Application of SLS in preparation of product capable of improving LAMP detection performance

Abstract

The invention relates to application of SLS in preparation of a product capable of improving LAMP detection performance, and belongs to the technical field of biology. The invention provides an application of sodium lauroyl sarcosinate (SLS) in preparation of a product capable of improving LAMP (loop-mediated isothermal amplification) detection performance. On one hand, the SLS has a target specificity optimization effect on detection of targets such as the monkey pox virus in an LAMP reaction system, and by adjusting the ion environment of the LAMP reaction system, the SLS can improve the specific amplification efficiency of the monkey pox virus target by 20% or above while completely inhibiting non-specific amplification; on the other hand, the SLS has the effect of improving the sensitivity and the detection rate on detection of targets such as the monkey pox virus in an LAMP reaction system, and under the low template concentration, the SLS can improve the detection rate of the monkey pox virus targets from 60% to 100% by inhibiting formation of primer dimers. Therefore, the SLS has a promising application prospect in LAMP-based nucleic acid detection.

Description

TECHNICAL FIELD

[0001] The present application relates to the application of SLS in the preparation of products capable of improving the detection performance of LAMP, belonging to the field of biotechnology. BACKGROUND

[0002] LAMP (Loop-mediated isothermal amplification) is a method that can achieve nucleic acid amplification at a constant temperature. This method does not require the thermal cycling steps of traditional PCR (Polymerase Chain Reaction), and compared with other nucleic acid amplification techniques, this method has the advantages of rapidity, high efficiency, strong specificity, easy judgment of results, no need for special instruments and independence from temperature changes.

[0003] However, the isothermal amplification reaction process of LAMP often has the problem of non-specific amplification (NS), which leads to false positive results, affecting the accuracy and reliability of LAMP detection. At present, the non-specific amplification is usually reduced by optimizing primer design, adjusting Mg 2+ concentration, adjusting reaction conditions, etc. However, these methods often have limited effect, and may affect the amplification efficiency while increasing the complexity of the experiment. Therefore, it is urgent to find a method that can effectively improve the specificity of LAMP without affecting its amplification efficiency and experimental complexity.

[0004] Sodium Lauryl Sarcosinate (SLS) is an anionic surfactant. The hydrophobic alkyl chain and hydrophilic sodium carboxylate in its molecular structure endow it with unique physicochemical properties, including the ability to reduce solution surface tension, dissolve lipid membranes, and regulate ionic environment. Currently, SLS has been used in PCR reactions for amplification optimization of complex templates (for DNA templates with high GC content or rich secondary structure, SLS can reduce the folding or aggregation of templates by disrupting the hydrophobic interactions between DNA strands, thereby improving the binding efficiency of primers to templates) and protection of polymerase activity (SLS can stabilize the ionic strength and buffer pH of the reaction system, reduce the irreversible damage of Taq enzyme or Bst DNA polymerase caused by high-temperature denaturation steps, prolong the thermal stability of the enzyme, and thus improve the success rate of long-fragment amplification). However, at present, the application of SLS in improving the specificity of LAMP has not been reported. SUMMARY

[0005] To solve the above problems, the present application provides the application of sodium lauryl sarcosinate in the preparation of products capable of improving the detection performance of LAMP, which includes improving the specific amplification efficiency, sensitivity and / or detection rate of LAMP detection.

[0006] In an embodiment of the present application, the product comprises an additive.

[0007] In an embodiment of the present application, the detection object of the LAMP detection comprises a virus; the virus is a DNA virus; the DNA virus comprises a monkeypox virus, a norovirus and / or a sapovirus.

[0008] The present application also provides a product capable of improving the performance of LAMP detection, wherein the components of the product comprise sodium lauroyl sarcosinate; and the improvement of the performance of LAMP detection comprises improving the specific amplification efficiency, sensitivity and / or detection rate of the LAMP detection.

[0009] In an embodiment of the present application, the product comprises an additive.

