Compositions and methods for detecting analytes

Abstract

1. An aqueous composition, e.g., a composition for use as a dissolution buffer, comprising zirconium oxide particles, a surfactant at a concentration of 0.005% (weight / volume) or greater, and a 10% to 10% (measured in deionized water at pH 8.45 and 20° C.) 4.2 and has a first affinity constant of 10 for magnesium 3.8 and a buffer. The aqueous composition has a pH of greater than or equal to 7.7 and less than 8.45, more particularly 7.8 to 8.3, in each case measured at 20° C. Methods of using the composition and kits comprising the components of the aqueous composition.

Description

[Background technology]

[0001] Patent Document 1 describes an aqueous lysis buffer composition for eliminating sample inhibition in an isothermal nucleic acid amplification reaction (i.e., a composition that contacts cells to lyse the cells and release their nucleic acids). This composition contains an organic iron chelating agent, ferric iron, a nonionic surfactant at a concentration of 0.005% (mass volume) or more, and 2-hydroxypropane-1,2,3-tricarboxylate, where the 2-hydroxypropane-1,2,3-tricarboxylate and the organic iron chelating agent are separate molecules. This aqueous composition has a pH of about 8.45 to 8.85.

[0002] Patent Document 2 discloses a method for amplifying nucleic acids involving the use of a lysis buffer. The lysis buffer may contain ferric ions and may further contain a reagent selected from the group consisting of a nanoparticle dispersion stabilizer, a non-ionic surfactant having a hydrophilic-lipophilic balance of about 11 to about 16, polyvinylpyrrolidone, magnesium sulfate heptahydrate, a fluorosurfactant, an indicator dye, and combinations of two or more of the above reagents. The lysis buffer is reported to have a pH of about 9.8 to 10.5 at 25°C.

[0003] Patent Document 3 discloses an aqueous composition for eliminating sample inhibition in a nucleic acid amplification reaction, which comprises a plurality of zirconium oxide particles, a nonionic surfactant at a concentration of 0.005% (mass / volume) or more, an organic iron chelating agent and a nanoparticle dispersion stabilizer, polyvinylpyrrolidone, or both. The composition has a pH of about 8.45 to 8.85.

[0004] Patent document 4 discloses reagents and methods for sterilizing reverse transcription reactions contaminated with nucleic acids produced from a previous reverse transcription / amplification reaction, obtained by mixing conventional and non-conventional nucleoside triphosphates. After sterilization, the nucleic acid to be amplified can be incubated in a liquid containing Tris-HCl (pH 8.3), KCl and EDTA, Tris-HCl (pH 8.3), KCl, DTT and MnCl2 and bicine KOAc and Mn(OAc)2 (pH 7.97). However, this liquid is not a lysis buffer, and the disclosure is not directed to reducing sample inhibition by matrix compounds. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] U.S. Pat. No. 10,604,787 [Patent Document 2] US Patent Application Publication No. 2019 / 0112637 [Patent Document 3] U.S. Pat. No. 10,619,189 [Patent Document 4] U.S. Pat. No. 5,693,517 Summary of the Invention

[0006] Disclosure Summary The above summary of the present disclosure is not intended to describe each disclosed embodiment or every implementation of the present disclosure. The following description illustrates exemplary embodiments in more detail. In several places in this application, guidance is provided through lists of examples, which examples can be used in various combinations. In each instance, the recited list serves only as a representative group and should not be interpreted as an exclusive list. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS In this application, terms such as "a," "an," and "the" are not intended to refer to a singular entity only, but include a general class of which specific examples may be used for illustration. The terms "a," "an," and "the" are used interchangeably with the phrases "at least one" and "one or more." The phrases "at least one of" and "including at least one of" following a list refer to any one of the items in the list and to a combination of two or more items in the list.

[0008] Terms such as "common," "generally," "often," "frequent," and "frequently" are used to refer to features typically employed in the present invention, but do not imply that the features so described were known or common prior to the present disclosure, unless specifically stated otherwise.

