the ROC curve.

In the present invention, when the LDL-TG level is used as an index, a higher LDL-TG level than that in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention, for example, within the range of 10.0 to 20.5 mg / dL, preferably 12.7 mg / dL ± It is within the range of 20%, the optimum value is 12.7 mg/dL, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

In the present invention, when the sd LDL-C level is used as an index, a higher sd LDL-C level than in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention, for example, within the range of 12.4 to 43.9 mg / dL, preferably 21.2 mg / dL The range is ±20%, the optimum value is 21.2 mg/dL, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

In the present invention, when the RLP-C amount is used as an index, a higher RLP-C amount than that in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention, for example, within the range of 1.4 to 3.7 mg / dL, preferably 2.7 mg / dL ± It is within the range of 20%, the optimum value is 2.7 mg/dL, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

In the present invention, when the LDL-TG/LDL-C ratio is used as an index, a higher LDL-TG/LDL-C ratio than healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within the range according to the sensitivity and specificity required by the person using the present invention, for example, within the range of 0.104 to 0.193, preferably within the range of 0.134 ± 20% The optimum value is 0.134, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

In the present invention, when the total TG/total CHO ratio is used as an index, a higher total TG/total CHO ratio than that in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention. The optimum value is 0.523, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

In the present invention, when the LDL-C/sd LDL-C ratio is used as an index, a lower LDL-C/sdLDL-C ratio than that in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention, for example, within the range of 2.67 to 7.04, preferably within the range of 5.16 ± 20% The optimum value is 5.16, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

In the present invention, when the LDL-TG/sd LDL-C ratio is used as an index, a lower LDL-TG/sd LDL-C ratio than in healthy subjects indicates a high possibility of PBC. The cut-off value used for determination can be appropriately set within a range according to the sensitivity and specificity required by the person using the present invention, for example, within the range of 1.12 to 3.04, preferably within the range of 1.74 ± 20% The optimum value is 1.74, and the value can be set according to the sensitivity and specificity desired by the person using the present invention.

EXAMPLES The present invention will be specifically described below based on examples. However, the present invention is not limited to the following examples.

Specific methods for measuring each lipid employed in the following Examples and Comparative Examples were as follows.
1. Measurement method of total TG The amount of total TG was measured with TG-EX “SEIKEN” (enzyme method) (manufactured by Denka Seiken Co., Ltd.), which is a reagent for measuring triglycerides, using an automatic analyzer used in clinical examinations.

2. How to measure LDL-TG
The amount of LDL-TG was measured with LDLTG-EX “SEIKEN” (manufactured by Denka Seiken Co., Ltd.), which is a reagent for measuring LDL-triglyceride, using an automatic analyzer used in clinical examinations.

3. How to measure sd LDL-C
The amount of sd LDL-C was measured using an automatic analyzer using sd LDL-EX "SEIKEN" (manufactured by Denka Seiken Co., Ltd.), which is a reagent for measuring sd LDL-cholesterol.

4. How to measure RLP-C
The amount of RLP-C was measured with an RLP-cholesterol measuring reagent using an automatic analyzer. A reagent for measuring RLP-cholesterol was prepared by the method described in the following references.
(Reference) Hirao Y, Nakajima K, Machida T, Murakami M, Ito Y. Development of a Novel Homogeneous Assay for Remnant Lipoprotein Particle Cholesterol. The Journal of Applied Laboratory Medicine 2018;03:26-36

5. How to measure LDL-C
The amount of LDL-C was measured using an automatic analyzer used in clinical testing, which is a reagent for measuring LDL-cholesterol. ).

6. Measurement method of total CHO The total CHO amount was measured using an automatic analyzer used in clinical testing, using an automatic analysis reagent "Seiken" T-CHO (S) (enzyme method) (Denka Seiken), which is a reagent for measuring cholesterol. company).

For comparison between the two groups, the Wilcoxon-Mann-Whitney test was performed when the Shapiro-Wilk test was nonparametric, and when the test was parametric, the Student's t-test or Welch's test was performed after performing the equal variance test. A t-test was performed.

