CN112763435A - Quantitative evaluation method for lethality of streptococcus pneumoniae - Google Patents
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- 241000193998 Streptococcus pneumoniae Species 0.000 title claims abstract description 58
- 229940031000 streptococcus pneumoniae Drugs 0.000 title claims abstract description 58
- 231100000225 lethality Toxicity 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 26
- 238000011158 quantitative evaluation Methods 0.000 title abstract description 12
- 238000010186 staining Methods 0.000 claims abstract description 25
- 241000894006 Bacteria Species 0.000 claims abstract description 23
- 230000001580 bacterial effect Effects 0.000 claims abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims abstract description 12
- 239000007791 liquid phase Substances 0.000 claims abstract description 11
- 238000002835 absorbance Methods 0.000 claims abstract description 8
- 239000011550 stock solution Substances 0.000 claims abstract description 8
- 238000007447 staining method Methods 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000012153 distilled water Substances 0.000 claims abstract description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 11
- 210000002966 serum Anatomy 0.000 claims description 10
- 239000002775 capsule Substances 0.000 claims description 9
- 241001052560 Thallis Species 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 238000009631 Broth culture Methods 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 230000003698 anagen phase Effects 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 238000004043 dyeing Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 claims description 3
- 229940107698 malachite green Drugs 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 5
- 238000005259 measurement Methods 0.000 abstract description 2
- 238000003556 assay Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 231100000518 lethal Toxicity 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 206010044008 tonsillitis Diseases 0.000 description 1
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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Abstract
The invention discloses a quantitative evaluation method for lethality of streptococcus pneumoniae, which comprises the following steps: staining by using 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50 Streptococcus pneumoniae stock solutions by adopting a liquid phase Streptococcus pneumoniae capsular staining method to prepare staining bacterial solutions with different concentrations; adjusting the zero value by using distilled water, carrying out color comparison on a plurality of groups of staining bacteria liquid with different concentrations in the step S1 respectively, measuring the absorbance at the position of 619nm wavelength, and drawing a standard curve; and (3) performing liquid-phase streptococcus pneumoniae capsular staining treatment on the sample to be detected, carrying out color comparison on the obtained staining bacteria liquid under the condition of 619nm wavelength, finding out the corresponding LD50 quantity on a standard curve according to the absorbance value obtained by color comparison, and obtaining the lethality measurement of the sample. The quantitative evaluation method for the lethality of the streptococcus pneumoniae provided by the invention does not need animal experiments, can complete determination within hours, and is a determination method which is simple, easy to operate and low in cost.
Description
Technical Field
The invention relates to the technical field of methods for determining the lethality of streptococcus pneumoniae, in particular to a quantitative evaluation method for the lethality of streptococcus pneumoniae.
Background
Streptococcus pneumoniae is a pathogenic gram-positive bacterium in human, and often causes respiratory tract infection in human, such as pneumonia, pharyngitis, tonsillitis and the like. The bacterium is a bacterium with a capsule, and the capsule is the most important lethal substance of the bacterium. The encapsulated streptococcus pneumoniae is called a virulent strain and the non-encapsulated streptococcus pneumoniae is called an avirulent strain. Thus, the capsule is closely and positively associated with its pathogenicity.
Half lethal dose (LD 50) is a commonly used measure for evaluating the lethality of bacteria. 1LD50 means: the minimum number of bacteria required to kill half of a given animal of a given weight or age by specifying the route of infection over a given period of time.
Since the past, the lethality of Streptococcus pneumoniae was evaluated, the LD50 assay was generally used. However, this assay requires at least tens of animals (usually mice) to be tested, is expensive, and requires several days to obtain the assay results.
In view of the above, it is an urgent need in the art to provide a simple, easy-to-operate, low-cost, and short-cycle measurement method.
Disclosure of Invention
The invention aims to provide a quantitative evaluation method for the lethality of streptococcus pneumoniae, which can complete determination within hours without animal experiments and is a determination method with simplicity, easy operation and low cost.
In order to solve the problems, the invention provides a quantitative evaluation method for the lethality of streptococcus pneumoniae, which comprises the following technical scheme:
a quantitative assessment method for lethality of streptococcus pneumoniae comprises the following steps:
step S1, staining by using a liquid-phase streptococcus pneumoniae capsular staining method with 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50 streptococcus pneumoniae bacterial stock solution to prepare staining bacterial solutions with different concentrations;
step S2, using distilled water for zero adjustment, carrying out color comparison on the multiple groups of staining bacteria liquid with different concentrations in the step S1 respectively, measuring the absorbance at the position of 619nm wavelength, and drawing a standard curve;
and step S3, performing liquid-phase streptococcus pneumoniae capsular staining treatment on the sample to be detected, carrying out color comparison on the obtained staining bacteria liquid under the condition of 619nm wavelength, finding out the corresponding LD50 number from the absorbance value obtained by color comparison on a standard curve, and obtaining the lethality measure of the sample.
