CN-122029290-A - Method and system for rapid detection of yeast

Abstract

A method of preparing a sample for an assay for rapid quantification of biological contaminants. The method includes adding a sample to a vessel and inserting the vessel into a centrifuge. The method further includes centrifuging the sample to produce a first supernatant and a first precipitate, discarding the first supernatant, and dissolving the first precipitate in a first volume of buffer to produce a first suspension volume. The method further includes centrifuging the first suspension volume to produce a second supernatant and a second precipitate, discarding the second supernatant, and dissolving the second precipitate in a second volume of buffer to produce a final sample suspension. The method further comprises adding the final sample suspension to an assay for rapid quantification of microbial contaminants.

Inventors

• S. Laws
• P. Safco
• A. Puerta Gomez

Assignees

• 可口可乐公司

Dates

Publication Date

20260512

Application Date

20240930

Priority Date

20231012

Claims (20)

1. 1. A method of preparing a sample for an assay for rapid quantification of a biological contaminant, the method comprising: Adding the sample to a vessel; Inserting the vessel into a centrifuge; Centrifuging the sample to produce a first supernatant and a first precipitate; Discarding the first supernatant; dissolving the first precipitate in a first volume of buffer to produce a first suspension volume; Centrifuging the first suspension volume to produce a second supernatant and a second precipitate; discarding the second supernatant; dissolving the second precipitate in a second volume of the buffer to produce a final sample suspension, and The final sample suspension was added to an assay for rapid quantification of yeast.
2. 2. The method of claim 1, wherein the sample is juice.
3. 3. The method of claim 1 or 2, wherein the sample is one or more of a stone fruit juice or berry fruit juice.
4. 4. A method according to claim 3 wherein the sample comprises at least one of apple juice, pear juice or quince juice.
5. 5. The method of claim 3, wherein the sample comprises at least one of blueberry juice, raspberry juice, medlar juice, brazil berry juice, wild cherry juice, and strawberry juice or cranberry juice.
6. 6. The method of any one of claims 1 to 5, further comprising diluting the sample.
7. 7. The method of claim 6, wherein diluting the sample is performed at least one of prior to adding the sample to a receiver, prior to inserting the receiver into a centrifuge vessel, and/or prior to centrifuging the sample.
8. 8. The method of claim 6, wherein the sample is diluted in the buffer at a ratio selected from the group consisting of 1:1000, 1:500, 1:100, 1:50, 1:20, 1:10, 1:5, 1:1, 5:1, 10:1, 20:1, 50:1, 100:1, or 1000:1.
9. 9. The method of claim 8, wherein the sample is diluted in the buffer at a ratio selected from the group consisting of 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, and 15:1.
10. 10. The method of claim 6 wherein the sample comprises citrus juice.
11. 11. The method of claim 10, wherein the sample comprises at least one of orange juice, lemon juice, lime juice, grapefruit juice, and pineapple juice.
12. 12. The method of any one of claims 1 to 11, wherein the buffer is selected from the group consisting of a calcium buffer, a carbonic acid buffer, a hydroxymethyl buffer, a nitrate buffer, a phosphate buffer, and a sulfonic acid buffer.
13. 13. The method of any one of claims 1 to 12, wherein the assay is an adenosine triphosphate assay.
14. 14. The method of claim 13, wherein the assay is a luciferase system.
15. 15. The method of claim 13, further comprising incubating the final sample suspension in the assay.
16. 16. The method of claim 15, wherein culturing the final sample suspension comprises a culture period.
17. 17. The method of claim 16, wherein the incubation period is selected from the group consisting of 1 minute, 2 minutes, 3 minutes, 5 minutes, 10 minutes, 12 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes.
18. 18. The method of claim 15, further comprising agitating the final sample suspension.
19. 19. The method of claim 18, wherein agitating the final sample suspension comprises vortexing the final sample suspension.
20. 20. The method of claim 18, wherein agitating the final sample suspension is performed at least one of prior to and/or concurrent with culturing the final sample suspension in the assay.