[0009] Unless otherwise stated, all pH values ​​in this disclosure and the appended claims refer to pH values ​​measured at 20°C.

[0010] Microorganisms or viruses, especially those found in food samples, can be detected by molecular methods. In such methods, for example those used by 3M Molecular Detection System (3M Company, St. Paul, MN, USA), the sample is contacted with an aqueous buffer solution, optionally after incubation. The buffer solution is a lysis buffer that lyses cells and releases nucleic acids from the cells. After contact with the lysis buffer, amplification of the nucleic acids can be performed.

[0011] Patent Documents 1, 3 and 2 disclose that zirconium oxide particles may be added to the lysis buffer to reduce or eliminate the effect of food matrices, i.e., compounds from food samples that may be present in the sample and interfere with the amplification or detection of nucleic acids. Tris, an amine-containing cationic buffer, is the preferred buffering agent in these disclosures, which indicates that the buffer must have a pH of 8.45 to 8.85, which is the buffer range for Tris buffers.

[0012] The present disclosure recognizes several problems with prior art dissolution buffers. First, some samples may not be sufficiently stable in the pH range of 8.45-8.85 disclosed in the aforementioned references, but may be more stable in a lower, i.e., closer to neutral, pH range. Second, carbon dioxide in the air may act as an acid (by becoming carbonic acid upon contact with or dissolution in an aqueous liquid), thus neutralizing the relatively high pH of the prior art buffers. This effect may be somewhat less at lower pHs. Third, anionic or cationic buffers such as Tris buffers are somewhat limited in the amount that can be used in aqueous liquids, as they may interfere with the nucleic acid amplification or detection process. Thus, prior art buffers have low buffering capacity. Low buffering capacity limits the types of food matrices that can be tested, since some food matrices are very acidic or basic, which cannot be converted to an acceptable pH for amplification by prior art buffers. Fourth, a fast time to result is advantageous, regardless of pH.

[0013] Briefly, the solution to these and other problems is an aqueous composition comprising zirconium oxide particles, a surfactant at a concentration of 0.005% (weight / volume) or greater, and 10% by weight of ferric iron. 4.2 and has a first affinity constant of 10 for magnesium 3.8and a buffer, optionally having a concentration of 40 mM or more, further optionally having a concentration of 40 mM to about 200 mM, and still further optionally having a concentration of 40 mM to 150 mM. The aqueous composition has a pH of 7.7 to 8.45, more particularly 7.8 to 8.3, in each case measured at 20° C. The aqueous composition is typically a dissolution buffer, such as a dissolution buffer composition.

[0014] The solution also resides in a nucleic acid amplification method comprising the steps of: a) contacting an aqueous composition described herein with a composition comprising a microorganism or virus to form a mixture; b) lysing the microorganism or virus in the mixture to form a lysis mixture; and c) subjecting at least a portion of the lysis mixture to a nucleic acid amplification process.

[0015] The solution also includes a kit comprising: a plurality of zirconium oxide particles; a non-ionic surfactant; and 10 4.2 and has a first affinity constant of 10 for magnesium 3.8 and a buffer having a buffer range ranging from pH 7.8 or less to pH 8.2 or more at 20° C. Each component in the kit can be provided as a dry component for the user to dissolve in water, e.g., to form an aqueous composition as described herein. Alternatively, one or more components of the kit can be dissolved in water in the kit, with the dry components, if any, added later.

[0016] aqueous composition The aqueous composition may be a solution or a dispersion. In the case of a dispersion, zirconium oxide particles are typically dispersed in the composition. The zirconium oxide particles are nanoparticles in some embodiments. In certain embodiments, the zirconium oxide particles have an average particle size of 500 nm or less, more particularly 250 nm or less, even more particularly 100 nm or less, respectively, as measured by photon correlation spectroscopy as described herein below. The zirconium oxide particles optionally have a surface area (in m2) of at least 10, for example 10 to 600, particularly 25 to 600, more particularly 50 to 600, even more particularly 100 to 600, still more particularly 200 to 600, even more particularly 300 to 600, most particularly 400 to 600. 2 / L).

