Klebsiella pneumoniae content determination method and detection product

Abstract

The invention provides a Klebsiella pneumoniae content determination method and a detection product, and relates to the technical field of pathogen detection. The method for determining the content of the klebsiella pneumoniae is a method for determining the content of the klebsiella pneumoniae in a target sample by taking glucose-6-phosphate dehydrogenase as a specific recognition site. According to the application, the characteristic protein (glucose-6-phosphate dehydrogenase) on the Klebsiella pneumoniae capsular polysaccharide is specifically recognized by adopting the horse radish peroxidase-labeled antibody, then the chromogenic reaction is carried out, and the detection instrument is combined for detection, so that whether the to-be-detected sample contains the Klebsiella pneumoniae or not is analyzed, the detection time is shortened, the operation is simple, and the detection efficiency is high. The detection cost is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of pathogen detection, and in particular to a method for determining the content of Klebsiella pneumoniae and a detection product. Background Art

[0002] Klebsiella pneumoniae (KP) is a Gram-negative bacterium that is widely distributed in the natural environment, including soil, water, and animal intestines. It can cause a variety of infections in humans, including urinary tract infections, pneumonia, and sepsis. Since KP is resistant to multiple antibiotics, treatment becomes complicated. Therefore, rapid and accurate KP detection methods are essential to control its spread and treat infections.

[0003] Currently, the detection methods of KP include traditional bacterial culture, oxidation / reduction test, enzyme activity test and molecular biology test. The bacterial culture method involves culturing patient samples such as sputum, blood or urine on culture medium and isolating KP. This process is time-consuming and may take several days to more than ten days, delaying the diagnosis and treatment of patients. There is also a risk of sample contamination. Oxidation / reduction and enzyme activity tests identify KP based on the specific biochemical characteristics of bacteria. These methods are not only time-consuming and labor-intensive, but also have low specificity in the presence of other similar Gram-negative bacteria, which can easily lead to misjudgment.

[0004] Molecular biological detection methods, such as polymerase chain reaction (PCR) and gene sequencing, identify specific gene sequences of KP by detecting bacterial DNA in samples. Although these methods are highly accurate, the experimental process is complicated and difficult to operate. They require specialized equipment and professional training personnel, which increases the cost of analysis. In addition, these methods may face problems such as improper sample handling and insufficient equipment precision in actual operation, further affecting the accuracy and reliability of the test results.

[0005] Considering the defects of the existing technologies, it can be seen that the detection methods of KP generally have problems such as long time consumption, complicated operation, high cost and low specificity. The long waiting time of bacterial culture method may lead to delays in clinical treatment, and the low specificity of oxidation / reduction test and enzyme activity test may cause misdiagnosis. Although molecular biological detection is accurate, its high cost and complexity of operation limit its application in resource-limited environments. Therefore, it is particularly urgent to develop a fast, simple, low-cost and highly specific KP detection method, which is of great significance for improving the diagnostic efficiency and treatment success rate of KP infection.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] The first purpose of the present invention is to provide a method for determining the content of Klebsiella pneumoniae. The method for determining the content of Klebsiella pneumoniae has the advantages of wide application range, simple and rapid operation method, accurate results, etc.

[0008] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0009] In a first aspect, the present invention provides a method for determining the content of Klebsiella pneumoniae. The method for determining the content of Klebsiella pneumoniae is a method for determining the content of Klebsiella pneumoniae in a target sample using glucose-6-phosphate dehydrogenase as a specific recognition site.

[0010] In an optional embodiment, the method for determining the content of Klebsiella pneumoniae comprises:

[0011] Set up different experimental groups;

[0012] For different experimental groups, the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase-labeled antibody and reaction substrate are added, and incubated respectively to obtain incubated samples of each experimental group;

[0013] Detecting the incubated samples in each experimental group to obtain absorbance values;

[0014] A standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value is drawn, and the Klebsiella pneumoniae content in the target sample is calculated according to the standard curve.

