CN113341119A - Method for indirectly detecting lactose intolerance - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 43
- 201000010538 Lactose Intolerance Diseases 0.000 title claims abstract description 30
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 95
- 239000008103 glucose Substances 0.000 claims abstract description 95
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims abstract description 91
- 239000008101 lactose Substances 0.000 claims abstract description 89
- 238000001514 detection method Methods 0.000 claims abstract description 74
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 59
- 230000002550 fecal effect Effects 0.000 claims abstract description 36
- 102100026189 Beta-galactosidase Human genes 0.000 claims abstract description 29
- 108010005774 beta-Galactosidase Proteins 0.000 claims abstract description 29
- 108010059881 Lactase Proteins 0.000 claims abstract description 19
- 229940116108 lactase Drugs 0.000 claims abstract description 19
- 238000007865 diluting Methods 0.000 claims abstract description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 40
- 239000008213 purified water Substances 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 229960000583 acetic acid Drugs 0.000 claims description 20
- 239000012362 glacial acetic acid Substances 0.000 claims description 20
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 241000894006 Bacteria Species 0.000 claims description 3
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 3
- 238000003556 assay Methods 0.000 claims description 3
- 230000007062 hydrolysis Effects 0.000 abstract description 6
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 6
- 210000003608 fece Anatomy 0.000 description 12
- 239000007789 gas Substances 0.000 description 6
- 150000002772 monosaccharides Chemical class 0.000 description 6
- 230000015556 catabolic process Effects 0.000 description 4
- 210000001072 colon Anatomy 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 230000013872 defecation Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010023648 Lactase deficiency Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
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Abstract
The invention provides a method for indirectly detecting lactose intolerance, which comprises the following steps: obtaining a fecal sample, and obtaining the glucose content of the fecal sample; diluting a fecal sample to obtain a detection sample, and adding a reagent into the detection sample to obtain the glucose content in the detection sample; and subtracting the glucose content in the detection sample from the glucose content in the fecal sample to obtain the lactose content. Detecting lactose in the excrement by an indirect method, firstly detecting glucose in the excrement, then adding lactase to hydrolyze the lactose, and then detecting the glucose content after hydrolysis, wherein the lactose content is obtained by subtracting the glucose content from the glucose content; the method provided by the application can be used for quickly equally dividing one excrement sample and further simultaneously measuring the equally divided samples, so that the accuracy of lactose tolerance detection of one excrement sample is greatly improved; the method effectively reduces the situation that a sample detection result in the prior art has high probability of false positive.
Description
Technical Field
The invention relates to the technical field of lactose intolerance detection, in particular to a method for indirectly detecting lactose intolerance.
Background
Lactose intolerance of infants caused by small intestine lactase deficiency can prevent partial lactose from being completely absorbed, so that lactose directly reaches colon, and uncomfortable symptoms such as diarrhea and the like are induced; when a doctor diagnoses whether the infants have lactose intolerance, the doctor mainly relies on clinical experience and auxiliary detection at present;
the existing auxiliary detection of lactose intolerance of infants mainly comprises the following types:
1. the hydrogen breath test has problems that: the difficulty of collecting gas by infants;
2. the sugar tolerance test has problems that: the false positive rate is high, and the specificity is poor;
3. the biopsy of small intestinal mucosa has the following problems: invasive detection, requiring puncture;
4. the gene diagnosis has the following problems: the method is complicated, time-consuming and high in charging rate, and the false positive rate is up to 30%;
5. the problems of stool pH or reducing substance measurement are that: simple and easy to operate, and high in false positive rate;
6. the urine galactose detection method has the following problems: the detection environment is extremely harsh, so that the detection result has very large error;
among the above-mentioned detection means, the most recommended detection of infants is the measurement of fecal pH or reducing substances, but because of the high probability of false positive, a detection method capable of improving the detection accuracy is lacking.
Disclosure of Invention
The invention provides a method for indirectly detecting lactose intolerance, which is used for achieving the purposes of improving the detection efficiency and improving the detection accuracy when the lactose intolerance of infants is detected.
The invention provides a method for indirectly detecting lactose intolerance, which comprises the following steps:
step 1, obtaining a fecal sample, and obtaining the glucose content of the fecal sample;
and 3, subtracting the glucose content in the detected sample from the glucose content in the excrement sample to obtain the lactose content.
Preferably, the step 2 further includes:
diluting the excrement sample, and filtering bacteria on the diluted excrement sample by using a filter to obtain a detection sample.