[0017] In either case, the particle size can be measured by photon correlation spectroscopy (PCS) according to the method described in the "Test Methods" section of the specification of US Pat. No. 864,710. Specifically, a PCS device equipped with a red laser (wavelength 633 nm), such as the Zeta Sizer-Nano Series, Model ZEN 3600, can be used. The sample is placed in a 1 cm square cuvette to an appropriate liquid depth, such as 10 to 14 mm. The liquid depth depends on the dimensions of the device used. The cuvette is then placed in the device and equilibrated at 25°C. The parameters of the device can be set as follows: refractive index of dispersant: 1.3330, viscosity of dispersant: 0.8872 mPa·sec, refractive index of material: 2.10, and absorption value of material 0.10 units. The size measurement procedure of the device can then be performed according to the device's instruction manual. In most PCS instruments, the laser beam position and attenuator settings are automatically adjusted to obtain the best measurement of particle size, but if your instrument does not automatically adjust them, they can be optimized according to the instrument's instructions or standard instrument optimization techniques to obtain the best measurement of particle size (e.g., most reproducible measurements, best signal-to-noise ratio, etc.). In many cases, commercial zirconium oxide particles are available with a labeled particle size (e.g., measured by the manufacturer), and measurement of particle size is generally not necessary, since the manufacturer's indication of particle size (e.g., on the product label) can be trusted.

[0018] A stabilizer can be added to stabilize any of the aforementioned zirconium oxide particles. Most commonly, the stabilizer is citric acid or its salts, such as potassium citrate, ferric ammonium citrate, and the like. Other stabilizers can be used as long as they do not interfere with the amplification or detection of nucleic acids. In some cases, no stabilizer is required, since some zirconium oxide particles can form a stable dispersion at the required pH value without a stabilizer.

[0019] In any of the above cases, the pH (measured at 20° C.) may be greater than 7.7 and less than 8.45. In particular, the pH (measured at 20° C.) may be greater than 7.7, greater than 7.8, greater than 7.9 or greater than 8.0. In particular, the pH (measured at 20° C.) may be less than 8.45, less than 8.4, less than 8.3 or less than 8.2. Most commonly, and most particularly, the pH is between 7.8 and 8.3.

[0020] The buffer particularly comprises at least one zwitterionic compound, meaning that the compound is present in zwitterionic form at the pH of the composition. Without being bound by theory, the inventors hypothesize that cationic, anionic or non-ionic buffers may coordinate with nucleic acid strands and thus interfere with the amplification or detection of microorganisms in the sample. A particularly useful zwitterionic compound is bicine. Thus, the buffer is particularly bicine.

[0021] The buffer, particularly the zwitterionic buffer, most particularly bicine, can have any suitable concentration, but will usually have a concentration of 40 mM or more, more particularly 40 mM to 200 mM, even more particularly 40 mM to 150 mM, and even more particularly 50 mM to 150 mM. Buffers at these concentrations provide higher buffering capacity than that available with cationic or anionic buffers such as Tris, which at high concentrations interfere with nucleic acid amplification or detection processes.

[0022] The iron chelating agent is 10 for ferric iron. 4.2 and has a first affinity constant of 10 for magnesium 3.8The first and second affinity constants are measured in deionized water at pH 8.45 and 20°C. Thus, the iron chelating agent has a greater affinity for ferric iron than for magnesium. The iron chelating agent is typically an organic iron chelating agent, which means that the iron chelator is an organic compound, but this does not mean that the organic iron chelating compound chelates only organic iron compounds.