Comparative Example 1 (LDL-C)
The amount of LDL-C in the blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, there was no difference in the amount of LDL-C between the PBC patient group and the healthy subject group.

Comparative Example 2 (HDL3-C)
The HDL3-C levels in the blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients were measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, there was no difference in the amount of HDL3-C between the PBC patient group and the healthy subject group.

Comparative Example 3 (total CHO)
The amount of T-CHO in the blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (indicated as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the PBC patient group tended to have a higher amount of T-CHO than the healthy subject group, but no statistically significant difference was observed.

Comparative Example 4 (HDL-C)
The amount of HDL-C in the blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the PBC patient group tended to have higher HDL-C levels than the healthy subject group, but no statistically significant difference was observed.

Comparative Example 5 (HDL2-C)
The amount of HDL2-C in the blood collected from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the PBC patient group tended to have higher HDL2-C levels than the healthy subject group, but no statistically significant difference was observed.

Example 1 (Total TG)
The total amount of TG in the blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (indicated as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the total TG amount was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0001).

Example 2 (LDL-TG)
LDL-TG levels in blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients were measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the LDL-TG level was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0001).

Example 3 (sdLDL-C)
The amount of sd LDL-C in the blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the sd LDL-C amount was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.001).

Example 4 (RLP-C)
The amount of RLP-C in blood sampled from a total of 96 unmatched groups consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the amount of RLP-C was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0001).

Example 5 (LDL-TG/LDL-C)
LDL-TG and LDL-C levels in the blood collected from a total of 96 unmatched subjects, consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients, were measured and LDL-TG/LDL-TG/ The LDL-C ratio was calculated and compared between each group. A comparative graph is shown in FIG. As a result, the LDL-TG/LDL-C ratio was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0001).

Example 6 (total TG/total CHO)
The total TG/total CHO ratio was obtained by measuring the total TG and total CHO levels in the blood collected from a total of 96 unmatched subjects, consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients. was calculated and compared for each group. A comparative graph is shown in FIG. As a result, the total TG/total CHO ratio was statistically significantly higher in the PBC patient group than in the healthy control group (p<0.0005).

Example 7 (LDL-C/sd LDL-C)
LDL-C and sd LDL-C levels in the blood collected from a total of 96 unmatched subjects, consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients, were measured and LDL-C The /sd LDL-C ratio was calculated and compared between each group. A comparative graph is shown in FIG. As a result, the LDL-C/sd LDL-C ratio was statistically significantly lower in the PBC patient group than in the healthy subject group (p<0.0001).

Example 8 (LDL-TG/sd LDL-C)
LDL-TG levels and sd LDL-C levels in the blood collected from a total of 96 unmatched subjects, consisting of 48 healthy subjects (denoted as Health in the figure) and 48 PBC patients, were measured and LDL-TG levels were measured. The /sd LDL-C ratio was calculated and compared between each group. A comparative graph is shown in FIG. As a result, the LDL-TG/sd LDL-C ratio was statistically significantly lower in the PBC patient group than in the healthy subject group (p<0.005).

Comparative Example 6 (total CHO)
The total amount of CHO in the blood sampled from a total of 52 unmatched groups consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, there was no difference in the total amount of CHO between the PBC patient group and the healthy control group.

Comparative Example 7 (HDL-C)
The amount of HDL-C in the blood sampled from a total of 52 unmatched groups consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, there was no difference in the amount of HDL-C between the PBC patient group and the healthy subject group.

Comparative Example 8 (HDL2-C)
The amount of HDL2-C in the blood sampled from a total of 52 unmatched groups consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, there was no difference in the amount of HDL2-C between the PBC patient group and the healthy subject group.

Example 9 (Total TG)
The total amount of TG in the blood sampled from a total of 52 unmatched groups consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the total TG amount was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.005).

Example 10 (LDL-TG)
The amount of LDL-TG in the blood sampled from a total of 52 unmatched groups consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the LDL-TG level was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0001).