Further, in steps S1 and S3, the liquid phase streptococcus pneumoniae capsule staining method includes the following steps:
centrifuging the streptococcus pneumoniae bacterial liquid for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated bacteria;
adding 10ml of 7% malachite green aqueous solution into the thallus precipitate, gently stirring, and dyeing at 37 ℃ for 20 minutes; centrifuging for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated thalli;
adding 10ml of normal saline into the thalli, centrifuging for 2 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated thalli;
adding 2ml of normal saline into the thallus precipitate, and gently and uniformly stirring to obtain the staining bacteria liquid.
Further, the concentration of pneumococcus at 1LD50 ═ 6.42 × 108CFU/mL, pneumococcus at 2LD50 concentration 1.28 × 109CFU/mL, pneumococcus at 3LD50 concentration 1.93 × 109CFU/mL, pneumococcus at a concentration of 4LD50 ═ 2.57X 109CFU/mL,5LD50 concentration of pneumococcus ═ 3.21X 109CFU/mL。
Further, in step S1, bacterial stock solutions with different concentrations were prepared as follows:
preparing a culture medium: 3 g of beef extract, 10 g of peptone and 5 g of sodium chloride, adding 1000ml of water, heating to dissolve, adjusting the pH to 7.5, and sterilizing for 20 minutes under the pressure of 103.4 kPa; adding sterile rabbit serum according to a concentration of 10% by aseptic operation to prepare a serum broth culture medium;
preparing a streptococcus pneumoniae culture solution: inoculating Streptococcus pneumoniae in logarithmic growth phase into serum broth, mixing, and culturing in 37 deg.C incubator for 24 hr;
preparing bacterial stock solutions with different concentrations: centrifuging the broth culture solution of streptococcus pneumoniae for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated bacteria; by dilution, 5 bacterial stocks of different concentrations were prepared, 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50, respectively.
Compared with the prior art, the quantitative evaluation method for the lethality of the streptococcus pneumoniae provided by the invention has the beneficial effects that:
according to the quantitative evaluation method for the lethality of the streptococcus pneumoniae, provided by the invention, capsular staining and spectrophotometric determination are used for quantitatively evaluating the lethality of the streptococcus pneumoniae, animal experiments are not needed, the detection can be completed within hours, and the method is a simple and easy-to-operate determination method with low cost.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a schematic diagram of a standard curve in the quantitative evaluation method for the lethality of Streptococcus pneumoniae provided by the present invention.
Detailed Description
In order to make the technical solutions in the embodiments of the present invention better understood and make the above objects, features, and advantages of the present invention more comprehensible, specific embodiments of the present invention are described below with reference to the accompanying drawings.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual values, and between the individual values may be combined with each other to yield one or more new ranges of values, which ranges of values should be considered as specifically disclosed herein.
The invention provides a quantitative evaluation method for lethality of streptococcus pneumoniae, which has the following experimental principle: the capsule of streptococcus pneumoniae is the most important lethal substance of the streptococcus pneumoniae, the capsular streptococcus pneumoniae is a virulent strain, and the non-capsular streptococcus pneumoniae is called an avirulent strain. The capsule is in close positive correlation with the lethality, so the size of the capsule is used as a quantitative index for evaluating the lethality of streptococcus pneumoniae.
First, experiment method
(I) culture Medium
The medium used in this example was serum broth which favours the production of the streptococcus pneumoniae capsule. The medium was prepared as follows:
3 g of beef extract, 10 g of peptone, 5 g of sodium chloride and 1000ml of water. Heating to dissolve, and adjusting pH to 7.5. Sterilization was carried out using 103.4kPa (15 lbs) for 20 minutes. Serum broth was prepared by adding sterile rabbit serum at 10% concentration in sterile procedures.
Culture of streptococcus pneumoniae
Inoculating streptococcus pneumoniae in logarithmic growth phase into a serum broth culture medium, uniformly mixing, and culturing in a 37-degree culture box for 24 hours to prepare the streptococcus pneumoniae culture solution.