[0023] Any suitable organic iron chelating agent can be used. Most typically, the organic iron chelating agent comprises ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), N,N',N',N'-tetrakis(2-pyridinylmethyl)ethane-1,2-diamine, 1,2-bis(O-aminophenoxy)ethane-N,N,N'-tetraacetic acid, N-(2-hydroethoxyethyl)ethylenediamine-N,N',N'-triacetic acid, a salt of any of the foregoing, or a hydrate of any of the foregoing. Most commonly, a salt such as a sodium or potassium salt or a mixed sodium / potassium salt of the organic iron chelating agent is used, especially the potassium salt. EGTA is most commonly used, most especially the potassium salt of EGTA.

[0024] Optionally, in any of the embodiments mentioned herein, the composition may require ferric iron. When included, the ferric iron typically has a concentration of 50 to 385 micromolar, for example at least 110 micromolar, at least 165 micromolar, at least 220 micromolar, at least 275 micromolar or at least 330 micromolar; in any case, the maximum concentration may be 385 micromolar. When ferric iron is included, the molar ratio of ferric ion of the ferric iron to the organic iron chelating agent is typically 0.04 to 0.28, more particularly 0.14 to 0.18.

[0025] The at least one non-ionic surfactant can be any suitable non-ionic surfactant that provides a stable formulation, e.g., does not precipitate components intended to be dissolved and suspends components intended to be suspended, for a commercially acceptable time period after preparation of the composition. Particularly useful non-ionic surfactants include those having a hydrophilic-lipophilic balance (HLB) of 11 to 16. This HLB range promotes the activity of DNA polymerases used in nucleic acid amplification such as PCR and LAMP. Non-limiting examples of specific nonionic surfactants that can be used include those available under the TRITON trade name, such as TRITON X-100, TRITON X-114, TRITON X-405, TRITON X-101, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters (such as those available under the TWEEN trade name), polyoxyethylene alkyl esters (such as those available under the BRIJ trade name), nonylphenols, lauryl alcohol, polyethylene glycols, polyoxyethylene-polyoxypropylene block copolymers, polyoxyethylene alkylamines, polyoxyethylene fatty acid bisphenyl ethers, and fluorosurfactants (such as those sold under the NOVEC trade name by 3M Company St. Paul MN USA).

[0026] The non-ionic surfactant can be present at any suitable concentration, for example, a concentration that meets one or more of the above criteria, or other criteria required by the particular end use. Typically, a concentration of 0.005% (w / v) to 0.3% (w / v), for example 0.01% to 0.3% (w / v), is used.

[0027] Magnesium ions, potassium ions or both may also be used in the composition. These may facilitate downstream nucleic acid amplification of the sample, for example by PCR, e.g. qPCR, LAMP, etc. The amount of magnesium ions, if used, is typically 1 mM to 15 mM. The amount of potassium ions, if used, is typically 5 mM to 500 mM, e.g. 20 mM to 60 mM. In particular, magnesium ions are included as a component of magnesium salts, such as magnesium sulfate or its hydrates, more particularly magnesium sulfate heptahydrate.

[0028] Optionally, the composition may further comprise one or more additional components, if used, most commonly one or more of an indicator dye, a preservative, an enhancer for the LAMP reaction, an enhancer for the qPCR reaction, or a fluorosurfactant.

[0029] If an indicator dye is used, it may be any dye suitable for the application, for example a dye suitable for detecting one or more microorganisms of interest. Many indicator dyes are known in the art and in principle any may be used. A particularly common dye is cresol red. An indicator dye is not always necessary, as some detection systems do not rely on an indicator dye, and in some cases the desired dye may be added in a downstream processing step.

[0030] A variety of preservatives suitable for use in biological systems are known, and one of skill in the art can select which, if any, to use depending on the desired end use. One particularly useful preservative is methylisothiazolinone.

[0031] Enhancement agents for facilitating LAMP or qPCR reactions are also known in the art and can be selected depending on the desired end use, such as the type of nucleic acid amplification used, for example, sulfates such as magnesium sulfate or ammonium sulfate or their hydrates, potassium chloride, etc.