Example 11 (RLP-C)
The amount of RLP-C in the blood sampled from a total of 52 unmatched groups consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients was measured, and the respective groups were compared. A comparative graph is shown in FIG. As a result, the amount of RLP-C was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0005).

Example 12 (LDL-TG/LDL-C)
LDL-TG and LDL-C levels in the blood collected from a total of 52 unmatched subjects, consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients, were measured and LDL-TG/LDL-TG levels were measured. The LDL-C ratio was calculated and compared between each group. A comparative graph is shown in FIG. As a result, the LDL-TG/LDL-C amount was statistically significantly higher in the PBC patient group than in the healthy subject group (p<0.0001).

Example 13 (Total TG/T-CHO)
The total TG/total CHO ratio was determined by measuring the total TG and total CHO levels in the blood collected from a total of 52 unmatched subjects, consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients. was calculated and compared for each group. A comparative graph is shown in FIG. As a result, the total TG/total CHO amount was significantly higher in the PBC patient group than in the healthy control group (p<0.005).

Example 14 (LDL-C/sd LDL-C)
LDL-C and sd LDL-C levels in the blood collected from a total of 52 unpaired subjects, consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients, were measured. The /sd LDL-C ratio was calculated and compared between each group. A comparative graph is shown in FIG. As a result, the LDL-C/sd LDL-C ratio was statistically significantly lower in the PBC patient group than in the healthy subject group (p<0.005).

Example 15 (LDL-TG/sd LDL-C)
LDL-TG and sd LDL-C levels in the blood collected from a total of 52 unmatched subjects, consisting of 27 healthy subjects (denoted as Health in the figure) and 25 PBC patients, were measured and LDL-TG levels were measured. The /sd LDL-C ratio was calculated and compared between each group. A comparative graph is shown in FIG. As a result, the LDL-TG/sd LDL-C ratio was statistically significantly lower in the PBC patient group than in the healthy subject group (p<0.01).

As a result of intensive research, the inventors of the present application have found that it is possible to assist the detection of PBC using LDL-TG/LDL-C as an index in a test blood sample separated from a living body. We have completed the present invention.

That is, the present invention includes measuring LDL-TG/LDL-C in a test blood sample separated from a living body, and a higher LDL-TG/LDL-C ratio than that of a healthy subject indicates primary disease. Kind Code: A1 Methods are provided to assist in detecting primary biliary cholangitis that is indicative of probable biliary cholangitis.

Claims (9)

LDL-TG, RLP-C, LDL-TG/LDL-C, sdLDL-C, LDL-C/sdLDL-C, LDL-TG/sdLDL-C, total TG in test blood samples isolated from living organisms and total TG/total CHO, a method for assisting detection of primary biliary cholangitis. 2. The method according to claim 1, wherein the LDL-TG level is used as an indicator, and a higher LDL-TG level than that in a healthy subject indicates a high possibility of primary biliary cholangitis. 2. The method according to claim 1, wherein the RLP-C level is used as an index, and a higher RLP-C level than that in a healthy subject indicates a high possibility of primary biliary cholangitis. The LDL-TG/LDL-C ratio is used as an index, and a higher LDL-TG/LDL-C ratio than healthy subjects indicates a high possibility of primary biliary cholangitis. Method. 2. The method according to claim 1, wherein the sdLDL-C level is used as an index, and a higher sdLDL-C level than in healthy subjects indicates a high possibility of primary biliary cholangitis. The LDL-C/sdLDL-C ratio is used as an index, and a lower LDL-C/sdLDL-C ratio than that in a healthy subject indicates a high possibility of primary biliary cholangitis. Method. The LDL-TG/sdLDL-C ratio is used as an index, and a lower LDL-TG/sdLDL-C ratio than that in healthy subjects indicates a high possibility of primary biliary cholangitis. Method. 2. The method according to claim 1, wherein the total TG amount is used as an index, and a higher total TG amount than in healthy subjects indicates a high possibility of primary biliary cholangitis. 2. The method according to claim 1, wherein the total TG/total CHO ratio is used as an index, and a higher total TG/total CHO ratio than that in healthy subjects indicates a high possibility of primary biliary cholangitis.

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