CN105716917B

CN105716917B - collecting liquid for protecting and stabilizing hemoglobin in feces and application thereof

Info

Publication number: CN105716917B
Application number: CN201610045537.9A
Authority: CN
Inventors: V·马迪; 高飞; 陆维克; 陈金树
Original assignee: Hangzhou Aotai Biotechnology Ltd By Share Ltd
Current assignee: Hangzhou Aotai Biotechnology Ltd By Share Ltd
Priority date: 2016-01-22
Filing date: 2016-01-22
Publication date: 2020-01-31
Anticipated expiration: 2036-01-22
Also published as: CN105716917A

Abstract

The invention discloses collecting solutions for protecting and stabilizing hemoglobin in feces and application thereof, wherein the collecting solutions comprise 25-45 g/L chelating agent, 0.2-1 g/L preservative, 1-10 g/L sealant, 6-12 g/L buffer agent and 25-30 g/L polyethylene glycol, wherein the chelating agent is a mixture of two of HIDS, EDTA and sodium tripolyphosphate, and the collecting solutions can be uniformly mixed according to the proportion of 50mg:2ml and then can be placed at normal temperature (7 days), 2-8 ℃ (1 month) and-20 ℃ (6 months) for storage.

Description

collecting liquid for protecting and stabilizing hemoglobin in feces and application thereof

Technical Field

The invention belongs to the technical field of biological application, and particularly relates to collecting solutions for protecting and stabilizing hemoglobin in feces and application thereof.

Background

The clinical research proves that 87% of patients with digestive tract tumors have positive occult blood tests, and the trace hemoglobin in the excrement is an abnormal phenomenon which can be detected only by in the early colorectal cancer patients, so that the detection of the trace hemoglobin in the excrement has important significance for detecting the trace hemoglobin in the excrement, early cancer patients are discovered and treated early, the long-term survival rate of the early cancer patients can reach 91%, and even anus with physiological functions of the focus limited patients can be reserved, thereby greatly reducing the pain of the patients and improving the life quality 6335, and the advanced countries such as the United states and the like recommend that patients with digestive tract occult blood is larger than 50 ml daily, have many diseases such as colorectal cancer, colitis, peptic ulcer, intestinal polyp, gastrointestinal diverticulum and the like, have no symptoms in early stages, and only have fecal occult blood.

The biggest bottleneck for detecting fecal occult blood in the market at present is that the storage time of the fecal collecting fluid is not long, and particularly, the antigen titer is reduced quickly for a sample with lower hemoglobin, but for a detection mechanism or personnel, the collected fresh sample is usually required to be placed for 1 day and days or more for detection, and the requirement on the fecal collecting fluid is very high.

Disclosure of Invention

The object of the present invention is to solve the above problems and to provide solution formulations that can protect and stabilize hemoglobin in feces.

The collecting liquid for protecting and stabilizing hemoglobin in feces comprises a chelating agent, a preservative, a sealing agent, a buffering agent and polyethylene glycol, wherein the concentration of each component in the solution is respectively 25-45 g/L, 0.2-1 g/L, 1-10 g/L, 6-12 g/L and 25-30 g/L.

The pH value of the collecting liquid for protecting and stabilizing hemoglobin in feces is 9.4-9.5.

The chelating agent is a mixture of two of 3-hydroxy-2, 2' imino disuccinic acid tetrasodium (HIDS), Ethylene Diamine Tetraacetic Acid (EDTA) and sodium tripolyphosphate.

The preservative is a mixture of Proclin 150 and Proclin 300;

the sealant is kinds of calf serum albumin and casein;

the buffer is of Tris (hydroxymethyl) aminomethane, disodium hydrogen phosphate and borax;

preferably, the collecting solution for protecting and stabilizing hemoglobin in feces consists of tetrasodium 3-hydroxy-2, 2' -iminodisuccinate (HIDS), ethylenediaminetetraacetic acid (EDTA), Proclin300, Proclin 150, Tris (hydroxymethyl) aminomethane, calf serum albumin, polyethylene glycol and purified water, wherein the concentrations of HIDS, EDTA, Proclin300, Proclin 150, Tris, calf serum albumin and polyethylene glycol in the solution are respectively 35g/L, 10g/L, 0.2g/L, 6.05g/L, 1g/L and 25 g/L.

