Formula of sample eluent

Abstract

The invention provides a formula of a sample eluent. The eluent comprises a surfactant, peptone with a concentration of 9 to 12 g / L, normal saline with a concentration of 7.2 to 9.6 g / L, tween-80 with a concentration of 0.8 to 1.2 g / L, disodium hydrogen phosphate with a concentration of 2.75 to 2.97 g / L and potassium dihydrogen phosphate with a concentration of 1.12 to 1.64 g / L. The eluent can effectively improve the sample recovery rate, and at least 90% of target substances can be successfully recovered after the eluent is treated by specific steps. In the use process, a dry and pollution-free sampling brush head needs to be completely immersed in the prepared eluent, then oscillation is carried out under set conditions to promote a target substance to be dissolved in the liquid, and finally the eluent is collected for subsequent analysis. The method is simple and convenient to operate, high in efficiency and very suitable for extracting and detecting various biological samples.

Description

Technical Field

[0001] The invention relates to the field of eluent production, in particular to a formula of a sample eluent. Background Art

[0002] At present, the collection of HPV test samples is mainly done by doctors. After the sample is collected by the sampling brush, it will be immediately broken into the preservation solution. The sample will be eluted and preserved during the transportation of the preservation solution. After arriving at the laboratory, it can be tested without eluting again. Although this is convenient for the examinee, it greatly prolongs the time for the sample to reach the laboratory. With the change of temperature in the four seasons, there will be extreme low and high temperatures. The use of preservation solution for storage and transportation may cause the sample to degrade under extreme high temperature conditions, resulting in a false negative report, which is very fatal to medical testing.

[0003] Therefore, if as many samples as possible for monitoring can be obtained when the samples are eluted and preserved during transportation of the preservation solution, the preservation time can be extended, thereby achieving the purpose of efficiently recovering the eluted samples. Summary of the invention

[0004] In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide a sample eluent formula

[0005] To achieve the above purpose, the technical solution provided by the present invention is:

[0006] A sample eluent formulation, the eluent comprising the following components:

[0007] Surfactants;

[0008] Peptone 9-12 g / L;

[0009] Normal saline 7.2-9.6 g / L;

[0010] Tween-80 0.8~1.2g / L;

[0011] Disodium hydrogen phosphate 2.75~2.97g / L;

[0012] Potassium dihydrogen phosphate 1.12~1.64g / L.

[0013] Furthermore, at 25°C, the pH value of the eluent is 7.3±0.1.

[0014] Furthermore, the surfactant is selected from at least one of nonionic, anionic or cationic surfactants.

[0015] Furthermore, the peptone is preferably 10.5 g / L.

[0016] Furthermore, the physiological saline is preferably 9 g / L.

[0017] Furthermore, the Tween-80 is preferably 1.1 g / L.

[0018] Furthermore, the sodium hydrogen phosphate is preferably 2.9 g / L.

[0019] Furthermore, the potassium dihydrogen phosphate is preferably 1.27 g / L.

[0020] A method for eluting a sample using an eluent comprises the following steps:

[0021] A) Prepare a sampling brush head containing a dry and stored target substance, and ensure that the sample on the sampling brush head is completely dry and not contaminated;

[0022] B) immersing the prepared sampling brush head into the eluent to ensure that the brush head is completely covered by the eluent;

[0023] C) placing the container containing the eluent and the sampling brush head on an oscillator, setting the oscillation frequency to 200 rpm and the duration to 3 minutes, so that the target substance is separated from the sampling brush head and dissolved in the eluent;

[0024] D) After stopping the shaking, take out the sampling brush head and collect the eluate for subsequent analysis or detection; experimental verification shows that this method can recover more than 90% of the samples for detection.

[0025] Compared with the prior art, the advantages of the present invention are: DNA samples adopt dry storage technology during storage, can be stored for a long time at room temperature, and do not need cold chain transportation, thus achieving long-term storage of samples. Before the DNA samples are tested in the laboratory, the formula of the present invention can obtain more than 90% of the samples for testing after 3 minutes of shaking and elution of the DNA samples, thus achieving the purpose of efficiently recovering the eluted samples from the dried DNA samples. DETAILED DESCRIPTION

[0026] Here, exemplary embodiments will be described in detail, and unless otherwise indicated, the embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. Instead, they are only examples consistent with some aspects of the present disclosure as detailed in the appended claims.