[0010] In an embodiment of the present application, the detection object of the LAMP detection comprises a virus; the virus is a DNA virus; the DNA virus comprises a monkeypox virus, a norovirus and / or a sapovirus.

[0011] The present application also provides a LAMP reagent, wherein the components of the LAMP reagent comprise sodium lauroyl sarcosinate, an isothermal amplification buffer and an enzyme component.

[0012] In an embodiment of the present application, the pH of the isothermal amplification buffer is 8.0-9.0.

[0013] In an embodiment of the present application, the components of the isothermal amplification buffer comprise Tris-HCl, (NH4)2SO4, KCl, MgSO4, dNTPs and a fluorescent dye.

[0014] In an embodiment of the present application, the fluorescent dye comprises at least one of SYTO-9 fluorescent dye, SYTO-11 fluorescent dye, SYTO-12 fluorescent dye, SYTO-14 fluorescent dye or SYTO-16 fluorescent dye.

[0015] In an embodiment of the present application, the concentration of the Tris-HCl in the LAMP reagent is 10-50 mM; the concentration of the (NH4)2SO4 in the LAMP reagent is 10-50 mM; the concentration of the KCl in the LAMP reagent is 50-150 mM; the mass percentage concentration of the dNTPs in the LAMP reagent is 0.8-1.6 mM; and the concentration of the fluorescent dye in the LAMP reagent is 0.02-2 μM.

[0016] In an embodiment of the present application, the components of the enzyme component comprise a DNA polymerase.

[0017] In an embodiment of the present application, the concentration of the DNA polymerase in the LAMP reagent is 8-60 U; and the concentration of the reverse transcriptase in the LAMP reagent is 40-80 U.

[0018] The present application also provides a LAMP detection kit, the components of which comprise the above-mentioned LAMP reagent and a LAMP primer set targeting the target nucleic acid.

[0019] In an embodiment of the present application, the detection object of the LAMP detection kit comprises a virus; the virus is a DNA virus; and the DNA virus comprises monkeypox virus, norovirus and / or sapovirus.

[0020] The present application also provides a LAMP-based nucleic acid detection method, which comprises using the above-mentioned LAMP detection kit to detect a sample to be tested.

[0021] In an embodiment of the present application, the nucleic acid detection method comprises mixing the LAMP reagent, the LAMP primer set targeting the target nucleic acid and the sample to be tested to obtain a LAMP reaction system; performing an amplification reaction on the LAMP reaction system; and after the amplification reaction is completed, determining whether the target nucleic acid exists in the sample to be tested according to the amplification curve obtained by the reaction (if the fluorescence signal exceeds the set threshold value and appears exponential growth, then the target nucleic acid exists in the sample to be tested).

[0022] In an embodiment of the present application, the concentration of the sodium lauroyl sarcosinate in the LAMP reaction system is 0.25-0.75 mM.

[0023] In an embodiment of the present application, the temperature of the incubation is 60-65℃, and the time is 30-60 min.

[0024] In an embodiment of the present application, the detection object of the nucleic acid detection method comprises a virus; the virus is a DNA virus; and the DNA virus comprises monkeypox virus, norovirus and / or sapovirus.

[0025] The present application also provides the use of sodium lauroyl sarcosinate or the above-mentioned product or the above-mentioned LAMP reagent or the above-mentioned LAMP detection kit in nucleic acid detection, which is not for the purpose of diagnosis and treatment of diseases.

[0026] In an embodiment of the present application, the detection object of the nucleic acid detection comprises a virus; the virus is a DNA virus; and the DNA virus comprises monkeypox virus, norovirus and / or sapovirus.