Another objects of the present invention are to provide the use of the above-described collection for pretreatment prior to fecal testing.

The invention can store fresh excrement and the collected liquid at normal temperature (7 days), 2-8 ℃ (1 month) and-20 ℃ (6 months) after being evenly mixed according to the proportion of 50mg to 2 ml.

The method for preparing the fecal occult blood collecting fluid comprises the following specific steps:

1) weighing the components according to the formula;

2) firstly, 700ml of purified water is added into a container, and the components in the step 1) are added in sequence according to the following sequence: after uniformly mixing a buffering agent, a sealing agent, a chelating agent, polyethylene glycol and a preservative, fixing the volume to 1L, and adjusting the pH to 9.4-9.5 to obtain the fecal occult blood collecting fluid.

The collecting liquid can make the hemoglobin in the excrement more stable in the protective liquid and the storage time longer.

Detailed Description

The following examples are intended to illustrate the invention without limiting it.

Comparative example 1:

adding 70mL of purified water into a container, then sequentially adding 0.605g of Tris (hydroxymethyl) aminomethane, 1g of calf serum albumin, 3.5g of 3-hydroxy-2, 2' tetrasodium iminodisuccinate (HIDS), 2.5g of polyethylene glycol, 0.02mL of Proclin 150 and 0.02mL of Proclin300, uniformly mixing, then fixing the volume to 100mL, and adjusting the pH to 9.4 to obtain the fecal occult blood collecting fluid.

Comparative example 2:

adding 70mL of purified water into a container, then sequentially adding 0.605g of Tris (hydroxymethyl) aminomethane, 1g of calf serum albumin, 1g of Ethylene Diamine Tetraacetic Acid (EDTA), 2.5g of polyethylene glycol, 0.02mL of Proclin 150 and 0.02mL of Proclin300, uniformly mixing, fixing the volume to 100mL, and adjusting the pH value to 9.4 to obtain the fecal occult blood collecting fluid.

Example 1:

adding 70mL of purified water into a container, then sequentially adding 0.605g of Tris (hydroxymethyl) aminomethane, 1g of calf serum albumin, 3.5g of 3-hydroxy-2, 2' tetrasodium iminodisuccinate (HIDS), 1g of Ethylene Diamine Tetraacetic Acid (EDTA), 2.5g of polyethylene glycol, 0.02mL of Proclin 150 and 0.02mL of Proclin300, uniformly mixing, fixing the volume to 100mL, and adjusting the pH to 9.4 to obtain the fecal occult blood collecting solution.

Example 2:

adding 70mL of purified water into a container, then sequentially adding 0.6g of Tris (hydroxymethyl) aminomethane, 0.5g of calf serum albumin, 1g of tetrasodium 3-hydroxy-2, 2' -iminodisuccinate (HIDS), 1.5g of Ethylene Diamine Tetraacetic Acid (EDTA), 2.5g of polyethylene glycol, 0.01g of Proclin 150 and 0.01g of Proclin300, uniformly mixing, fixing the volume to 100mL, and adjusting the pH to 9.4 to obtain the fecal occult blood collecting solution.

Example 3:

adding 70mL of purified water into a container, then sequentially adding 1.2g of disodium hydrogen phosphate, 0.5g of casein, 2g of 3-hydroxy-2, 2' iminodisuccinic acid tetrasodium (HIDS), 2g of sodium tripolyphosphate, 3g of polyethylene glycol, 0.5g of Proclin 150 and 0.5g of Proclin300, uniformly mixing, then fixing the volume to 100mL, and adjusting the pH to 9.5 to obtain the fecal occult blood collecting solution.

Example 4:

adding 70mL of purified water into a container, then sequentially adding 0.8g of borax, 1g of calf serum albumin, 1g of Ethylene Diamine Tetraacetic Acid (EDTA), 2g of sodium tripolyphosphate, 2.6g of polyethylene glycol, 0.25g of Proclin 150 and 0.25g of Proclin300, uniformly mixing, fixing the volume to 100mL, and adjusting the pH value to 9.5 to obtain the fecal occult blood collecting fluid.