[0015] In an optional embodiment, the different experimental groups respectively include:

[0016] Blank group, test sample group and standard group.

[0017] In an optional embodiment, the blank group contains a sterile solution;

[0018] The sample group to be tested contains a suspension of the target sample;

[0019] The standard set includes a glucose-6-phosphate dehydrogenase solution with a known concentration.

[0020] In an optional embodiment, the step of drawing a standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value according to the detection data, and calculating the Klebsiella pneumoniae content in the target sample according to the standard curve comprises:

[0021] Using each sample in the standard group, the standard curve is drawn with the concentration of glucose-6-phosphate dehydrogenase as the abscissa and the absorbance value as the ordinate;

[0022] The absorbance value of the target sample is brought into the standard curve to calculate the content of Klebsiella pneumoniae in the target sample.

[0023] In an optional embodiment, the incubated samples of each experimental group are tested to obtain the absorbance value, and the detection method used includes at least one of ultraviolet spectrophotometry and enzyme labeling method;

[0024] In an alternative embodiment, the detection wavelength is 450 nm.

[0025] In an optional embodiment, for different experimental groups, the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase-labeled antibody and reaction substrate are added and incubated separately, including:

[0026] A sterile solution is added to the blank group; a glucose-6-phosphate dehydrogenase solution is added to the standard group; and a suspension of the target sample is added to the test sample group;

[0027] Performing a first incubation on the blank group, the standard group and the test sample group;

[0028] After adding horseradish peroxidase-labeled antibody to the blank group, the standard group and the test sample group, a second incubation is performed;

[0029] Adding the reaction substrate to the blank group, the standard group and the sample group to be tested respectively for a third incubation, thus obtaining incubated samples of each experimental group;

[0030] In an optional embodiment, the reaction substrate is 3,3',5,5'-tetramethylbenzidine;

[0031] In an optional embodiment, the incubation temperatures of the first incubation, the second incubation and the third incubation are all: 37°C;

[0032] In an optional embodiment, the incubation time of the first incubation is 20 minutes;

[0033] In an optional embodiment, the incubation time of the second incubation is 30 minutes;

[0034] In an optional embodiment, the incubation time of the third incubation is 20 minutes.

[0035] In an optional embodiment, the dilution ratio of the horseradish peroxidase-labeled antibody is 1:30,000.

[0036] In an optional embodiment, in the standard group, the samples of glucose-6-phosphate dehydrogenase solution with different known concentrations have concentration values ​​that satisfy an orderly increasing arrangement of geometric progression from small to large;

[0037] In an optional embodiment, the number of samples containing glucose-6-phosphate dehydrogenase solutions with different concentrations in the standard set is at least 5.

[0038] In a second aspect, the present invention further provides a detection product, using the method for determining the content of Klebsiella pneumoniae as described in any one of the above embodiments;

[0039] The detection product includes at least one of a detection kit, a detection reagent and a detection device.

[0040] The present invention provides a method for determining the content of Klebsiella pneumoniae and a detection product, which is a method for determining the content of Klebsiella pneumoniae in a target sample by using glucose-6-phosphate dehydrogenase as a specific recognition site. The method for identifying Klebsiella pneumoniae based on capsular polysaccharide provided by the present application uses a horseradish peroxidase-labeled antibody to specifically identify a characteristic protein (glucose-6-phosphate dehydrogenase) on the capsular polysaccharide of Klebsiella pneumoniae, and then performs a color reaction, and performs detection in combination with a detection instrument, thereby analyzing whether the sample to be tested contains Klebsiella pneumoniae, reducing detection time, simplifying operation, and reducing detection costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0042] Figure 1 Schematic diagram of the process of the method for determining the content of Klebsiella pneumoniae in the examples of the present application. DETAILED DESCRIPTION

[0043] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be obtained commercially.