[0027] The technical scheme of the present application has the following advantages:

[0028] The application provides application of sodium lauroyl sarcosinate (SLS) in preparation of a product capable of improving LAMP detection performance, and the improvement of LAMP detection performance includes improving specific amplification efficiency, sensitivity and / or detection rate of LAMP detection. Traditional LAMP relies on additives such as betaine or DMSO to improve amplification efficiency, but these reagents have limited optimization effect on some targets (such as monkeypox virus), and high concentration is easy to inhibit the activity of Bst polymerase. In addition, LAMP is easy to produce non-specific amplification in low concentration template or complex sample, which leads to high false positive rate, delayed peak time and other problems. However, through research, it is found that on the one hand, SLS has target-specific optimization effect on the detection of targets such as monkeypox virus in the LAMP reaction system, and by adjusting the ionic environment of the LAMP reaction system, SLS (working concentration 0.5 mM) can significantly improve the specific amplification efficiency of monkeypox virus target by more than 20% while completely inhibiting non-specific amplification; on the other hand, SLS has sensitivity and detection rate improvement effect on the detection of targets such as monkeypox virus in the LAMP reaction system, and under low template concentration (6.25 Copies / Reaction), SLS (working concentration 0.5 mM) can improve the detection rate of monkeypox virus target from 60% to 100% by inhibiting primer dimer formation, breaking through the performance bottleneck of traditional additives, and combining electrophoresis and melting curve analysis, it is confirmed that the Tm value of the SLS optimization system is normal, and the non-template control group (NTC) has no amplification signal, which is significantly better than the existing LAMP method. Therefore, as an additive, SLS plays a dual function in LAMP technology, meets the rapid detection demand, and has great application prospect in nucleic acid detection based on LAMP. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 : Effect of SLS on LAMP amplification kinetics. Figure 1 , A: amplification curves of the positive group of Comparative Example 2 and Example 3; a: melting curves of the positive group of Comparative Example 2 and Example 3.

[0030] Figure 2 : Effect of SLS on LAMP amplification kinetics. Figure 2 , B: amplification curves of the positive group of Comparative Example 2 and Example 2; b: melting curves of the positive group of Comparative Example 2 and Example 2.

[0031] Figure 3 : Effect of SLS on LAMP amplification kinetics. Figure 3 , C: amplification curves of the positive group of Comparative Example 2 and Example 4; c: melting curves of the positive group of Comparative Example 2 and Example 4.

[0032] Figure 4 : Effect of SLS on LAMP amplification kinetics (significance analysis).

[0033] Figure 5 Effect of SLS on the detection rate of monkeypox virus. Figure 5 In the middle, A: amplification curve of the positive group of Comparative Example 2; a: dissolution curve of the positive group of Comparative Example 2; B: amplification curve of the positive group of Example 2; b: dissolution curve of the positive group of Example 2.

[0034] Figure 6 Effect of SLS on the detection rate of monkeypox virus (capillary electrophoresis spectrum).

[0035] Figure 7 Effect of SLS on the detection rate of monkeypox virus (significance analysis).

[0036] Figure 8 Effect of SLS on NS. Figure 8 In the middle, A: amplification curve of the positive group of Comparative Example 2; a: dissolution curve of the positive group of Comparative Example 2; B: amplification curve of the positive group of Example 2; b: dissolution curve of the positive group of Example 2.

[0037] Figure 9 Effect of SLS on NS (capillary electrophoresis spectrum). DETAILED DESCRIPTION

[0038] The following examples are provided to better enable those skilled in the art to which the application pertains to further understand the application and are not intended to limit the scope of the application or the meaning of the claims hereinafter. Any product falling within the scope of the present application is intended to be covered by the claims hereinafter, whether or not such product is expressly described in the examples or elsewhere herein.

[0039] In the following examples, the specific experimental procedures or conditions are not specified, which can be performed according to the conventional experimental procedures described in the literature in the art. The reagents or instruments used are not specified by the manufacturer, which are conventional reagent products that can be obtained by purchase.