Example 5:

adding 70mL of purified water into a container, then sequentially adding 1g of Tris (hydroxymethyl) aminomethane, 0.2g of calf serum albumin, 2g of 3-hydroxy-2, 2' tetrasodium iminodisuccinate (HIDS), 1.5g of Ethylene Diamine Tetraacetic Acid (EDTA), 2.8g of polyethylene glycol, 0.03g of Proclin 150 and 0.03g of Proclin300, uniformly mixing, fixing the volume to 100mL, and adjusting the pH to 9.4 to obtain the fecal occult blood collecting solution.

Application example 1:

taking several types of excrement occult blood excrement collecting liquid which are conventionally used in the market at present, wherein the collecting liquid A is the collecting liquid provided by company A, and the formula comprises EDTA and sodium chloride, wherein the concentrations are 3-10 g/L and 5-20 g/L respectively; the collection liquid B is provided by company B, and the formula comprises HEPES, calf serum albumin, sodium chloride, sucrose and a preservative, wherein the concentrations are 0.5-5 g/L, 1-10 g/L, 5-20 g/L, 5-30 g/L and 0.2-1 g/L respectively.

50mg of fresh fecal occult blood positive stool samples are respectively added into the three fecal collecting solutions of 2ml, and are placed at normal temperature, 2-8 ℃ and-20 ℃ for storage, and the performance is evaluated in 0 day, 3 days, 7 days, 10 days, 15 days, 30 days, 2 months, 3 months, 4 months, 5 months, 6 months, 9 months and 12 months, which are shown in Table 1.

TABLE 1 results of stool preservation evaluation using different collection fluids according to the application examples

Note: "+" indicates a positive result and "-" indicates a negative result.

The above results show that the collection prepared in example 1 can be stored at ambient temperature for 7 days, at 2-8 ℃ for 30 days and at-20 ℃ for 6 months; the stabilizing effect of the collected liquid in comparative example A, the collected liquid in comparative example B, and the hemoglobin in comparative example 1 and comparative example 2 is obviously inferior to that of the collected liquid prepared in example 1.

The above embodiments are not intended to limit the present invention, and the present invention is not limited to the above embodiments, and all embodiments are within the scope of the present invention as long as the requirements of the present invention are met.

Claims

kinds of collecting liquid for protecting and stabilizing hemoglobin in feces is characterized by comprising a chelating agent, a preservative, a sealing agent, a buffering agent and polyethylene glycol, wherein the concentration of each component is 25-45 g/L, 0.2-1 g/L, 1-10 g/L, 6-12 g/L and 25-30 g/L respectively, and the pH value is 9.4-9.5;

the chelating agent is 3-hydroxy-2, 2^/Tetrasodium iminodisuccinate (HIDS) and Ethylene Diamine Tetraacetic Acid (EDTA);

the preservative is a mixture of Proclin 150 and Proclin 300;

the sealant is kinds of calf serum albumin and casein;

the buffer is of Tris (hydroxymethyl) aminomethane, disodium hydrogen phosphate and borax.
2. A collection liquid for the protection and stabilization of hemoglobin in feces according to claim 1, which is composed of 3-hydroxy-2, 2/iminodisuccinic acid tetrasodium HIDS, EDTA, Proclin300, Proclin 150, Tris, calf serum albumin, polyethylene glycol, purified water, and the concentrations of HIDS, EDTA, Proclin300, Proclin 150, Tris, calf serum albumin, polyethylene glycol are 35g/L, 10g/L, 0.2g/L, 6.05g/L, 10g/L, 25g/L, respectively.
3. Use of a collection of claim 1 for the protection and stabilization of hemoglobin in stool for pretreatment prior to stool testing.
4. The use of claim 3, wherein fresh feces is mixed with the collection liquid of claim 1 in a ratio of 50mg to 2ml, and then the mixture is stored at room temperature, 2-8 ℃ or-20 ℃.
5. Use according to claim 4, characterized in that the storage time at ambient temperature is up to 7 days; the preservation time at 2-8 ℃ is as long as 1 month; the storage time at-20 deg.C is up to 6 months.