Description

Method and system for rapid detection of yeast Cross Reference to Related Applications The present application is filed as PCT international patent application and claims priority from U.S. provisional patent application serial No. 63/589,831 filed on 10 months 12 of 2023, the subject matter of which is hereby incorporated by reference in its entirety. Background The fruit juices are transported over long distances from suppliers to processing plants by tankers, making them susceptible to spoilage by microorganisms, especially during the warmer months. The external high temperature causes the internal temperature of the tanker to rise, thereby creating conditions for the proliferation of microorganisms. Each year, this problem has an impact on the juice industry, affecting inventory and resulting in economic losses. Conventional microbiological plating methods for counting yeasts in fruit juices typically take several days to obtain results. This time frame is too long considering that the juice needs to be accepted by the processing plant and processed quickly or rejected entirely in case the product is spoiled during transport. Other methods include assessing the taste, smell and appearance of juice, but this provides limited information about the microbiological state of the product. Disclosure of Invention The examples presented herein relate to a method of preparing a sample for an assay for rapid quantification of biological contaminants. The method includes adding a sample to a vessel and inserting the vessel into a centrifuge. The method further includes centrifuging the sample to produce a first supernatant and a first precipitate, discarding the first supernatant, and dissolving the first precipitate in a first volume of buffer to produce a first suspension volume. The method further includes centrifuging the first suspension volume to produce a second supernatant and a second precipitate, discarding the second supernatant, and dissolving the second precipitate in a second volume of buffer to produce a final sample suspension. The method further comprises adding the final sample suspension to an assay for rapid quantification of microorganisms. In other embodiments herein, the sample is a juice. In still other embodiments herein, the sample is one or more of a stone fruit juice or berry fruit juice. In other embodiments herein, the sample comprises at least one of apple juice, pear juice, or quince juice. In other embodiments herein, the sample comprises blueberry juice, raspberry juice, medlar juice, brazil berry juice, wild cherry juice, and at least one of strawberry juice or cranberry juice. In other embodiments herein, the method further comprises diluting the sample. In further embodiments herein, diluting the sample is performed at least one of prior to adding the sample to the receptacle, prior to inserting the receptacle into a centrifugation vessel, and/or prior to centrifuging the sample. In other further embodiments herein, the sample is diluted in buffer at a ratio selected from the group consisting of 1:1000, 1:500, 1:100, 1:50, 1:20, 1:10, 1:5, 1:1, 5:1, 10:1, 20:1, 50:1, 100:1, or 1000:1. In yet further embodiments herein, the sample is diluted in buffer at a ratio selected from the group consisting of 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, and 15:1. In other additional embodiments herein, the sample comprises citrus juice. In yet further embodiments herein, the sample comprises at least one of orange juice, lemon juice, lime juice, grapefruit juice, and pineapple juice. In other embodiments herein, the buffer is selected from the group consisting of a calcium buffer, a carbonic acid buffer, a hydroxymethyl buffer, a nitrate buffer, a phosphate buffer, and a sulfonic acid buffer. In other embodiments herein, the assay is an adenosine triphosphate assay. In other additional embodiments herein, the assay is a luciferase system. In other embodiments herein, the method further comprises incubating the final sample suspension in the assay. In further embodiments herein, culturing the final sample suspension comprises a culturing period. In yet further embodiments herein, the incubation period is selected from the group consisting of 1 minute, 2 minutes, 3 minutes, 5 minutes, 10 minutes, 12 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes. In other additional embodiments herein, the method further comprises agitating the final sample suspension. In other embodiments herein, agitating the final sample suspension comprises vortexing the final sample suspension. In yet further embodiments herein, agitating the final sample suspension is performed at least one of prior to and concurrent with culturing the final sample suspension in the assay. In other embodiments herein, the method further comprises reading the final suspension using a photometer to obtain a result, wherein the result is measured in Relative Light Units (RLU). In yet further embodiments