[0032] kit The compositions can be pre-prepared for the end user, or a kit can be provided to the end user to prepare the composition from the components of the kit and, optionally, water provided by the user. Thus, the kit can include a plurality of zirconium oxide particles, which can be any of the particles described above with reference to the compositions. The zirconium oxide particles can be provided as a solid to be dispersed in water, such as deionized water, or as an aqueous dispersion.

[0033] The kit contains 10 for ferric iron. 4.2 and has a first affinity constant of 10 for magnesium 3.8 The composition may further comprise an organic iron chelating agent having a second affinity constant less than 100%, where the first affinity constant and the second affinity constant are measured in deionized water at pH 8.45 and 20° C. Any of the organic iron chelating agents described above with reference to the composition may be used. EGTA is the most common. The organic iron chelating agent may be dispersed or provided in water, such as deionized water.

[0034] The kit can further include a buffer. The buffer most commonly has a buffering range that ranges from pH 7.7 or less to pH 8.2 or more at 20°C. Thus, the buffer can typically provide buffering capacity in the pH range of the composition, which range is discussed in detail above. The buffer is typically zwitterionic, most particularly bicine, as discussed in detail above. The buffer can be provided as a solid that is reconstituted with water, such as deionized water, or can be dissolved in water.

[0035] Similarly, the kit may provide at least one of a nanoparticle dispersion stabilizer and an indicator dye, a preservative, an enhancer for a LAMP reaction, an enhancer for a qPCR reaction, or a fluorosurfactant, and / or a ferric salt, any of which may be dispersed in water or may be provided as a solid that is subsequently dispersed in water.

[0036] Most commonly, the plurality of zirconium oxide particles, the non-ionic surfactant, and at least one of the organic iron chelating agent are disposed in an aqueous liquid having a pH (measured at 20° C.) greater than 7.7 and less than 8.45. In particular cases, the pH (measured at 20° C.) can be greater than 7.7, greater than 7.8, greater than 7.9, or greater than 8.0. In particular, the pH (measured at 20° C.) can be less than 8.45, less than 8.4, less than 8.3, or less than 8.2. Most commonly, and most particularly, the pH is between 7.8 and 8.3. The remaining components are most commonly provided as solids that are subsequently added to the aqueous liquid.

[0037] How to use The compositions and kits disclosed herein are most commonly used in nucleic acid amplification methods.The kits can be used by first combining the components of the kit with each other and / or with water, particularly deionized water or reverse osmosis purified water, to form the compositions disclosed herein.The compositions can then be used according to the general methods known in the art, such as those described in Patent Document 3.

[0038] Briefly, the composition described herein can be contacted with a sample that contains or is suspected to contain a microorganism or virus to form a mixture. The sample can be optionally incubated before this contacting step, especially when it is necessary to do so to increase the number of microorganisms or viruses. The microorganisms or viruses can then be lysed to form a lysis mixture. The lysis mixture can then be subjected to a nucleic acid amplification process to amplify one or more nucleic acids that were present in the microorganisms or viruses.

[0039] The dissolution step is typically thermal dissolution, which is most often accomplished by heating the mixture to 80-115° C. for 5-30 minutes.

[0040] The nucleic acid amplification method may be any known method, but PCR, such as qPCR or LAMP, is the most common. qPCR amplification is known and is described, for example, in the article "Real-time PCR in the microbiology laboratory" by Mackay, I., European Society of Clinical Microbiology and Infectious Diseases 2004 (190). LAMP amplification is described, for example, in U.S. Pat. No. 9,090,168. The results of amplification such as LAMP amplification can be detected by known methods such as those described in the article "Novel bioluminescent quantitative detection of nucleic acid amplification in real-time" by Gandelman, O. et al., PLoS One, Nov 30;5(11).