[0040] In the following examples, the Bst DNA polymerase and dNTPs are purchased from Yixing Biotech (Shanghai) Co., Ltd., SYTO-16 fluorescent dye is purchased from Thermo Fisher Scientific, and MgSO4 is purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0041] Example 1: A LAMP detection kit for detecting monkeypox virus

[0042] The embodiment provides a LAMP detection kit for detecting monkeypox virus (MPXV), which is composed of 2.5x isothermal amplification buffer, 10x primers targeting MPXV, Bst DNA polymerase and sodium lauroyl sarcosinate (SLS);

[0043] The 2.5x isothermal amplification buffer is composed of 50mM Tris-HCl, 25mM(NH4)2SO4, 125mM KCl, 20mM MgSO4, 1.2mM dNTPs and 1μM SYTO-16 fluorescent dye (the solvent is nuclease-free pure water, and the pH is 8.8 at 25 DEG C).

[0044] The 10x primers targeting MPXV are composed of FIP primers with the nucleotide sequence shown in SEQ ID NO. 1, BIP primers with the nucleotide sequence shown in SEQ ID NO. 2, Loop F primers with the nucleotide sequence shown in SEQ ID NO. 3, F3 primers with the nucleotide sequence shown in SEQ ID NO. 4 and B3 primers with the nucleotide sequence shown in SEQ ID NO. 5 (see Table 1 for a specific formula).

[0045] Table 1 10x primer formula

[0046]

[0047] Embodiment 2: A nucleic acid detection method for detecting monkeypox virus

[0048] The embodiment provides a nucleic acid detection method for detecting monkeypox virus (MPXV), which comprises: preparing a LAMP reaction system (the concentration of sodium lauroyl sarcosinate in the LAMP reaction system is 0.5mM) according to the formula in Table 2 by using the LAMP detection kit in embodiment 1-1; after mixing and instantaneously centrifuging the LAMP reaction system, the LAMP reaction system is placed into a Thermofisher ABI QuantStudio1 real-time fluorescent quantitative PCR instrument to run an amplification reaction (see Table 3 for a LAMP reaction program); after the amplification reaction is completed, whether the monkeypox virus exists in the sample to be detected is determined according to the amplification curve obtained by the reaction.

[0049] Table 2 LAMP reaction system

[0050]

[0051]

[0052] In Table 2, the solvent of the 25mM SLS stock solution is nuclease-free water, 7.33mg SLS powder is weighed and dissolved in 1mL nuclease-free water to prepare a 25mM SLS stock solution, which is ready for use after 0.22μM filtration.

[0053] Table 3 LAMP reaction procedure

[0054]

[0055] Examples 3-4: A nucleic acid detection method for detecting monkeypox virus

[0056] The present example provides a nucleic acid detection method for detecting monkeypox virus (MPXV), which is based on Example 2, and the concentration of sodium laurylsarcosinate (SLS) in the LAMP reaction system is replaced by 0.25mM and 0.75mM respectively instead of 0.5mM.

[0057] Comparative Example 1: A LAMP detection kit for detecting monkeypox virus

[0058] The present comparative example provides a LAMP detection kit for detecting monkeypox virus (MPXV), which is based on Example 1-1, and sodium laurylsarcosinate (SLS) is removed.

[0059] Comparative Example 2: A nucleic acid detection method for detecting monkeypox virus

[0060] The present comparative example provides a nucleic acid detection method for detecting monkeypox virus (MPXV), which is based on Example 2, and the LAMP detection kit of Comparative Example 1 is used, and the concentration of sodium laurylsarcosinate (SLS) in the LAMP reaction system is 0mM.

[0061] Experimental Example 1: Effect of sodium laurylsarcosinate on LAMP isothermal amplification

[0062] Target: Monkeypox virus (MPXV)

[0063] Experiment 1: Effect of sodium laurylsarcosinate on amplification performance (25 Copies / Reaction)

[0064] The experimental process is as follows:

[0065] The monkeypox virus template used in this experiment was purchased from the National Standard Reference Material Research Center, with an original concentration of (2.74±0.46)×10^3 copies / μL, which was diluted with nuclease-free water to 5 copies / μL for standby, and the sample to be tested was obtained.