Preferably, the step 2 includes:
injecting a lactase reagent into the detection sample, so that the detection sample is hydrolyzed into lactose, and then, the lactose is hydrolyzed into glucose; and after the content of the lactose obtained in the detection sample is detected, the glucose content of the detection sample is obtained.
Preferably, the method further comprises equally dividing the stool sample into a plurality of sub-stool samples, reserving one or more sub-stool samples, and obtaining the glucose content of the sub-stool samples;
diluting the rest of the subsidiary excrement samples into subsidiary detection samples to obtain the glucose content of the subsidiary detection samples;
subtracting the glucose content of the sub-assay sample from the glucose content of the sub-fecal sample; the lactose content is obtained.
Preferably, the lactose content is: 1mol lactose is hydrolyzed to 1mol glucose.
Preferably, fecal samples are obtained for a plurality of time periods, and the glucose content of the fecal samples for each time period and the glucose content of the test sample obtained from the fecal sample are obtained;
subtracting the glucose content of the detection sample in the corresponding time period from the glucose content of the fecal sample in each time period; lactose content was obtained for various time periods.
Preferably, the reagents include reagent 1, reagent 2 and reagent 3;
the reagent 1 is a sulfate solvent, and the preparation amount is 100-1200 ml;
the reagent 2 is a glacial acetic acid solvent, and the preparation amount is 100-1200 ml;
the reagent 3 is beta-galactosidase and glacial acetic acid solvent, and the preparation amount is 100-1200 ml.
Preferably, the reagent 1, the reagent 2 and the reagent 3 are respectively injected into the diluted sub-detection samples in sequence, and the glucose content of the plurality of sub-detection samples is obtained;
and subtracting the glucose content of each subsidiary detection sample from the glucose content of the subsidiary fecal samples to obtain a plurality of lactose contents, and comparing the lactose contents with a preset value to obtain a lactose tolerance detection result of the fecal samples.
Preferably, the reagent 1 comprises: 4-6.5g of disodium hydrogen phosphate with the concentration of 0.50-0.65%; 0.35 to 0.68g of disodium hydrogen phosphate with the concentration of 0.040 to 0.068 percent; 7.0-10.5g of sodium chloride with the concentration of 0.75-1.20%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml;
the reagent 2 comprises: 36-48g of glacial acetic acid with the concentration of 3.65% -4.82%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml;
the reagent 3 comprises: 4.0-6.5g of beta-galactosidase with the concentration of 0.42% -0.65%; 36-48g of glacial acetic acid with the concentration of 3.65% -4.82%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml.
Preferably, the reagent 1 comprises: 5.8g of disodium hydrogen phosphate with the concentration of 0.58 percent; 0.59g of disodium hydrogen phosphate with the concentration of 0.059 percent; 9.0g of sodium chloride with the concentration of 0.9 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water;
the reagent 2 comprises: 40.0g of glacial acetic acid with the concentration of 4.0 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water;
the reagent 3 comprises: 5.0g of beta-galactosidase with the concentration of 0.5 percent; 40.0g of glacial acetic acid with the concentration of 4.0 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water.
According to the invention, lactose in excrement is detected by an indirect method, glucose in the excrement is detected firstly, then lactase is added to hydrolyze lactose, and then the glucose content after hydrolysis is detected, wherein the lactose content is obtained by subtracting the glucose content from the glucose content; wherein, the lactose content is calculated according to the molar mass, 1mol of lactose is hydrolyzed to generate 1mol of glucose, so as to obtain the lactose content in the infant feces, and further judge whether the infant has lactose intolerance or not according to the lactose content;
according to the method, the method can be simple and effective, and compared with the prior art, the method provided by the application can be used for quickly equally dividing one excrement sample and further simultaneously measuring the equally divided samples, so that the accuracy of lactose tolerance detection of one excrement sample is greatly improved; the method effectively reduces the situation that a sample detection result in the prior art has high probability of false positive.
Furthermore, the method is used for diluting and filtering the excrement sample, adding lactase to hydrolyze lactose into glucose, detecting the glucose, calculating the content of residual lactose in the excrement according to the result, and providing auxiliary diagnosis information for doctors by combining daily defecation amount and daily lactose intake amount.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and drawings.
The technical solution of the present invention is further described in detail by the accompanying drawings and embodiments.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a flow chart of the present invention;
FIG. 2 is a statistical value of the results of the detection of the existing single stool sample;
FIG. 3 is a statistical result of the testing of various types of stool samples using the present invention.
Detailed Description
The preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.
According to the embodiments of the present invention as shown in FIGS. 1-3, a method for indirectly detecting lactose intolerance is provided, which comprises the following steps:
step 1, obtaining a fecal sample, and obtaining the glucose content of the fecal sample;
and 3, subtracting the glucose content in the detected sample from the glucose content in the excrement sample to obtain the lactose content.