[0044] The present application provides a method for determining the content of Klebsiella pneumoniae in a target sample by using glucose-6-phosphate dehydrogenase as a specific recognition site.

[0045] It should be noted that Klebsiella pneumoniae (KP) is a Gram-negative bacillus belonging to the genus Klebsiella of the family Enterobacteriaceae. It is widely present in the natural environment, including soil, water, and the intestinal tract and respiratory tract of humans and animals. KP is an opportunistic pathogen that usually causes infection when the host's immune system is dysfunctional, leading to serious diseases such as pneumonia, sepsis, and meningitis. The bacterium has a variety of pathogenic factors, such as capsular polysaccharides (K antigens), which can evade the host's immune defense and increase the risk of infection.

[0046] In the present application example, glucose-6-phosphate dehydrogenase (G6PD) is used as a specific recognition site of Klebsiella pneumoniae for detection, which has many advantages. First, G6PD is highly conserved in KP, and capsular polysaccharide is an important virulence factor of KP, and G6PD is a characteristic protein of capsular polysaccharide, which makes it an ideal target and can be used to develop a highly specific detection method. Secondly, G6PD is closely related to the virulence of KP, and its yield is usually the main criterion for measuring the virulence characteristics of strains. Therefore, detecting G6PD can not only identify KP, but also evaluate its potential pathogenicity. In addition, the detection method based on G6PD can be designed to be simple and rapid, reduce the complexity and time of operation, and reduce the detection cost at the same time, which is of great significance for the rapid clinical diagnosis and treatment of KP infection.

[0047] In conclusion, the use of G6PD as a specific recognition site for the detection of Klebsiella pneumoniae can provide a rapid, simple, low-cost and highly specific detection method, which has significant advantages in improving the diagnostic efficiency and treatment success rate of KP infection.

[0048] For further reference, Figure 1 , the method for determining the content of Klebsiella pneumoniae comprises:

[0049] Step S1, setting different experimental groups;

[0050] In one embodiment, the different experimental groups respectively include:

[0051] Blank group, test sample group and standard group.

[0052] In the above steps, first, you need to prepare multiple experimental groups for control and standard curve drawing. These experimental groups usually include:

[0053] Blank group: does not contain target samples and G6PD, used to determine the background signal of the experiment.

[0054] Standard group: Contains G6PD solutions with known different concentrations, used to draw the standard curve.

[0055] Test sample group: contains target samples to be tested, used to determine the content of Klebsiella pneumoniae therein.

[0056] Step S2, for different experimental groups, adding the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase-labeled antibody and reaction substrate, and incubating them respectively to obtain incubated samples of each experimental group.

[0057] As described above, for each experimental group, target samples and G6PD solutions of different concentrations are added. Horseradish peroxidase (HRP)-labeled antibodies are added, which can specifically recognize G6PD. A reaction substrate is added, which can be 3,3',5,5'-tetramethylbenzidine (TMB), which will produce a color change under the catalysis of HRP. The samples of each experimental group are incubated at a suitable temperature to allow sufficient reaction between the antibody and G6PD.

[0058] Step S3, testing the incubated samples in each experimental group to obtain absorbance values;

[0059] As described above, after the incubation is completed, the samples of each experimental group are tested using an ELISA reader or other photometer equipment. The absorbance values ​​of each sample are measured at a specific wavelength, and these values ​​are proportional to the concentration of G6PD in the sample.

[0060] Step S4, drawing a standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value according to the absorbance value, and calculating the Klebsiella pneumoniae content in the target sample according to the standard curve.

[0061] The standard curve is drawn using the absorbance values ​​of the standard group and the known G6PD concentration. This curve can reflect the absorbance values ​​generated by different concentrations of G6PD. By comparing the absorbance values ​​of the sample group with the standard curve, the content of Klebsiella pneumoniae in the sample can be calculated.