[0066] The nucleic acid detection method of Example 2 to Example 4 and Comparative Example 2 was used to detect the nucleic acid of the sample to be detected, and after the detection, the fluorescence signal results obtained by the real-time fluorescence quantitative PCR instrument were analyzed.

[0067] The experimental results are shown in Table 4 and Figures 1-4 The experimental results of this time added the melting curve, introduced the product Tm value to assist in judging specific amplification and non-specific amplification, and combined capillary electrophoresis to assist in reading under the condition that the Tm value could not be accurately judged.

[0068] Table 4 is the data of the fluorescence quantitative PCR instrument, and the CT value is the amplification time / cycle number when the fluorescence intensity reaches the set threshold. Under the same template concentration condition, the smaller the CT value, the higher the amplification efficiency, the better the specificity of the primer and the template, and the suitable reaction conditions (such as temperature, buffer composition, etc.), so that the Bst enzyme can more efficiently copy the target DNA sequence, thereby accumulating enough fluorescence signal in less amplification time, thereby reducing the CT value. Undetermined means not detected, that is, the fluorescence signal does not reach the threshold until the end of the reaction. In this experiment, Comparative Example 2 is used as the control group, and Examples 2 to 4 are used as the experimental group. The T / C ratio represents the ratio relationship between the CT value of the experimental group and the CT value of the control group. T / C<1 indicates that the amplification efficiency of the experimental group is better than that of the control group, and T / C>1 indicates that the amplification efficiency of the experimental group is weaker than that of the control group. The introduction of T / C as an amplification efficiency judgment basis in the isothermal amplification reaction can eliminate the small errors caused by template solution dilution in batch experiments, and uniformize the experimental results.

[0069] Figures 1-3 A, B, and C are the curve graphs of the fluorescence signal change with the amplification time (i.e., the amplification curve). The abscissa is the cycle number, and the ordinate is ΔRn, which represents the fluorescence signal change amount, i.e., the fluorescence intensity minus the baseline fluorescence intensity, which reflects the amount of target DNA produced in the LAMP reaction. The greater the fluorescence signal change amount, the more target DNA is produced. Figure 4 The significant analysis results obtained from the results of 5 independent repeated experiments are shown in Table 5.

[0070] The experimental results are analyzed as follows:

[0071] In terms of amplification efficiency, the T / C value of all experimental groups was less than 1, indicating that the amplification efficiency was better than that of the control group (without SLS) when the SLS addition amount was in the range of 0.25-0.75 mM. Among them, the T / C value of Example 3 (0.25 mM SLS) and Example 2 (0.5 mM SLS) was the lowest, and the amplification efficiency was improved by about 25%, and the coefficient of variation (CV) of CT value was less than 5%, indicating that the experimental results had high repeatability. In contrast, although the amplification efficiency of Example 4 (0.75 mM SLS) was slightly higher than that of the control group, the improvement was limited, and the coefficient of variation was large, indicating that its stability was poor (Table 4, Figures 1-3 ).

[0072] In terms of specificity, the Tm value of all experimental groups was stable at 83.937-84.250°C, which was consistent with the melting temperature of the control group, which confirmed that the addition of SLS did not introduce non-specific amplification products, and the specificity of LAMP reaction was maintained (Table 4).

[0073] To further verify the reliability of the experimental results, the experiment was repeated 5 times and subjected to significance analysis. The results showed that the T / C value of 0.25 mM and 0.5 mM SLS experimental groups was always less than 1, indicating that the peak time was significantly earlier than that of the control group, and the amplification efficiency was higher. The batch coefficient of variation (CV) was less than 10%, indicating that the experimental data had good consistency and repeatability. Among them, the difference between the 0.25 mM SLS group and the control group was marked as *** (P < 0.001), and the difference between the 0.5 mM SLS group and the control group was marked as **** (P < 0.0001), indicating that the improvement effect had statistical significance Figure 4 ).