In the step 2, the method further comprises: diluting the excrement sample, and filtering bacteria on the diluted excrement sample by using a filter to obtain a detection sample. Further, injecting a lactase reagent into the detection sample to hydrolyze the detection sample into lactose, and then hydrolyzing the lactose into glucose; and after the content of the lactose obtained in the detection sample is detected, the glucose content of the detection sample is obtained.
Equally dividing the excrement sample into a plurality of sub excrement samples, reserving one or more sub excrement samples, and obtaining the glucose content of the sub excrement samples;
diluting the rest of the subsidiary excrement samples into subsidiary detection samples to obtain the glucose content of the subsidiary detection samples;
subtracting the glucose content of the sub-assay sample from the glucose content of the sub-fecal sample; the lactose content is obtained.
The lactose content is as follows: 1mol lactose is hydrolyzed to 1mol glucose.
Obtaining fecal samples of a plurality of time periods, and obtaining the glucose content of the fecal samples of each time period and the glucose content of a test sample obtained from the fecal samples;
subtracting the glucose content of the detection sample in the corresponding time period from the glucose content of the fecal sample in each time period; lactose content was obtained for various time periods.
According to the invention, lactose in excrement is detected by an indirect method, glucose in the excrement is detected firstly, then lactase is added to hydrolyze lactose, and then the glucose content after hydrolysis is detected, wherein the lactose content is obtained by subtracting the glucose content from the glucose content; wherein, the lactose content is calculated according to the molar mass, 1mol of lactose is hydrolyzed to generate 1mol of glucose, so as to obtain the lactose content in the infant feces, and further judge whether the infant has lactose intolerance or not according to the lactose content;
according to the method, the method can be simple and effective, and compared with the prior art, the method provided by the application can be used for quickly equally dividing one excrement sample and further simultaneously measuring the equally divided samples, so that the accuracy of lactose tolerance detection of one excrement sample is greatly improved; the method effectively reduces the situation that the detection result of one sample has high probability of false positive in the prior art.
Furthermore, the method is used for diluting and filtering the excrement sample, adding lactase to hydrolyze lactose into glucose, detecting the glucose, calculating the content of residual lactose in the excrement according to the result, and providing auxiliary diagnosis information for doctors by combining daily defecation amount and daily lactose intake amount.
In one embodiment, the reagents include reagent 1, reagent 2, and reagent 3;
the reagent 1 is a sulfate solvent, and the preparation amount is 100-1200 ml;
the reagent 2 is a glacial acetic acid solvent, and the preparation amount is 100-1200 ml;
the reagent 3 is beta-galactosidase and glacial acetic acid solvent, and the preparation amount is 100-1200 ml.
Sequentially injecting the reagent 1, the reagent 2 and the reagent 3 into the diluted sub-detection samples respectively, and obtaining the glucose content of the plurality of sub-detection samples;
and subtracting the glucose content of each subsidiary detection sample from the glucose content of the subsidiary fecal samples to obtain a plurality of lactose contents, and comparing the lactose contents with a preset value to obtain a lactose tolerance detection result of the fecal samples.
The reagent 1 comprises: 4-6.5g of disodium hydrogen phosphate with the concentration of 0.50-0.65%; 0.35 to 0.68g of disodium hydrogen phosphate with the concentration of 0.040 to 0.068 percent; 7.0-10.5g of sodium chloride with the concentration of 0.75-1.20%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml;
the reagent 2 comprises: 36-48g of glacial acetic acid with the concentration of 3.65% -4.82%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml;
the reagent 3 comprises: 4.0-6.5g of beta-galactosidase with the concentration of 0.42% -0.65%; 36-48g of glacial acetic acid with the concentration of 3.65% -4.82%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml.
The reagent 1 comprises: 5.8g of disodium hydrogen phosphate with the concentration of 0.58 percent; 0.59g of disodium hydrogen phosphate with the concentration of 0.059 percent; 9.0g of sodium chloride with the concentration of 0.9 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water;
the reagent 2 comprises: 40.0g of glacial acetic acid with the concentration of 4.0 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water;
the reagent 3 comprises: 5.0g of beta-galactosidase with the concentration of 0.5 percent; 40.0g of glacial acetic acid with the concentration of 4.0 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water.
