TWI833791B - Auxiliary methods for detection of primary biliary cholangitis - Google Patents

Abstract

The subject of the present invention is to provide a simple and auxiliary method for detecting primary biliary cholangitis using blood tests.

The solution of the present invention is an auxiliary method for the detection of primary biliary cholangitis, which is to detect LDL-TG, RLP-C, LDL-TG/LDL-C, sd LDL in the tested blood sample separated from the living body. -Choose at least 1 type from the group consisting of LDL-C, LDL-C/sd LDL-C, LDL-TG/sd LDL-C, total TG and total TG/total CHO. When the LDL-TG level is used as an indicator, if the LDL-TG level is higher than that of healthy people, it means that the possibility of primary biliary cholangitis is high.

Description

Auxiliary methods for detection of primary biliary cholangitis

The present invention relates to a method for assisting in the detection of primary biliary cholangitis using blood tests.

It is commonly known that primary biliary cholangitis (PBC) is diagnosed based on histological analysis, blood analysis, chronic bile retention analysis (increased ALP, γ-GTP), and anti-grainlandular antibody (AMA) positive interpretation, but the diagnosis The method is quite complex.

[Prior technical literature] [Non-patent literature]

[Non-patent document 1] Gershwin ME, et al. Molecular biology of the 2-oxo-acid dehydrogenase complexes and anti-mitochondrial antibodies. Prog Liver Dis 10:47-61, 1992.

The purpose of the present invention is to provide a simple and auxiliary method for detecting PBC using blood tests.

After in-depth research, the inventors of this case found that the tested blood samples separated from living organisms would include LDL-TG, RLP-C, LDL-TG/LDL-C, sd LDL-C, and LDL-C/sd LDL. -C, LDL-TG/sd LDL-C, total Tg and total TG/total CHO are selected as indicators to assist in the detection of PBC, thus completing the present invention.

That is, the present invention provides the following.

(1) An auxiliary method for detecting primary biliary cholangitis, which is to detect LDL-TG, RLP-C, LDL-TG/LDL-C, sd LDL-C in a blood sample separated from a living body , LDL-C/sd LDL-C, LDL-TG/sd LDL-C, total TG and total TG/total CHO.

(2) The method according to (1), wherein the LDL-TG amount is used as an index, and if the LDL-TG amount is higher than that of healthy subjects, it means that the possibility of primary biliary cholangitis is high.

(3) The method according to (1), wherein the RLP-C level is used as an index, and if the RLP-C level is higher than that of healthy subjects, it means that the possibility of primary biliary cholangitis is high.

(4) The method according to (1), wherein the LDL-TG/LDL-C ratio is used as an index, and if the LDL-TG/LDL-C ratio is higher than that in healthy subjects, it indicates the risk of primary biliary cholangitis. The probability is high.

(5) The method according to (1), wherein the sd LDL-C amount is used as an index, and if the sd LDL-C amount is higher than that of healthy subjects, it means that the possibility of primary biliary cholangitis is high.

(6) The method according to (1), wherein the LDL-C/sd LDL-C ratio is used as an index, and if the LDL-C/sd LDL-C ratio is lower than that of healthy subjects, it indicates primary biliary bile duct disease. The possibility of inflammation is high.

(7) The method according to (1), wherein the LDL-TG/sd LDL-C ratio is used as an index, and if the LDL-TG/sd LDL-C ratio is lower than that of healthy subjects, it indicates primary biliary bile duct disease. The possibility of inflammation is high.

(8) The method according to (1), wherein the total TG amount is used as an index, and if the total TG amount is higher than that of healthy subjects, it means that the possibility of primary biliary cholangitis is high.

(9) The method according to (1), wherein the total TG/total CHO ratio is used as an index. If the total TG/total CHO ratio is higher than that in healthy subjects, it indicates that the possibility of primary biliary cholangitis is high.

Figure 1 is a graph of results obtained in Comparative Example 1 described below.

Figure 2 is a graph of results obtained in Comparative Example 2 described below.

Figure 3 is a graph of results obtained in Comparative Example 3 described below.