[0027] The specific implementation of the present invention is as follows:

[0028] A sample eluent formulation, the eluent comprising the following components:

[0029] A surfactant, selected from at least one of nonionic, anionic or cationic surfactants;

[0030] Peptone 9-12 g / L, preferably 10.5 g / L;

[0031] Normal saline 7.2-9.6 g / L, preferably 9 g / L;

[0032] Tween-80 0.8-1.2 g / L, preferably 1.1 g / L;

[0033] Disodium hydrogen phosphate 2.75-2.97 g / L, preferably 2.9 g / L;

[0034] Potassium dihydrogen phosphate 1.12-1.64 g / L, preferably 1.27 g / L.

[0035] At 25°C, the pH value of the eluent was 7.3±0.1.

[0036] According to the above formula, the present invention also provides a method for eluting a sample using an eluent, comprising the following steps:

[0037] A) Prepare a sampling brush head containing a dry and stored target substance, and ensure that the sample on the sampling brush head is completely dry and not contaminated;

[0038] B) Immerse the prepared sampling brush head in the eluent, making sure the brush head is completely covered by the eluent;

[0039] C) placing the container containing the eluent and the sampling brush head on an oscillator, setting the oscillation frequency to 200 rpm and the duration to 3 minutes, so that the target substance is separated from the sampling brush head and dissolved in the eluent;

[0040] D) After stopping the shaking, take out the sampling brush head and collect the eluate for subsequent analysis or detection; experimental verification shows that this method can recover more than 90% of the samples for detection.

[0041] In order to verify the above-mentioned coordination and operation steps, the present invention provides the following embodiments.

[0042] Embodiment 1:

[0043] Experimental design: Verify the efficiency of sample recovery by eluent

[0044] Purpose

[0045] The purpose of this experiment is to verify the efficiency of the above-mentioned eluent formula and its use method in recovering dry and stored target substances (such as simulated samples such as microorganisms and cell fragments), ensuring that more than 90% of the samples can be recovered for subsequent analysis or detection.

[0046] Experimental Materials

[0047] Elution solution: peptone 9g / L;

[0048] Normal saline 7.2 g / L;

[0049] Tween-80 0.8 g / L;

[0050] Disodium hydrogen phosphate 2.75 g / L;

[0051] Potassium dihydrogen phosphate 1.12g / L.

[0052] At 25°C, the pH of the eluent was 7.2.

[0053] Sampling brush head: Pre-smear evenly with target substance of known concentration and dry.

[0054] Oscillator: adjustable frequency up to 200 rpm.

[0055] Spectrophotometer or flow cytometer: used to quantitatively detect the concentration of target substances.

[0056] Sterile container: used to store eluent and samples.

[0057] Standard curve solution: used to establish the corresponding relationship between the concentration of the target substance and the detection signal.

[0058] Experimental procedures

[0059] Sample preparation:

[0060] Select one or more target substances and prepare them into a suspension of a certain concentration.

[0061] Using aseptic technique, spread the suspension evenly onto the sampling brush tip, ensuring even coverage.

[0062] Place the applied sampling brush head in a dry environment until it is completely dry.

[0063] Grouping and elution:

[0064] The dried sampling brush heads were randomly divided into several groups, with at least 3 replicates in each group.

[0065] Perform elution operations according to the above method B) to D), ensuring that each set of conditions is consistent.

[0066] Sample recovery and testing:

[0067] Collect the eluate after each set of elutions.

[0068] Measure the concentration of the target substance in the eluate using a spectrophotometer or flow cytometer.

[0069] At the same time, the concentration of the original suspension was determined as a control.

[0070] Data recording and analysis:

[0071] The average recovery rate of the target substance in each set of eluents was calculated, that is, the ratio of the concentration of the target substance in the eluent to the concentration of the original suspension multiplied by 100%.

[0072] Analyze the stability and repeatability of the recovery.

[0073] Experimental data recording and analysis

[0074] Table 1: Eluent recovery experimental data

[0075]

[0076] Experimental Results

[0077] According to the experimental data, the recovery rates of all groups were above 90%, with an average recovery rate of 91.83% and a standard deviation of 1.38%, indicating that the eluent formula and usage method have high stability and repeatability, and can effectively recover the dried and stored target substances.

[0078] Experimental analysis

[0079] The experimental results verified the effectiveness of the eluent formula and its usage method, and the recovery rate was stable and higher than 90%, which was in line with the experimental expectations.

[0080] Embodiment 2:

[0081] Experimental design: Same as Example 1

[0082] Experimental purpose: Same as Example 1

[0083] Experimental Materials

[0084] Novel eluent: surfactant selected from anionic surfactants;

[0085] Peptone 12 g / L;

[0086] Normal saline 9.6 g / L;

[0087] Tween-80 1.2 g / L;

[0088] Disodium hydrogen phosphate 2.97 g / L;

[0089] Potassium dihydrogen phosphate 1.64g / L.

[0090] At 25°C, the pH of the eluent was 7.4.