In the embodiment, lactose in excrement is detected by an indirect method, glucose in the excrement is detected firstly, then lactase is added to hydrolyze the lactose, and then the glucose content after hydrolysis is detected, wherein the lactose content is obtained by subtracting the glucose content from the lactose content; wherein, the lactose content is calculated according to the molar mass, 1mol of lactose is hydrolyzed to generate 1mol of glucose, so as to obtain the lactose content in the infant feces, and further judge whether the infant has lactose intolerance or not according to the lactose content;
according to the method, the method can be simple and effective, and compared with the prior art, the method provided by the application can be used for quickly equally dividing one excrement sample and further simultaneously measuring the equally divided samples, so that the accuracy of lactose tolerance detection of one excrement sample is greatly improved; the method effectively reduces the situation that the detection result of one sample has high probability of false positive in the prior art.
Furthermore, the method is used for diluting and filtering the excrement sample, adding lactase to hydrolyze lactose into glucose, detecting the glucose, calculating the content of residual lactose in the excrement according to the result, and providing auxiliary diagnosis information for doctors by combining daily defecation amount and daily lactose intake amount.
The three reagents are used for respectively carrying out lactose hydrolysis on a plurality of sub-detection samples so as to obtain a plurality of detection results, and the average value of the plurality of detection results is obtained so as to obtain the most accurate detection result, specifically, the reagent 1: the main components are as follows: phosphate, solvent: purified water containing Proclin-300; reagent 2: the main components are as follows: glacial acetic acid, solvent: purified water containing Proclin-300; reagent 3: the main components are as follows: beta-galactosidase, glacial acetic acid, solvent: purified water containing Proclin-300.
Further, the present invention is used to perform experimental comparison between lactose tolerance and lactose intolerance, thereby analyzing the possible existence of the following 4 cases, further judging the lactose tolerance state of the subject according to the following four cases, and evaluating whether medical intervention is needed according to the lactose tolerance state. In particular, as shown in figures 1-3,
sample analysis was performed on the normal and lactose intolerant groups, further dividing the normal and lactose intolerant groups into four phases, as shown in table 1:
TABLE 1 sample Collection information Table for Normal group and lactose intolerant group
The test was performed in four types of test groups in table 1, wherein, of the 0-6 month-old type, the ages of 15 normal group samples and 15 lactose intolerance group samples were specifically 0-5 months old; in the 7-12 month age types, the age of 17 normal group samples and 15 lactose intolerance group samples is specifically 6-12 months; wherein the normal group is abbreviated Nor and the lactose intolerant group is abbreviated LI.
Further, fig. 2 shows the test result of the prior art for the stool sample alone: wherein, the number of samples of the normal group is 32, and the number of samples of the lactose intolerance group is 30; the average accuracy of the total amount of glucose was 5%; the average accuracy of the hydrolyzed glucose was 6%; the average accuracy of fecal glucose was not counted; the average accuracy of the lactose degradation rate was 18%.
FIG. 3 shows the test structure by the method of the present invention, and the sample numbers of the normal group and the lactose intolerant group are shown in Table 1,
0-5 age group: the average accuracy of the total amount of glucose was 69%; the average accuracy of the hydrolyzed glucose was 68%; the average accuracy of fecal glucose is 36%; the average accuracy of lactose degradation was 88%.
In 6-12 months of age: the average accuracy of fecal glucose is 22%; the average accuracy of lactose degradation rate was 16%.
According to the number of samples, the following four judgment standards are counted and obtained, and further, reference is made according to the following four judgment standards of lactose intolerance, so that in the process of detecting other lactose intolerance, the judgment of which type the lactose intolerance of the children patient belongs to can be carried out based on the standards;
the first type is that the activity of flora lactase is strong, lactose entering colon is rapidly hydrolyzed into monosaccharide (less lactose residue in feces) by the flora lactase, but the flora is slowly utilized and is discharged out of body, less acid and gas are generated at the moment, the concentration of glucose in feces is detected to be higher, the pH of feces is normal, the gas production of a fermented lactose culture medium is less, and the symptoms of children patients are not serious;
the second type, the activity of the flora lactase is strong, lactose entering colon is rapidly hydrolyzed into monosaccharide (less lactose residue in feces) by the flora lactase, the flora rapidly utilizes the monosaccharide for fermentation to generate a large amount of acid and gas, the concentration of glucose in feces is detected to be low, the pH of the feces is low, the fermented lactose culture medium generates more gas, and the symptoms of children patients are serious;
in the third type, the flora has less lactase or poor activity, lactose entering colon is slowly hydrolyzed into monosaccharide (feces have more lactose residues), the utilization rate of the monosaccharide by the flora is limited due to slow hydrolysis, the monosaccharide is probably similar to that of the first type, the concentration of glucose in feces is detected to be higher, the pH of feces is normal, the fermented lactose culture medium has less gas production, and a patient has diarrhea to a certain degree due to a large amount of lactose;
in the fourth type, normal children with endogenous lactase or partial lactase have less fecal glucose and less glucose released by continued degradation, and can be judged to be a group with normal or relatively normal lactase by combining the milk intake of the previous two days.