Figure 4 is a graph of results obtained in Comparative Example 4 described below.

Figure 5 is a graph of results obtained in Comparative Example 5 described below.

Figure 6 is a graph of results obtained in Example 1 described below.

Figure 7 is a graph of results obtained in Example 2 described below.

Figure 8 is a graph of results obtained in Example 3 described below.

Figure 9 is a graph of results obtained in Example 4 described below.

Figure 10 is a graph of results obtained in Example 5 described below.

Figure 11 is a graph of results obtained in Example 6 described below.

Figure 12 is a graph of results obtained in Example 7 described below.

Figure 13 is a graph of results obtained in Example 8 described below.

Figure 14 is a graph of results obtained in Comparative Example 6 described below.

Figure 15 is a graph of results obtained in Comparative Example 7 described below.

Figure 16 is a graph of results obtained in Comparative Example 8 described below.

Figure 17 is a graph of results obtained in Example 9 described below.

Figure 18 is a graph of results obtained in Example 10 described below.

Figure 19 is a graph of results obtained in Example 11 described below.

Figure 20 is a graph of results obtained in Example 12 described below.

Figure 21 is a graph of results obtained in Example 13 described below.

Figure 22 is a graph of results obtained in Example 14 described below.

Figure 23 is a graph of results obtained in Example 15 described below.

In the present invention, "TG" refers to triglyceride (also known as "neutral fat"), "LDL-TG" refers to triglyceride in low-density lipoprotein, and "sd LDL-C" refers to small Cholesterol in particulate low-density lipoproteins, "RLP-C" refers to cholesterol in remnant-like lipoproteins, "LDL-C" refers to cholesterol in low-density lipoproteins, and "CHO" refers to cholesterol.

In addition, these abbreviations are common abbreviations in the field of blood lipids.

The blood sample system can use whole blood, serum or plasma.

Total TG, LDL-TG, sd LDL-C, RLP-C, LDL-C and total CHO in the blood sample can be detected using well-known methods, and the detection methods are also specifically described in the following examples.

In the present invention, when the total TG amount is used as an index, if the total TG amount is higher than that of healthy people, It means that the possibility of PBC is high. The cutoff value used in the determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 50~158mg/dL, preferably within the range of ±20% of 96.0mg/dL, and the best The optimal value is 96.0 mg/dL, and can be set to the value of sensitivity and specificity required by the inventor. In addition, the "cutoff value" is a value that determines the possibility of primary biliary cholangitis to be high if it is above or below this value. When processing statistically, the optimal value of the cutoff value is performed by ROC curve analysis and calculated using Youden's index.

In the present invention, when the LDL-TG amount is used as an index, if the LDL-TG amount is higher than that of healthy people, it means that the possibility of PBC is high. The cutoff value used for determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 10.0~20.5mg/dL, preferably within the range of 12.7mg/dL±20%, and the most suitable range. The optimal value is 12.7 mg/dL, and can be set to a value that matches the sensitivity and specificity required by the inventor.

In the present invention, when the sd LDL-C amount is used as an index, if the sd LDL-C amount is higher than that of healthy people, it means that the possibility of PBC is high. The cutoff value used for determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 12.4~43.9mg/dL, preferably within the range of ±20% of 21.2mg/dL. The optimal value is 21.2 mg/dL, and can be set to a value that matches the sensitivity and specificity required by the inventor.

In the present invention, when the RLP-C level is used as an index, if the RLP-C level is higher than that of healthy people, it means that the possibility of PBC is high. Determine whether the cutoff value used can be used in conjunction with this The sensitivity and specificity are appropriately set within the range required by the inventor, for example, within the range of 1.4~3.7mg/dL, preferably within the range of 2.7mg/dL±20%, and the optimal value is 2.7mg/dL, and can be set In order to cooperate with the use of the sensitivity and specificity values required by the inventor of the present invention.