[0091] The rest is the same as in Example 1

[0092] Experimental procedures

[0093] Sample preparation:

[0094] Same as Example 1.

[0095] Grouping and elution:

[0096] Same as Example 1.

[0097] Sample recovery and testing:

[0098] Same as Example 1

[0099] Data recording and analysis:

[0100] Same as Example 1.

[0101] Experimental data recording and analysis

[0102] Table 2: Experimental data of eluent recovery rate in experimental group and control group

[0103]

[0104] Experimental Results

[0105] According to the experimental data, the recovery rate of the eluent used in the experimental group was above 94%, with an average recovery rate of 95%, while the recovery rate of the eluent used in the control group was between 84% and 86%, with an average recovery rate of 85%. The experimental results show that the eluent used in the experimental group of the present invention achieves the target in terms of recovery efficiency.

[0106] Experimental analysis

[0107] The significant difference between the experimental group and the control group shows that the new eluent designed in the present invention has obvious advantages in recovery efficiency.

[0108] Embodiment 3:

[0109] Experimental design: Same as Example 1

[0110] Experimental purpose: Same as Example 1

[0111] Experimental Materials

[0112] Surfactant, selected from cationic surfactant; peptone 10.5 g / L;

[0113] Normal saline 9g / L;

[0114] Tween-80 1.1 g / L;

[0115] Disodium hydrogen phosphate 2.9 g / L;

[0116] Potassium dihydrogen phosphate 1.27g / L.

[0117] At 25°C, the pH of the eluent was 7.3.

[0118] The rest is the same as in Example 1

[0119] Experimental procedures

[0120] Sample preparation:

[0121] Same as Example 1.

[0122] Grouping and elution:

[0123] Same as Example 1.

[0124] Sample recovery and testing:

[0125] Same as Example 1

[0126] Data recording and analysis:

[0127] Same as Example 1.

[0128] Experimental data recording and analysis

[0129]

[0130]

[0131] Experimental Results

[0132] According to the experimental data, the recovery rate of the experimental group using the eluent was above 91%, with an average recovery rate of 92%, while the recovery rate of the control group using the eluent was between 84% and 86%, with an average recovery rate of 85%. The experimental results met expectations.

[0133] Experimental analysis

[0134] The significant difference between the experimental group and the control group shows that the new eluent designed in the present invention has obvious advantages in recovery efficiency.

[0135] In summary, the formula of the present invention can obtain more than 90% of the sample for testing after 3 minutes of shaking and elution. It solves the problem of long-term storage of samples and the problem of long-distance storage under extreme temperature conditions, and efficiently recovers eluted samples, which has high social value.

[0136] The above is only one of the specific implementations of the present invention, but the technical features of the present invention are not limited thereto. Any modification, equivalent replacement or improvement made within the technical concept of the present invention shall be included in the protection scope of the present invention.

Claims

1. A sample eluent formulation, characterized in that: The eluent comprises the following components: Surfactants; Peptone 9-12 g / L; Normal saline 7.2-9.6 g / L; Tween-80 0.8~1.2g / L; Disodium hydrogen phosphate 2.75~2.97g / L; Potassium dihydrogen phosphate 1.12~1.64g / L.

2. The formulation according to claim 1, characterized in that At 25°C, the pH value of the eluent is 7.3±0.

1.

3. The formulation according to claim 1, characterized in that The surfactant is selected from at least one of nonionic, anionic or cationic surfactants.

4. The formulation according to claim 1, characterized in that: The peptone is preferably 10.5 g / L.

5. The formulation according to claim 1, characterized in that: The physiological saline solution is preferably 9 g / L.

6. The formulation according to claim 1, characterized in that: The Tween-80 is preferably 1.1 g / L.

7. The formulation according to claim 1, characterized in that: The sodium hydrogen phosphate is preferably 2.9 g / L.

8. The formulation according to claim 1, characterized in that: The potassium dihydrogen phosphate is preferably 1.27 g / L.

9. The method for eluting a sample using the eluent according to any one of claims 1 to 8, characterized in that: The steps include: A) Prepare a sampling brush head containing a dry and stored target substance, and ensure that the sample on the sampling brush head is completely dry and not contaminated; B) immersing the prepared sampling brush head into the eluent to ensure that the brush head is completely covered by the eluent; C) placing the container containing the eluent and the sampling brush head on an oscillator, setting the oscillation frequency to 200 rpm and the duration to 3 minutes, so that the target substance is separated from the sampling brush head and dissolved in the eluent; D) After stopping the shaking, take out the sampling brush head and collect the eluate for subsequent analysis or detection; experimental verification shows that this method can recover more than 90% of the samples for detection.

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