[0041] Working Example Zirconium oxide nanoparticle dispersion (5 wt % in water, average particle size ≦100 nm (BET), product number 643122) was obtained from Sigma Aldrich Company, St. Louis, MO.

[0042] Citric acid (product number C1909), polyvinylpyrrolidone (product number P5288), TRITON X-100 surfactant (product number T8787), bicine (product number B8660), potassium acetate (P1190), potassium hydroxide (product number 60370), EGTA (product number 03777), magnesium heptahydrate (product number 63138) and PROCLIN 950 (product number 46878-U) were all obtained from Sigma Aldrich Company.

[0043] The pH of the compositions was measured at 20° C. using an ACCUMET AE150 benchtop pH meter (obtained from Thermo Fisher Scientific, Waltham, Mass.) equipped with an ACCUMET gel-filled polymer body pH / ATC double junction combination electrode (mercury-free). Measurements were taken within 24 hours of sample preparation.

[0044] Example 1. A suspension composition was prepared by adding each of the components shown in Table 1 in the order specified to deionized water and mixing, the pH of this composition being 8.1.

[0045] [Table 1]

[0046] Comparative example A. A composition was prepared as described in Example 1, except that the following components were not included in the composition: citric acid, zirconium oxide dispersion, EGTA, magnesium sulfate heptahydrate. The pH of this composition was 8.3.

[0047] Example 2. A composition was prepared by adding each component to deionized water in the order specified in Table 2. The pH of this composition was 8.3. Compared to the composition of Example 1 (Table 1), this composition (Table 2) had higher concentrations of bicine and potassium hydride and lower concentrations of potassium acetate to increase buffering capacity.

[0048] [Table 2]

[0049] Comparative example B. A composition was prepared as described in Example 2, except that the following components were not included in the composition: citric acid, zirconium oxide dispersion, EGTA, magnesium sulfate heptahydrate. The pH of this composition was 8.3.

[0050] Comparative example C. A composition was prepared as described in Example 2 of Patent Document 3. The pH of this composition was 8.7.

[0051] Example 3. Loop-Mediated Isothermal Amplification (LAMP) - Bioluminescence Detection Assays Using the Compositions of Examples 1 and 2, and Comparative Examples A-C Raw ground chicken (32 g) and Buffered Peptone Water Rich Medium (BPW-ISO, 162 mL, preheated to 41.5° C., obtained from 3M Company, St. Paul, MN) were combined in a Nasco WHIRL-PAK homogenizer filter bag (product number 01318, obtained from Thermo Fisher Scientific) and mixed for 2 minutes at 230 rpm (revolutions per minute). The sample was incubated at 41.5° C. for 6 hours. Selected aliquots (580 microliters) of the compositions of Examples 1 and 2, and Comparative Examples A through C, were individually added to separate 1.1 mL AXYGEN minitubes (product number MTS-11-12-CR, obtained from Corning Inc., Corning, NY) (i.e., a single composition was added to each tube). Then, 20 microliters of concentrated sample from the homogenizer bag was added to each tube.

[0052] For nucleic amplification and bioluminescence detection, each tube was heated in a 100°C heat block for 15 minutes and cooled to approximately 40°C, after which a 20-microliter aliquot of the mixture was added to a reaction tube containing a universal matrix control pellet (product number MDMC96NA, available from 3M Company). Three reaction tubes (n=3) were prepared for each composition. After dissolving the pellets, each reaction tube was analyzed using a 3M Molecular Detection Instrument (product number MDS 100; available from 3M Company) and the bioluminescence signal was recorded according to the manufacturer's instructions. The maximum bioluminescence signal (relative light units (RLU)) and the reaction time at which the maximum bioluminescence signal occurred were recorded. The results are reported in Tables 3 to 5 as the average of three tests.