In the present invention, when the LDL-TG/LDL-C ratio is used as an index, if the LDL-TG/LDL-C ratio is higher than that of healthy people, it means that the possibility of PBC is high. The cutoff value used for determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 0.104~0.193, preferably within the range of 0.134±20%, and the optimal value is 0.134, and It can be set to the value of sensitivity and specificity required by the inventor of the present invention.

In the present invention, when the total TG/total CHO ratio is used as an index, if the total TG/total CHO ratio is higher than that of healthy people, it means that the possibility of PBC is high. The cutoff value used for determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 0.229~1.171, preferably within the range of 0.523±20%, and the optimal value is 0.523, and It can be set to the value of sensitivity and specificity required by the inventor of the present invention.

In the present invention, when the LDL-C/sd LDL-C ratio is used as an index, if the LDL-C/sd LDL-C ratio is lower than that of healthy people, it means that the possibility of PBC is high. The cutoff value used for determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 2.67~7.04, preferably within the range of 5.16±20%, and the optimal value is 5.16, and It can be set to the value of sensitivity and specificity required by the inventor of the present invention.

In the present invention, when the LDL-TG/sd LDL-C ratio is used as an index, if the LDL-TG/sd LDL-C ratio is lower than that of healthy people, it means that the possibility of PBC is high. The cutoff value used for determination can be appropriately set within the range of sensitivity and specificity required by the inventor, for example, within the range of 1.12~3.04, preferably within the range of 1.74±20%, and the optimal value is 1.74, and It can be set to the value of sensitivity and specificity required by the inventor of the present invention.

Hereinafter, the present invention will be described in detail based on examples. However, the present invention is not limited only to the following examples.

In addition, the specific detection methods of each lipid used in the following examples or comparative examples are as follows.

1. Detection method of total TG

The total TG amount was measured using an automatic analysis device used at the clinical examination site, using the TG-EX "Shengken" (enzyme method) for triglyceride detection reagent (manufactured by Denka Sanken Co., Ltd.).

2.Detection method of LDL-TG

The LDL-TG level was measured using an automatic analysis device used at clinical examination sites, using the LDL-triglyceride detection reagent LDLTG-EX "SEIKEN" (manufactured by Denka Biotech Co., Ltd.).

3.Detection method of sd LDL-C

The sd LDL-C level was measured using an automatic analyzer using sd LDL-Cholesterol detection reagent sd LDL-EX "Seiken" (manufactured by Denka Seikan Co., Ltd.).

4.Detection method of RLP-C

The amount of RLP-C is measured using an automatic analysis device and a reagent for RLP-cholesterol detection. The reagent for RLP-cholesterol detection was prepared according to 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.Detection method of LDL-C

The LDL-C level is measured using an automatic analysis device used at clinical examination sites, using the automatic analysis reagent for LDL-cholesterol detection "Seikan" LDL-EX(N) (direct method) (manufactured by Denka Seikan Co., Ltd.).

6. Detection method of total CHO

The total CHO amount was measured using an automatic analysis device used at clinical examination sites, using the automatic analysis reagent "Shengken" T-CHO(S) (enzyme method) (manufactured by Denka Sanken Co., Ltd.) as a cholesterol detection reagent.

Furthermore, the comparison between the two groups is based on the Shapiro-Wilk test, and in the case of non-parametric (non-parametric), the Wilcoxon-Mann-Whitney test is performed, and in the case of parametric (parametric), the isodispersion test is performed, and the Student's t-test or Welch's t-test.

Comparative example 1 (LDL-C)

Blood was drawn from a group of 96 people, consisting of 48 uncorresponding healthy subjects (labeled "Health" in the figure) and 48 PBC patients, and the LDL-C amount in the collected blood was measured, and comparisons were made between each group. The comparison result chart is shown in Figure 1. As a result, there was no difference in LDL-C levels between the PBC patient group and the healthy group.

Comparative example 2 (HDL3-C)

Blood was drawn from a group of 96 people, consisting of 48 unmatched healthy subjects (labeled "Health" in the figure) and 48 PBC patients. The amount of HDL3-C in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 2. As a result, there was no difference in HDL3-C levels between the PBC patient group and the healthy group.