[0062] In summary, the advantage of the above method is its high specificity, because G6PD is a highly conserved antigen in Klebsiella pneumoniae, which can accurately identify and measure Klebsiella pneumoniae. In addition, this method is easy to operate, fast in detection, and relatively low in cost, which is suitable for rapid detection needs in clinical and research. Through this method, Klebsiella pneumoniae infection can be effectively monitored and controlled, which helps to take timely treatment measures.

[0063] Furthermore, the blank group contains a sterile solution;

[0064] The sample group to be tested contains a suspension of the target sample;

[0065] The standard set includes a glucose-6-phosphate dehydrogenase solution with a known concentration.

[0066] Furthermore, the step S4, drawing a standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value according to the detection data, and calculating the Klebsiella pneumoniae content in the target sample according to the standard curve, includes:

[0067] Step S41, using each sample in the standard group, with the concentration of glucose-6-phosphate dehydrogenase as the abscissa and the absorbance value as the ordinate, to draw the standard curve;

[0068] This step is used to establish a mathematical model to link the concentration of glucose-6-phosphate dehydrogenase (G6PD) in the standard group with its corresponding absorbance value. Specifically, the concentration of glucose-6-phosphate dehydrogenase is used as the horizontal coordinate (x-axis), and the absorbance value of each standard group sample is used as the vertical coordinate (y-axis). These data points are plotted on a graph, and linear regression or other appropriate statistical methods are usually used to fit these data points to form a standard curve. The obtained standard curve is usually linear, but it may also present a nonlinear relationship based on experimental data. This curve can reflect the absorbance values ​​produced by different concentrations of G6PD.

[0069] Step S42, bringing the absorbance value of the target sample into the standard curve to calculate the Klebsiella pneumoniae content in the target sample.

[0070] Above, in this step, it is used to utilize the standard curve to quantitatively analyze the content of Klebsiella pneumoniae in the target sample. Determine the absorbance value of the target sample, then bring the absorbance value of the target sample into the equation of the standard curve, and calculate the corresponding G6PD concentration. Since G6PD is a specific recognition site of Klebsiella pneumoniae, its concentration is proportional to the content of Klebsiella pneumoniae, so the content of Klebsiella pneumoniae in the target sample can be inferred by the concentration of G6PD. The relative quantitative result of Klebsiella pneumoniae in the target sample can be obtained by this step.

[0071] Furthermore, in step S3, the incubated samples of each experimental group are tested to obtain the absorbance value, and the detection method used includes at least one of ultraviolet spectrophotometry and enzyme labeling method;

[0072] Furthermore, the detection wavelength is 450 nm.

[0073] As mentioned above, UV spectrophotometry is an analytical technique based on UV-visible spectroscopy, which quantitatively analyzes the concentration of a sample by measuring the absorbance of a substance at a specific wavelength. In ELISA, UV spectrophotometry is usually used to measure the absorbance of a colored product produced after an enzyme-labeled reaction, thereby obtaining an absorbance value.

[0074] As mentioned above, the enzyme-linked immunosorbent assay (ELISA) is an immunological detection method that uses enzyme-labeled antibodies to bind to target antigens and quantitatively analyze target antigens by producing color changes in the enzyme-catalyzed substrate.

[0075] Alternatively, samples can be analyzed by measuring absorbance at specific wavelengths using high performance liquid chromatography or continuous flow analyzer coupled with a UV-visible detector.

[0076] As mentioned above, in the ultraviolet spectrophotometry, an ultraviolet spectrophotometer can be used; in the enzyme labeling method, an enzyme labeler can be used for detection.

[0077] In addition, according to different samples to be tested, a suitable detection carrier can be selected, for example, an ELISA plate or other carriers capable of adsorbing glucose-6-phosphate dehydrogenase antibodies, such detection containers are small in size and easy to operate.