[0053] [Table 3]

[0054] [Table 4]

[0055] [Table 5]

[0056] Example 4. LAMP-bioluminescence detection assay using compositions at various pH values The pH of compositions prepared according to the procedure of Example 1 (Table 1) was adjusted with various amounts of glacial acetic acid to provide five separate compositions having a pH of either 7.4, 7.6, 7.8, 8.0 or 8.2. The reaction time at which the maximum bioluminescent signal occurred was determined according to the procedure of Example 3. The results are reported in Table 6 as the average of three tests.

[0057] [Table 6]

Claims

1. An aqueous composition, wherein the aqueous composition comprises: zirconium oxide particles; a surfactant having a concentration of 0.005% (mass / volume) or more; an organic iron chelating reagent having a first affinity constant of 104.2 or more with respect to ferric iron and a second affinity constant of less than 103.8 with respect to magnesium, wherein the first affinity constant and the second affinity constant are measured in deionized water at 20 °C and pH 8.45, the organic iron chelating reagent; a buffer comprising at least one zwitterionic compound and having a concentration of 40 mM or more and, the composition has a pH greater than 7.7 and less than 8.45, optionally 7.8 to 8.3, when measured at 20 °C, the aqueous composition.

2. The aqueous composition according to claim 1, wherein the buffer comprises at least one zwitterionic compound, the buffer has a concentration of 40 mM or more, further optionally has a concentration of 40 mM to 200 mM, and still further optionally has a concentration of 40 mM to 150 mM.

3. The aqueous composition according to claim 2, wherein the at least one zwitterionic compound comprises bicine.

4. The aqueous composition according to claim 1, further comprising citric acid or a salt thereof.

5. The aqueous composition according to claim 1, further comprising a magnesium salt, optionally wherein the magnesium salt is magnesium sulfate or a hydrate thereof, and further optionally magnesium sulfate heptahydrate.

6. The aqueous composition according to claim 1, further comprising ferric iron ions.

7. The aqueous composition according to claim 1, wherein the zirconium oxide particles each have an average particle size of 500 nm or less, optionally 250 nm or less, when measured by the photon correlation spectroscopy described herein.

8. The aqueous composition according to claim 1, further comprising at least one of an indicator dye, a preservative, an enhancer for LAMP reaction, an enhancer for qPCR reaction, or a fluorine-based surfactant.

9. A nucleic acid amplification method, comprising: a) contacting the composition according to any one of claims 1 to 8 with a sample containing a microorganism or a virus to form a mixture; b) lysing the microorganism or the virus in the mixture to form a lysed mixture; and c) subjecting at least a part of the lysed mixture to a nucleic acid amplification process The method.

10. The method according to claim 9, further comprising the step of incubating the composition containing the microorganism or virus in a growth medium before step a).

11. A kit comprising: A plurality of zirconium oxide particles; A non-ionic surfactant; An organic iron chelating reagent having a first affinity constant of 104.2 or more with respect to ferric iron and a second affinity constant of less than 103.8 with respect to magnesium, wherein the first affinity constant and the second affinity constant are measured in deionized water at pH 8.45 and 20 °C, the organic iron chelating reagent; A buffer, the buffer containing at least one zwitterionic compound and optionally having a concentration of 40 mM or more, the buffer having a buffering region ranging from pH 7.8 or less to pH 8.2 or more at 20 °C The kit comprising.

12. The kit according to claim 11, further comprising a nanoparticle dispersion stabilizer.

13. The kit according to claim 11, wherein at least one of the plurality of zirconium oxide particles, the non-ionic surfactant and the organic iron chelating reagent is disposed in an aqueous liquid having a pH greater than 7.7 and less than 8.45, optionally 7.8 to 8.3, when measured at 20 °C.

14. The kit according to claim 11, further comprising an indicator dye, a preservative, an enhancer for LAMP reaction, an enhancer for qPCR reaction or a fluorine-based surfactant.

15. The kit according to claim 11, wherein the kit further comprises a ferric iron salt.

16. The kit according to claim 11, wherein the at least one zwitterionic compound comprises bicine.