Comparative Example 3 (Total CHO)

Blood was drawn from a group of 96 people consisting of 48 unmatched healthy subjects (marked "Health" in the figure) and 48 PBC patients. The amount of T-CHO in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 3. As a result, although the amount of T-CHO in the PBC patient group tended to be higher than that in the healthy group, there was no statistically significant difference.

Comparative example 4 (HDL-C)

Blood was drawn from a group of 96 people, consisting of 48 unmatched healthy subjects (labeled "Health" in the figure) and 48 PBC patients, and the amount of HDL-C in the collected blood was measured, and comparisons were made between each group. The comparison result chart is shown in Figure 4. As a result, PBC patients Although the HDL-C amount of the group showed a tendency to be higher than that of the healthy group, there was no statistically significant difference.

Comparative example 5 (HDL2-C)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy subjects (labeled "Health" in the figure) and 48 PBC patients. The amount of HDL2-C in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 5. As a result, although the HDL2-C level in the PBC patient group tended to be higher than that in the healthy group, there was no statistically significant difference.

Example 1 (Total TG)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy subjects (labeled "Health" in the figure) and 48 PBC patients. The total TG amount in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 6. As a result, the total TG amount in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0001).

Example 2 (LDL-TG)

Blood was drawn from a group of 96 people, consisting of 48 unmatched healthy subjects (labeled "Health" in the figure) and 48 PBC patients, and the amount of LDL-TG in the collected blood was measured, and comparisons were made between each group. The comparison result chart is shown in Figure 7. As a result, the amount of LDL-TG in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0001).

Example 3 (sd LDL-C)

48 PBC patients from uncorresponding healthy subjects (labeled "Health" in the figure) A group of 96 people composed of 48 people drew blood, and the amount of sd LDL-C in the collected blood was measured, and comparisons were made between groups. The comparison result chart is shown in Figure 8. As a result, the amount of sd LDL-C in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.001).

Example 4 (RLP-C)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy subjects (labeled "Health" in the figure) and 48 PBC patients. The amount of RLP-C in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 9. As a result, the amount of RLP-C in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0001).

Example 5 (LDL-TG/LDL-C)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy individuals (marked "Health" in the figure) and 48 PBC patients, and the LDL-TG and LDL-C levels in the collected blood were measured and calculated. LDL-TG/LDL-C ratio was used to compare each group. The comparison result chart is shown in Figure 10. As a result, the LDL-TG/LDL-C ratio in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0001).

Example 6 (total TG/total CHO)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy subjects (marked "Health" in the figure) and 48 PBC patients. The total TG amount and total CHO amount in the collected blood were measured, and the total TG was calculated. /total CHO ratio to compare each group. The comparison result chart is shown in Figure 11. As a result, the total TG/total CHO ratio in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0005).

Example 7 (LDL-C/sd LDL-C)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy individuals (marked as "Health" in the figure) and 48 PBC patients, and the LDL-C amount and sd LDL-C amount in the collected blood were measured. Calculate the LDL-C/sd LDL-C ratio and compare each group. The comparison result chart is shown in Figure 12. As a result, the LDL-C/sd LDL-C ratio of the PBC patient group was statistically significantly lower than that of the healthy group (p<0.0001).

Example 8 (LDL-TG/sd LDL-C)

Blood was drawn from a group of 96 people consisting of 48 uncorresponding healthy subjects (marked "Health" in the figure) and 48 PBC patients, and the LDL-TG and sd LDL-C levels in the collected blood were measured. Calculate the LDL-TG/sd LDL-C ratio and compare each group. The comparison result chart is shown in Figure 13. As a result, the LDL-TG/sd LDL-C ratio in the PBC patient group was statistically significantly lower than that in the healthy group (p<0.005).

Comparative Example 6 (Total CHO)

Blood was drawn from a group of 52 people, consisting of 27 uncorresponding healthy subjects (labeled "Health" in the figure) and 25 PBC patients. The total CHO amount in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 14. As a result, the total CHO amount in the PBC patient group was no different from that in the healthy group.