[0078] Furthermore, the step S2, for different experimental groups, adds the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase labeled antibody and reaction substrate, and incubates them separately, including:

[0079] Step S21, adding a sterile solution to the blank group; adding a glucose-6-phosphate dehydrogenase solution to the standard group; and adding a suspension of the target sample to the test sample group;

[0080] Step S22, performing a first incubation on the blank group, the standard group and the test sample group.

[0081] In the above, in the first incubation, corresponding solutions are added to the blank group, the standard group and the sample group to be tested, which are respectively:

[0082] (1) Blank group: sterile solution; used as negative control

[0083] (2) Standard group: glucose-6-phosphate dehydrogenase solution; known concentration;

[0084] (3) Test sample group: suspension of target sample.

[0085] As mentioned above, this step is used to attach the antigen (glucose-6-phosphate dehydrogenase) in the target sample to the solid surface. Taking the ELISA method as an example, the solid surface may be an ELISA plate pre-coated with a capture antibody against the antigen. In this step, the antigen in the sample or standard can bind to the capture antibody on the solid surface to form an antigen-antibody complex.

[0086] Step S23, adding horseradish peroxidase-labeled antibody to the blank group, the standard group and the test sample group, respectively, and then performing a second incubation.

[0087] In the above, the detection antibody labeled with horseradish peroxidase (HRP) is bound to the antigen-antibody complex already bound to the solid surface. Specifically, after the first incubation, the detection antibody labeled with HRP can be added to each group, and the second incubation can be performed at 37°C. The detection antibody labeled with HRP will specifically bind to the antigen-antibody complex, preparing for the subsequent color development reaction. This step enhances the specificity of the detection because it involves the binding of two specific antibodies.

[0088] Step S24, adding the reaction substrate to the blank group, the standard group and the test sample group respectively for a third incubation, so as to obtain incubated samples of each experimental group;

[0089] As described above, a reaction substrate is added to catalyze a detectable color change. After the second incubation, a reaction substrate (such as 3,3',5,5'-tetramethylbenzidine, TMB) is added to each group and a third incubation is performed at 37°C. Horseradish peroxidase catalyzes the oxidation of the substrate TMB on the HRP-labeled detection antibody, resulting in a color change. This color change is proportional to the amount of antigen in the sample, and the amount of antigen can be quantitatively analyzed by measuring the absorbance.

[0090] Further, the reaction substrate is 3,3',5,5'-tetramethylbenzidine;

[0091] Furthermore, the incubation temperatures of the first incubation, the second incubation and the third incubation are all: 37°C;

[0092] Furthermore, the incubation time of the first incubation is 20 minutes;

[0093] Furthermore, the incubation time of the second incubation is 30 minutes;

[0094] Furthermore, the incubation time of the third incubation is 20 minutes.

[0095] As mentioned above, the duration and temperature of each incubation are optimized to ensure sufficient binding and reaction between the reactants while reducing non-specific binding. These steps are performed at 37°C because this temperature can accelerate the reaction while reducing non-specific binding. By precisely controlling the incubation time and temperature, the specificity and sensitivity of the ELISA experiment can be ensured, thereby obtaining accurate and reliable results.

[0096] Furthermore, the dilution ratio of the horseradish peroxidase-labeled antibody is 1:30000.

[0097] Furthermore, in the standard sample group, the samples of glucose-6-phosphate dehydrogenase solution with different concentrations are known, and their concentration values ​​satisfy the orderly increasing arrangement of geometric progression from small to large.

[0098] As mentioned above, setting the standard group with increasing concentrations in a geometric progression can ensure that evenly distributed data points are obtained in the entire measurement range, thereby drawing a smooth and accurate standard curve. Specifically, a concentration range can be determined first, which should cover a range from a lower limit far below the expected target molecule concentration in the sample to an upper limit far above the expected target molecule concentration in the sample.