(III) liquid-phase streptococcus pneumoniae capsular staining method
1. Centrifuging the broth culture solution of streptococcus pneumoniae for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated bacteria;
2. by dilution, 5 bacterial stocks of different concentrations were prepared, 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50, respectively. In this example, the concentration of pneumococcus at 1LD50 was 6.42 × 108CFU/mL, pneumococcus at 2LD50 concentration 1.28 × 109CFU/mL, pneumococcus at 3LD50 concentration 1.93 × 109CFU/mL, pneumococcus at a concentration of 4LD50 ═ 2.57X 109CFU/mL,5LD50 concentration of pneumococcus ═ 3.21X 109CFU/mL；
3. Adding 10ml of 7% malachite green aqueous solution into the thallus precipitate, gently stirring, and dyeing at 37 ℃ for 20 minutes; centrifuging for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated thalli;
4. adding 10ml of normal saline into the precipitate, centrifuging for 2 minutes at 5000 r/min without stirring, pouring out the supernatant, and leaving the precipitated thallus;
5. adding 2ml of normal saline into the thallus precipitate, and gently and uniformly stirring to obtain the staining bacteria liquid.
(IV) colorimetric determination
1. Staining by using 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50 bacteria stock solutions of streptococcus pneumoniae by adopting the liquid-phase streptococcus pneumoniae capsular staining method to prepare staining bacterial solutions with different concentrations;
2. adjusting the zero value by using distilled water, carrying out color comparison on 5 groups of staining bacteria liquid with different concentrations respectively, measuring the absorbance at the position of 619nm wavelength, and drawing a standard curve; in this example, a standard curve was plotted based on the total number of colonies of Streptococcus pneumoniae in 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50 as shown in FIG. 1;
3. and (3) performing liquid-phase streptococcus pneumoniae capsular staining treatment on the sample to be detected, carrying out color comparison on the obtained staining bacteria liquid under the condition of 619nm wavelength, finding out the corresponding LD50 quantity on a standard curve according to the absorbance value obtained by color comparison, and obtaining the lethality measurement of the sample.
According to the quantitative evaluation method for the lethality of the streptococcus pneumoniae, provided by the invention, capsular staining and spectrophotometric determination are used for quantitatively evaluating the lethality of the streptococcus pneumoniae, animal experiments are not needed, the detection can be completed within hours, and the method is a simple and easy-to-operate determination method with low cost.
The embodiments of the present invention are described in detail above with reference to the drawings, but the present invention is not limited to the described embodiments. Various changes, modifications, substitutions and alterations to these embodiments will occur to those skilled in the art without departing from the spirit and scope of the present invention.
Claims (4)
1. A quantitative assessment method for lethality of streptococcus pneumoniae is characterized by comprising the following steps:
step S1, staining by using a liquid-phase streptococcus pneumoniae capsular staining method with 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50 streptococcus pneumoniae bacterial stock solution to prepare staining bacterial solutions with different concentrations;
step S2, using distilled water for zero adjustment, carrying out color comparison on the multiple groups of staining bacteria liquid with different concentrations in the step S1 respectively, measuring the absorbance at the position of 619nm wavelength, and drawing a standard curve;
and step S3, performing liquid-phase streptococcus pneumoniae capsular staining treatment on the sample to be detected, carrying out color comparison on the obtained staining bacteria liquid under the condition of 619nm wavelength, finding out the corresponding LD50 number from the absorbance value obtained by color comparison on a standard curve, and obtaining the lethality measure of the sample.
2. The method for quantitatively evaluating lethality of streptococcus pneumoniae according to claim 1, wherein in steps S1 and S3, the liquid-phase streptococcus pneumoniae capsule staining method comprises the following steps:
centrifuging the streptococcus pneumoniae bacterial liquid for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated bacteria;
adding 10ml of 7% malachite green aqueous solution into the thallus precipitate, gently stirring, and dyeing at 37 ℃ for 20 minutes; centrifuging for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated thalli;
adding 10ml of normal saline into the thalli, centrifuging for 2 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated thalli;
adding 2ml of normal saline into the thallus precipitate, and gently and uniformly stirring to obtain the staining bacteria liquid.