Comparative example 7 (HDL-C)

Blood was drawn from a group of 52 people, consisting of 27 uncorresponding healthy subjects (labeled "Health" in the figure) and 25 PBC patients, and the HDL-C amount in the collected blood was measured, and comparisons were made between each group. The comparison result chart is shown in Figure 15. As a result, patients with PBC There was no difference in HDL-C levels between the patient group and the healthy group.

Comparative example 8 (HDL2-C)

Blood was drawn from a group of 52 people, consisting of 27 unmatched healthy subjects (labeled "Health" in the figure) and 25 PBC patients. The amount of HDL2-C in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 16. As a result, there was no difference in HDL2-C levels between the PBC patient group and the healthy group.

Example 9 (Total TG)

Blood was drawn from a group of 52 people, consisting of 27 unmatched healthy subjects (labeled "Health" in the figure) and 25 PBC patients. The total TG amount in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 17. As a result, the total TG amount in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.005).

Example 10 (LDL-TG)

Blood was drawn from a group of 52 people, consisting of 27 uncorresponding healthy subjects (labeled "Health" in the figure) and 25 PBC patients, and the LDL-TG amount in the collected blood was measured, and comparisons were made between each group. The comparison result chart is shown in Figure 18. As a result, the amount of LDL-TG in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0001).

Example 11 (RLP-C)

Blood was drawn from a group of 52 people, consisting of 27 unmatched healthy subjects (labeled "Health" in the figure) and 25 PBC patients. The amount of RLP-C in the collected blood was measured and compared between each group. The comparison result chart is shown in Figure 19. As a result, patients with PBC The amount of RLP-C in the patient group was statistically significantly higher than that in the healthy group (p<0.0005).

Example 12 (LDL-TG/LDL-C)

Blood was drawn from a group of 52 people consisting of 27 uncorresponding healthy subjects (marked "Health" in the figure) and 25 PBC patients, and the LDL-TG and LDL-C levels in the collected blood were measured and calculated. LDL-TG/LDL-C ratio was used to compare each group. The comparison result chart is shown in Figure 20. As a result, the amount of LDL-TG/LDL-C in the PBC patient group was statistically significantly higher than that in the healthy group (p<0.0001).

Example 13 (Total TG/T-CHO)

Blood was drawn from a group of 52 people consisting of 27 uncorresponding healthy subjects (marked "Health" in the figure) and 25 PBC patients. The total TG amount and total CHO amount in the collected blood were measured, and the total TG was calculated. /total CHO ratio to compare each group. The comparison result chart is shown in Figure 21. As a result, the amount of total TG/total CHO in the PBC patient group was significantly higher than that in the healthy group (p<0.005).

Example 14 (LDL-C/sd LDL-C)

Blood was drawn from a group of 52 people, consisting of 27 uncorresponding healthy subjects (marked "Health" in the figure) and 25 PBC patients, and the LDL-C amount and sd LDL-C amount in the collected blood were measured. Calculate the LDL-C/sd LDL-C ratio and compare each group. The comparison result chart is shown in Figure 22. As a result, the LDL-C/sd LDL-C ratio of the PBC patient group was statistically significantly lower than that of the healthy group (p<0.005).

Example 15 (LDL-TG/sd LDL-C)

Blood was drawn from a group of 52 people, including 27 uncorresponding healthy subjects (marked "Health" in the figure) and 25 PBC patients, and the LDL-TG and sd LDL-C levels in the collected blood were measured. Calculate the LDL-TG/sd LDL-C ratio and compare each group. The comparison result chart is shown in Figure 23. As a result, the LDL-TG/sd LDL-C ratio in the PBC patient group was statistically significantly lower than that in the healthy group (p<0.01).

Claims (1)

An auxiliary method for detecting primary biliary cholangitis, which is to detect the LDL-TG/LDL-C ratio as an indicator in a blood sample separated from a living body. If the LDL-TG/LDL-C ratio is higher than that in healthy subjects, It indicates a high possibility of primary biliary cholangitis.

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