[0074] Based on the above amplification efficiency improvement range (23.00% ± 5%) and statistical significance, it was determined that 0.5 mM SLS was the optimal working concentration and had good stability.

[0075] Experiment II: Effect of lauroylsarcosine sodium on amplification performance (6.25 Copies / Reaction)

[0076] The experimental process was as follows:

[0077] The monkeypox virus template used in this experiment was purchased from the National Standard Material Research Center, and the original concentration was (2.74 ± 0.46) × 10^3 copies / μL, which was diluted with nuclease-free water to 1.25 copies / μL for standby, i.e. the sample to be tested.

[0078] In this experiment, the nucleic acid detection method of Example 2 and Comparative Example 2 was used to detect the sample to be tested, and after the detection was completed, the fluorescence signal results obtained by the real-time fluorescence quantitative PCR instrument were analyzed.

[0079] Table 5 is the data of the fluorescence quantitative PCR instrument. Figure 5 is a curve graph of the fluorescence signal with the change of the amplification time (i.e. amplification curve). Figure 6 is the capillary electrophoresis spectrum of the LAMP amplification product. Figure 7 is the significance analysis result obtained according to the results of 5 independent repeated experiments. In this experiment, Comparative Example 2 is used as the control group of the experiment, and Example 2 is used as the experimental group.

[0080] The experimental results are analyzed as follows:

[0081] At a low template concentration of 6.25 Copies / Reaction, when the working concentration of SLS is 0.5 mM, in terms of specificity, there are 8 samples in the 20 parallel experiments of the control group (Comparative Example 2) whose Tm values are inconsistent with specific amplification (Table 5, Figure 5 ). Further analysis by capillary electrophoresis ( Figure 6 ), Sample 1 (Tm 84.707) is specific amplification as a spectrum control, it is found that Sample 2 (Tm 87.469) and Sample 12 (Tm 70.740) spectra are all presented as mixed peaks, confirming as non-specific amplification. In contrast, the Tm values of all 20 parallel experiments of Example 2 (0.5 mM SLS) remain consistent, and the capillary electrophoresis results all show specific amplification.

[0082] Comparing the experimental results of Example 2 and Comparative Example 2, the detection rate of Example 2 is significantly improved by 40%, the detection sensitivity reaches 6.25 Copies / Reaction, at the same time, the T / C is significantly reduced, and the amplification efficiency is obviously improved, which is highly consistent with the conclusion of Experiment One (Table 5).

[0083] The significance analysis of 5 repeated experiments shows that the difference between the experimental group and the control group has statistical significance (P < 0.0001, marked as ****), and the batch coefficient of variation (CV) is less than 10%, which confirms that the experimental results have good repeatability and stability ( Figure 7 ).

[0084] The above results show that the addition of 0.5 mM SLS can significantly improve the detection rate of monkeypox virus detection by 32.5% ± 7.5%, the detection limit is reduced to 6.25 Copies / Reaction, and the detection performance is significantly improved.

[0085] Experiment Three: Effect of Sodium Lauroyl Sarcosinate on Amplification Performance (0 Copies / Reaction)

[0086] The experiment is a no-template negative group (No-Template Control, NTC for short), and nuclease-free water is used instead of the template solution in the LAMP system as the sample to be tested.

[0087] The nucleic acid detection method of Example 2-Example 3 and Comparative Example 2 was used to detect the nucleic acid of the sample to be tested, and after the detection was completed, the fluorescence signal results obtained by the real-time fluorescence quantitative PCR instrument were analyzed.

[0088] Table 6 is the data of the fluorescence quantitative PCR instrument. Figure 8 is a curve graph of the fluorescence signal with the change of the amplification time (i.e. the amplification curve). Figure 9 is the capillary electrophoresis spectrum of the LAMP amplification product. In this experiment, Comparative Example 2 is used as the control group, and Examples 2-3 are used as the experimental group.

