Sample processing apparatus and method

Patent metadata

Field	Value
Publication number	US-12065698-B2
Application number	US-202117498308-A
Country	US
Kind code	B2
Filing date	Oct 11, 2021
Priority date	Oct 2, 2009
Publication date	Aug 20, 2024
Grant date	Aug 20, 2024

Abstract

The invention relates to a sample processing apparatus comprising a holder for a microtiter plate comprising a plurality of microwells, optical measurement unit for measuring optical responses of samples dosed to the microwells, and a computing unit configured to analyze the optical responses in order to detect dosing failures in said plurality of microwells, and if a dosing failure has been detected in one or more of the microwells, to communicate the existence of the dosing failure to a user of the apparatus through signaling means or to store data indicative of the dosing failure to data storage means for further use. In particular, the invention relates to detecting dosing failures in before, during and after a PCR process.

First claim

The invention claimed is:

1. A method of preparing a reaction mixture for Polymerase Chain Reaction (PCR) assay, comprising
providing a first reagent solution comprising at least one substance required for performing said assay,
providing a second reagent solution comprising at least one other substance required for performing said assay,
mixing the first and second reagent solutions,
providing a third reagent solution comprising at least one further substance required for performing said assay,
mixing the third reagent solution with the first and second reagent solutions
for providing a mixed solution to be subjected to the PCR process, characterized in that
the first reagent solution contains a first colorant providing the first reagent solution a first color,
the second reagent solution contains a second colorant providing the solution a second color different from the first color,
said mixing of the first and second reagent solutions yields a mixed solution having, due to said first and second colorants, a third color different from the first and second colors, and wherein the third color is detected visually prior to subjecting the mixed solution to the PCR process,
the third reagent solution contains a third colorant providing the solution a fourth color different from the first, second, and third colors,
said mixing of the first, second, and third reagent solutions yields a mixed solution having, due to said first, second, and third colorants, a fifth color, different from the first, second, third, and fourth colors,
the first or second reagent solution comprises an Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye.

2. A solution set for Polymerase Chain Reaction (PCR) assay, comprising
a first reagent solution comprising at least one substance required for performing said PCR assay,
a second reagent solution comprising at least one other substance required for performing said PCR assay,
a third reagent solution comprising at least one further substance required for performing said assay,
characterized in that
the first reagent solution is provided with a first colorant having a first color,
the second reagent solution is provided with a second colorant having a second color different from the first color,
the first and second reagent solutions are capable of forming, on mixing, a first mixed solution having, due to said first and second colorants, a third color different from the first and second colors, wherein the colors are capable of being detected visually,
the third reagent solution contains a third colorant providing the solution a fourth color different from the first, second, and third colors,
the first, second, and third reagent solutions are capable of forming, on mixing, a second mixed solution having, due to said first, second, and third colorants, a fifth color different from the first, second, third, and fourth colors,
the first or second reagent solution comprises an Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye.

3. The solution set according to claim 2 , characterized in that at least one of said reagent solutions is a sample solution for receiving a template to be amplified.

4. The solution set according to claim 2 , characterized in that at least one of the reagent solutions contains one or more of the following: polymerase, primer, ions or dNTPs or fluorescent qPCR dye or probe, preferably being a PCR master mix, in particular a qPCR master mix.

5. The solution set according to claim 2 , characterized in that at least one of the reagent solutions is an elution buffer or a dilution buffer.

6. The solution set according to claim 2 , characterized in that said first and/or second colorants are selected from the group of: Quinoline yellow, Xylene cyanol, Brilliant Blue, Patent Blue, Indigocarmine, Acid Red 1, m-Cresol Purple, Cresol Red, Neutral Red, Bromocresol Green, Acid Violet 5, Bromo phenol blue, and Orange G.

7. The solution set according to claim 2 , characterized in that the concentration of said colorants in respective reagent solutions corresponds to absorbances of the said solutions at their maximum absorption wavelengths of 0.01-0.5, preferably 0.03-0.15 with 1 mm light path, when diluted to the desired PCR processing concentration.

8. The solution set according to claim 2 , characterized in that the first reagent solution and/or the second reagent solution is a concentrate.

9. The solution set according to claim 2 , characterized in that the first reagent solution and the second reagent solution are capable of forming a transparent or translucent mixed solution suitable for qPCR.

10. The solution set according to claim 2 , characterized in that the first reagent solution is selected from: polymerase solution, polymerase buffer; and the second reagent solution is selected from: primers, probe and sample.

11. The solution set according to claim 2 , wherein the absorbance peaks of any of said colorants do not overlap with the emission or excitation wavelength of the Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye.

12. The solution set according to claim 2 , characterized in that the total absorbance of the mixed solution at the emission or excitation wavelength of the Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye is less than 0.05, preferably less than 0.03, in particular less than 0.1 (1 mm light path).

13. Use of the method according to claim 1 , for preparing a reaction mixture for quantitative PCR.

14. Use of the solution set according to claim 1 , for preparing a reaction mixture for quantitative PCR.

Assignees

• Thermo Fisher Scientific Baltics Uab

Inventors

• Tuunanen Jukka
• Kurkela Jaako
• Eklin Katja

Classifications

• G01N21/78

  producing a change of colour · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• G01N2021/6439

  with indicators, stains, dyes, tags, labels, marks · CPC title

• G01N21/29

  using visual detection (G01N21/31 takes precedence) · CPC title