3. The method for quantitatively evaluating lethality of streptococcus pneumoniae according to claim 1, wherein the concentration of pneumococcus at 1LD50 is 6.42 × 108CFU/mL, pneumococcus at 2LD50 concentration 1.28 × 109CFU/mL, pneumococcus at 3LD50 concentration 1.93 × 109CFU/mL, pneumococcus at a concentration of 4LD50 ═ 2.57X 109CFU/mL,5LD50 concentration of pneumococcus ═ 3.21X 109CFU/mL。
4. The method for quantitatively evaluating lethality of streptococcus pneumoniae according to any one of claims 1 to 3, wherein in step S1, bacterial stock solutions with different concentrations are prepared by:
preparing a culture medium: 3 g of beef extract, 10 g of peptone and 5 g of sodium chloride, adding 1000ml of water, heating to dissolve, adjusting the pH to 7.5, and sterilizing for 20 minutes under the pressure of 103.4 kPa; adding sterile rabbit serum according to a concentration of 10% by aseptic operation to prepare a serum broth culture medium;
preparing a streptococcus pneumoniae culture solution: inoculating Streptococcus pneumoniae in logarithmic growth phase into serum broth, mixing, and culturing in 37 deg.C incubator for 24 hr;
preparing bacterial stock solutions with different concentrations: centrifuging the broth culture solution of streptococcus pneumoniae for 8 minutes at the speed of 5000 r/min, pouring out supernatant, and leaving precipitated bacteria; by dilution, 5 bacterial stocks of different concentrations were prepared, 1LD50, 2LD50, 3LD50, 4LD50 and 5LD50, respectively.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5897993A (en) * | 1995-03-28 | 1999-04-27 | Idemitsu Kosan Company Limited | Method of determining the number of bacteria quickly and a device for determining the number of bacteria |
| CN104711315A (en) * | 2015-02-12 | 2015-06-17 | 北京华安科创生物技术有限公司 | Capsule staining method for streptococcus pneumoniae |
| US20170176325A1 (en) * | 2015-12-18 | 2017-06-22 | Abbott Laboratories | Spectral Differentiation of Histological Stains |
| CN108254238A (en) * | 2017-12-29 | 2018-07-06 | 乔治洛德方法研究和开发液化空气有限公司 | A kind of colouring method of filamentous microorganism and application thereof |
| CN110501336A (en) * | 2019-08-28 | 2019-11-26 | 北京智飞绿竹生物制药有限公司 | The capsular swelling colouring method of streptococcus pneumonia |
| CN112129610A (en) * | 2020-09-22 | 2020-12-25 | 无锡市第二人民医院 | Bacterial capsule staining method and application thereof |
-
2021
- 2021-04-01 CN CN202011183538.2A patent/CN112763435A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5897993A (en) * | 1995-03-28 | 1999-04-27 | Idemitsu Kosan Company Limited | Method of determining the number of bacteria quickly and a device for determining the number of bacteria |
| CN104711315A (en) * | 2015-02-12 | 2015-06-17 | 北京华安科创生物技术有限公司 | Capsule staining method for streptococcus pneumoniae |
| US20170176325A1 (en) * | 2015-12-18 | 2017-06-22 | Abbott Laboratories | Spectral Differentiation of Histological Stains |
| CN108254238A (en) * | 2017-12-29 | 2018-07-06 | 乔治洛德方法研究和开发液化空气有限公司 | A kind of colouring method of filamentous microorganism and application thereof |
| CN110501336A (en) * | 2019-08-28 | 2019-11-26 | 北京智飞绿竹生物制药有限公司 | The capsular swelling colouring method of streptococcus pneumonia |
| CN112129610A (en) * | 2020-09-22 | 2020-12-25 | 无锡市第二人民医院 | Bacterial capsule staining method and application thereof |
Non-Patent Citations (6)
| Title |
|---|
| 于威等: "《微生物学及检验技术》", 30 June 2008, 哈尔滨：黑龙江科学技术出版社, pages: 57 * |
| 周智兴等: "结核杆菌菌液浓度快速检测方法的建立", 《宜春学院学报》 * |
| 周智兴等: "结核杆菌菌液浓度快速检测方法的建立", 《宜春学院学报》, vol. 41, no. 09, 30 September 2019 (2019-09-30), pages 10 - 11 * |
| 周智兴等: "结核杆菌菌液浓度快速检测方法的建立", 宜春学院学报, vol. 41, no. 9, pages 10 - 11 * |
| 王欣等: "肺炎链球菌腹腔感染小鼠不同致死率动物模型的建立", 王欣等, vol. 47, no. 04, pages 120 - 121 * |
| 王燕等: "肺炎链球菌荚膜教学标本的制作方法研究", 中国当代医药, vol. 26, no. 6, pages 25 - 28 * |
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