In order to distinguish the first type from the third type, the lactase is introduced, the glucose content is further detected, and a fourth judgment result can be obtained according to the symptoms of the infants, wherein the detection result is accurate and reliable.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (10)
1. A method for indirectly detecting lactose intolerance, comprising the steps of:
step 1, obtaining a fecal sample, and obtaining the glucose content of the fecal sample;
step 2, diluting the fecal sample into a detection sample, and adding a reagent into the detection sample to obtain the glucose content in the detection sample;
and 3, subtracting the glucose content in the detected sample from the glucose content in the excrement sample to obtain the lactose content.
2. The method according to claim 1, wherein said step 2 further comprises:
diluting the excrement sample, and filtering bacteria on the diluted excrement sample by using a filter to obtain a detection sample.
3. The method according to claim 2, wherein said step 2 comprises:
injecting a lactase reagent into the detection sample, so that the detection sample is hydrolyzed into lactose, and then, the lactose is hydrolyzed into glucose; and after the content of the lactose obtained in the detection sample is detected, the glucose content of the detection sample is obtained.
4. The method according to claim 3, further comprising dividing the stool sample into a plurality of sub-stool samples, collecting one or more of the sub-stool samples, and obtaining the glucose content of the sub-stool samples;
diluting the rest of the subsidiary excrement samples into subsidiary detection samples to obtain the glucose content of the subsidiary detection samples;
subtracting the glucose content of the sub-assay sample from the glucose content of the sub-fecal sample; the lactose content is obtained.
5. The method of claim 1, wherein the lactose content is: 1mol lactose is hydrolyzed to 1mol glucose.
6. The method of claim 1, wherein fecal samples are obtained for a plurality of time periods and the glucose content of the fecal samples for each time period and the glucose content of the test sample obtained from the fecal sample are obtained;
subtracting the glucose content of the detection sample in the corresponding time period from the glucose content of the fecal sample in each time period; lactose content was obtained for various time periods.
7. The method of claim 4, wherein said reagents comprise reagent 1, reagent 2, and reagent 3;
the reagent 1 is a sulfate solvent, and the preparation amount is 100-1200 ml;
the reagent 2 is a glacial acetic acid solvent, and the preparation amount is 100-1200 ml;
the reagent 3 is beta-galactosidase and glacial acetic acid solvent, and the preparation amount is 100-1200 ml.
8. The method according to claim 7, wherein reagent 1, reagent 2 and reagent 3 are sequentially injected into the diluted sub-samples, and the glucose content of the sub-samples is obtained;
and subtracting the glucose content of each subsidiary detection sample from the glucose content of the subsidiary fecal samples to obtain a plurality of lactose contents, and comparing the lactose contents with a preset value to obtain a lactose tolerance detection result of the fecal samples.
9. The method of claim 7, wherein said lactose intolerance is detected indirectly,
the reagent 1 comprises: 4-6.5g of disodium hydrogen phosphate with the concentration of 0.50-0.65%; 0.35 to 0.68g of disodium hydrogen phosphate with the concentration of 0.040 to 0.068 percent; 7.0-10.5g of sodium chloride with the concentration of 0.75-1.20%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml;
the reagent 2 comprises: 36-48g of glacial acetic acid with the concentration of 3.65% -4.82%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml;
the reagent 3 comprises: 4.0-6.5g of beta-galactosidase with the concentration of 0.42% -0.65%; 36-48g of glacial acetic acid with the concentration of 3.65% -4.82%; proclin 3000.8-1.3 ml with the concentration of 0.08% -0.15%; purified water 980-1000 ml.
10. The method of claim 9, wherein said detecting comprises detecting lactose intolerance,
the reagent 1 comprises: 5.8g of disodium hydrogen phosphate with the concentration of 0.58 percent; 0.59g of disodium hydrogen phosphate with the concentration of 0.059 percent; 9.0g of sodium chloride with the concentration of 0.9 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water;
the reagent 2 comprises: 40.0g of glacial acetic acid with the concentration of 4.0 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water;
the reagent 3 comprises: 5.0g of beta-galactosidase with the concentration of 0.5 percent; 40.0g of glacial acetic acid with the concentration of 4.0 percent; 3001.0 ml of Proclin with the concentration of 0.1 percent; 998ml of purified water.
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