[0099] A common ratio (usually greater than 1) can then be chosen to ensure that the concentration of each standard is a fixed multiple of the previous standard concentration. For example, if the lowest concentration is set to 1U / mL and the common ratio is 2, then the subsequent standard concentrations can be set to 2U / mL, 4U / mL, 8U / mL, etc.

[0100] As mentioned above, the geometric progression can ensure good linear response in both low and high concentration regions, which helps to improve the accuracy and reliability of the standard curve. This setting helps to capture potential nonlinear responses in different concentration regions, thereby more accurately estimating the concentration of the target molecule in the sample.

[0101] Furthermore, the number of samples containing glucose-6-phosphate dehydrogenase solutions of different concentrations in the standard sample group is at least 5.

[0102] As mentioned above, using at least 5 standard samples of different concentrations (such as 5, 6, 7, 9, etc.) can provide enough data points to draw a reliable standard curve, thereby improving the accuracy of quantitative analysis. Depending on the needs of the experimental design, 5 or more standard samples of different concentrations can be selected. These standard samples should evenly cover the expected measurement range to ensure that there are enough data points in the key areas.

[0103] As mentioned above, increasing the number of standard samples can improve the stability and reliability of the standard curve, especially in areas with large concentration changes. Multiple data points help identify and correct possible nonlinear trends, thereby improving accuracy across the entire measurement range. At least 5 standard samples can provide enough information to evaluate the relevance and fit of the standard curve to ensure the accuracy of quantitative analysis.

[0104] In summary, using geometric progression to set up the standard sample group and ensuring that there are at least 5 standard samples with different concentrations are important strategies to ensure the accuracy and reliability of the standard curve in ELISA experiments. These strategies help improve the accuracy of the quantitative analysis of the Klebsiella pneumoniae content in the sample.

[0105] In addition, the present application also provides a detection product in an embodiment, which adopts the method for determining the content of Klebsiella pneumoniae as described above;

[0106] The detection product includes at least one of a detection kit, a detection reagent and a detection device.

[0107] Example 1

[0108] In this embodiment, the content of Klebsiella pneumoniae is determined for the target sample.

[0109] Experimental methods:

[0110] (1) Set up different experimental groups, including a blank group, a test sample group, and a standard group.

[0111] The standard groups were set as: 10ng / mL, 20ng / mL, 30ng / mL, 40ng / mL, 50ng / mL, 60ng / mL, 70ng / mL, and 80ng / mL of glucose-6-phosphate dehydrogenase solution.

[0112] (2) For different experimental groups, the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase-labeled antibody and reaction substrate are added and incubated respectively to obtain incubated samples of each experimental group; the specific preparation method is shown in Table 1.

[0113] Table 1 Preparation method of each experimental group

[0114]

[0115] (3) Testing the incubated samples in each experimental group to obtain absorbance values.

[0116] The detection method adopted is ultraviolet spectrophotometry, and the detection wavelength is 450nm.

[0117] (4) Based on the absorbance value, a standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value is drawn.

[0118] In the standard group, the standard curve is obtained by plotting the concentration of glucose-6-phosphate dehydrogenase in each sample as the abscissa and the absorbance value of each sample as the ordinate.

[0119] Table 2 Absorbance values ​​of standard group

[0120]

[0121] The standard curve obtained by drawing the data in Table 2 is: y = 0.01x-0.0371, R 2 =0.992.

[0122] Here, x is the concentration of glucose-6-phosphate dehydrogenase (ng / mL), and y is the absorbance value at 450 nm.

[0123] (5) Substituting the absorbance value of the target sample into the standard curve to calculate the content of Klebsiella pneumoniae in the target sample.

[0124] Table 3 Test results of sample groups

[0125] NO. OD450 G6PD content in KP (ng / mL) KP content (μg) Sample 1 0.311 28.61 52.02 Sample 2 0.515 49.01 89.11 Sample 3 0.700 67.51 122.75

[0126] Note: Calculated based on 100μg KP containing approximately 2.75ng of G6PD.