[0089] The experimental results are analyzed as follows:

[0090] Under the condition of no template negative at 0 Copies / Reaction, the monkeypox virus target primer group produces non-specific amplification with double Tm values under the condition of no template addition, one of which is 84℃ (Table 6, Figure 8 , which is close to the Tm value of specific amplification (84℃) in Experiment One, and needs to be further identified by capillary electrophoresis ( Figure 9 ). Capillary electrophoresis analysis shows that the peak type of the amplification product of Comparative Example 2 is significantly different from that of specific amplification, confirming that it is non-specific amplification. In contrast, the capillary electrophoresis spectrum of Example 2 (0.5mM SLS) does not detect amplification products, proving that SLS can effectively inhibit non-specific amplification. Further observation shows that with the increase of the addition concentration of SLS, its inhibitory effect shows an obvious concentration-dependent enhancement trend.

[0091] After 5 repeated experiments, the results all show that the experimental group with added SLS can completely inhibit the non-specific amplification phenomenon in the detection of monkeypox virus, and the inhibitory effect is stable and reliable. This finding confirms that SLS has a significant effect on improving the specificity of the LAMP reaction system, providing reliable technical support for improving the detection accuracy.

[0092] Table 4 Effect of SLS on LAMP amplification kinetics

[0093]

[0094] Table 5 Effect of 25mM SLS on detection rate of monkeypox virus

[0095]

[0096]

[0097] Table 6 Effect of SLS on NS

[0098]

[0099]

[0100] It is apparent that the above-described embodiments are merely illustrative in nature and not restrictive. Various modifications or changes can be made by those skilled in the art on the basis of the above description. It is not necessary to exhaust all the embodiments. The obvious changes or variations derived therefrom are still within the scope of the present application.

Claims

1. The use of sodium lauroyl sarcosinate in the preparation of a product capable of improving LAMP detection performance, characterized in that: Improving the LAMP detection performance includes improving the specific amplification efficiency, sensitivity and / or detection rate of the LAMP detection.

2. A product capable of improving LAMP detection performance, characterized in that: The product comprises sodium lauroyl sarcosinate; and improving the LAMP detection performance includes improving the specific amplification efficiency, sensitivity and / or detection rate of the LAMP detection.

3. A LAMP reagent, characterized in that The components of the LAMP reagent include sodium lauroyl sarcosinate, an isothermal amplification buffer and an enzyme component.

4. The LAMP reagent according to claim 3, wherein The pH of the isothermal amplification buffer is 8.0-9.

0.

5. The LAMP reagent according to claim 3 or 4, wherein The components of the enzyme component include DNA polymerase.

6. The LAMP reagent according to claim 5, wherein The concentration of the DNA polymerase in the LAMP reagent is 8 to 60 U; the concentration of the reverse transcriptase in the LAMP reagent is 40 to 80 U.

7. A LAMP detection kit, characterized in that The components of the LAMP detection kit include the LAMP reagent according to any one of claims 3 to 6 and a LAMP primer set targeting a target nucleic acid.

8. A method for nucleic acid detection based on LAMP, characterized in that: The nucleic acid detection method comprises: using the LAMP detection kit according to claim 7 to detect the sample to be tested.

9. The nucleic acid detection method according to claim 8, wherein The nucleic acid detection method includes: mixing a LAMP reagent, a LAMP primer set targeting a target nucleic acid, and a sample to be tested to obtain a LAMP reaction system; performing an amplification reaction on the LAMP reaction system; after the amplification reaction is completed, judging whether the target nucleic acid is present in the sample to be tested based on the amplification curve obtained by the reaction.

10. Use of sodium lauroyl sarcosinate or the product according to claim 2 or the LAMP reagent according to any one of claims 3 to 7 or the LAMP detection kit according to claim 8 in nucleic acid detection, characterized in that: The application is not for the purpose of diagnosis and treatment of diseases.

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