[0127] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for determining the content of Klebsiella pneumoniae, characterized in that: The method for determining the content of Klebsiella pneumoniae is a method for determining the content of Klebsiella pneumoniae in a target sample by using glucose-6-phosphate dehydrogenase as a specific recognition site.

2. The method for determining the content of Klebsiella pneumoniae according to claim 1, characterized in that: include: Set up different experimental groups; For different experimental groups, the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase-labeled antibody and reaction substrate are added, and incubated respectively to obtain incubated samples of each experimental group; Detecting the incubated samples in each experimental group to obtain absorbance values; A standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value is drawn, and the Klebsiella pneumoniae content in the target sample is calculated according to the standard curve.

3. The method for determining the content of Klebsiella pneumoniae as claimed in claim 2, characterized in that: The different experimental groups are respectively: Blank group, test sample group and standard group.

4. The method for determining the content of Klebsiella pneumoniae as claimed in claim 3, characterized in that: The blank group contains a sterile solution; The sample group to be tested contains a suspension of the target sample; The standard set includes a glucose-6-phosphate dehydrogenase solution with a known concentration.

5. The method for determining the content of Klebsiella pneumoniae as claimed in claim 4, characterized in that: The method of drawing a standard curve of glucose-6-phosphate dehydrogenase concentration and absorbance value according to the detection data, and calculating the Klebsiella pneumoniae content in the target sample according to the standard curve, comprises: Using each sample in the standard group, the standard curve is drawn with the concentration of glucose-6-phosphate dehydrogenase as the abscissa and the absorbance value as the ordinate; The absorbance value of the target sample is brought into the standard curve to calculate the content of Klebsiella pneumoniae in the target sample.

6. The method for determining the content of Klebsiella pneumoniae as claimed in claim 3, characterized in that: The incubated samples of each experimental group are tested to obtain the absorbance value, and the detection method used includes at least one of ultraviolet spectrophotometry and enzyme labeling method; Preferably, the detection wavelength is 450 nm.

7. The method for determining the content of Klebsiella pneumoniae according to claim 3, characterized in that: For different experimental groups, the target sample, glucose-6-phosphate dehydrogenase solution, horseradish peroxidase labeled antibody and reaction substrate are added and incubated respectively, including: A sterile solution is added to the blank group; a glucose-6-phosphate dehydrogenase solution is added to the standard group; and a suspension of the target sample is added to the test sample group; Performing a first incubation on the blank group, the standard group and the test sample group; After adding horseradish peroxidase-labeled antibody to the blank group, the standard group and the test sample group, a second incubation is performed; Adding the reaction substrate to the blank group, the standard group and the sample group to be tested respectively for a third incubation, thus obtaining incubated samples of each experimental group; Preferably, the reaction substrate is 3,3',5,5'-tetramethylbenzidine; Preferably, the incubation temperatures of the first incubation, the second incubation and the third incubation are all: 37°C; Preferably, the incubation time of the first incubation is 20 minutes; Preferably, the incubation time of the second incubation is 30 minutes; Preferably, the incubation time of the third incubation is 20 minutes.

8. The method for determining the content of Klebsiella pneumoniae according to claim 2, characterized in that: The dilution ratio of the horseradish peroxidase-labeled antibody is 1:30,000.

9. The method for determining the content of Klebsiella pneumoniae as claimed in claim 3, characterized in that: In the standard group, the samples of glucose-6-phosphate dehydrogenase solution with different known concentrations have concentration values ​​that satisfy the orderly increasing arrangement of geometric progression from small to large; Preferably, the number of samples containing glucose-6-phosphate dehydrogenase solutions with different concentrations in the standard sample set is at least 5.

10. A detection product, characterized in that: The method for determining the content of Klebsiella pneumoniae according to any one of claims 1 to 9 is used; The detection product includes at least one of a detection kit, a detection reagent and a detection device.

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