Method, device and system for testing drug sensitivity

Patent metadata

Field	Value
Publication number	US-10619182-B2
Application number	US-201415526388-A
Country	US
Kind code	B2
Filing date	Dec 26, 2014
Priority date	Dec 26, 2014
Publication date	Apr 14, 2020
Grant date	Apr 14, 2020

Abstract

When bacteria growth is determined in the related art using an ATP method, the bacteria growth is determined, based on an increase or a decrease in live bacteria ATP with the lapse of a culture time. Accordingly, it is necessary to measure the live bacteria ATP multiple times while antimicrobial susceptibility culture is carried out. It takes time and labor in sample preparation for each measurement, and it is difficult to quickly obtain an antimicrobial susceptibility result. Therefore, according to the present invention, the presence or absence of antimicrobial susceptibility of bacteria is determined, based on an ATP luminescence amount derived from dead bacteria in a culture liquid.

First claim

The invention claimed is:

1. An antimicrobial susceptibility test system comprising:
a culture vessel that cultures a bacterial cell in a culture liquid;
a dispenser that dispenses an antimicrobial into a distribution container to which the culture liquid containing the bacterial cell is distributed and dispenses the culture liquid from the distribution container to a filtering container disposed above a first luminescence measurement container and to a second luminescence measurement container, the filtering container having a filter in a bottom portion;
wherein the filter filters the culture liquid in the filtering container and obtains a filtrate into the first luminescence measurement container; and
a test device that has a luminescence detector and a determination unit, the luminescence detector measuring ATP luminescence of the filtrate in the first luminescence measurement container and the culture liquid in the second luminescence measurement container, and the determination unit being programmed to determine the presence or absence of antimicrobial susceptibility of the bacteria, based on an ATP luminescence amount derived from dead bacteria in the culture liquid,
wherein the luminescence detector measures an ATP luminescence amount derived from live bacteria in the culture liquid in the second luminescence measurement container and the ATP luminescence amount derived from the dead bacteria in the filtrate in the first luminescence measurement container, and
wherein the determination unit is programmed to determine the presence or absence of antimicrobial susceptibility of the bacteria, based on a luminescence amount ratio between the ATP luminescence amount derived from the live bacteria and the ATP luminescence amount derived from the dead bacteria.

2. The antimicrobial susceptibility test system according to claim 1 ,
wherein the determination unit is programmed to perform the determination by regarding a luminescence amount of free ATP included in the filtrate as the ATP luminescence amount derived from the dead bacteria.

3. The antimicrobial susceptibility test system according to claim 1 , further comprising:
a database storage that stores a threshold for the determination,
wherein the determination unit is programmed to refer to the threshold and determine the presence or absence of antimicrobial susceptibility of the bacteria, based on a comparison result of a luminescence amount ratio between the ATP luminescence amount derived from the live bacteria and the ATP luminescence amount derived from the dead bacteria and the threshold.

4. The antimicrobial susceptibility test system according to claim 3 ,
wherein the determination unit is programmed to refer to the threshold, and determine that the antimicrobial susceptibility of the bacteria is present in a case where the luminescence amount ratio is greater than the threshold.

5. The antimicrobial susceptibility test system according to claim 3 ,
wherein the database storage stores the threshold, based on the luminescence amount ratio obtained by the determination in the past.

6. The antimicrobial susceptibility test system according to claim 3 ,
wherein the determination unit is programmed to set the ATP luminescence amount derived from the dead bacteria to x and the ATP luminescence amount derived from the live bacteria to y, calculate a value of x/y, and determine the presence or absence of antimicrobial susceptibility of the bacteria by referring to a value of x/y for each type of the bacteria calculated in a past and stored in the database storage.

7. The antimicrobial susceptibility test system according to claim 1 ,
wherein the luminescence detector performs the measurement at constant incubation time intervals.

8. The antimicrobial susceptibility test system according to claim 1 ,
wherein the filter performs the filtration by a centrifuge.

9. The antimicrobial susceptibility test system according to claim 1 ,
wherein the distribution container is a micro well plate to which a plurality of types of the antimicrobial is distributed.

10. The antimicrobial susceptibility test system according to claim 1 ,
wherein the distribution container is a micro well plate to which a plurality of concentrations of the antimicrobial is distributed.

11. An antimicrobial susceptibility test system comprising:
a culture vessel to culture a bacterial cell in a culture liquid;
a dispenser to dispense an antimicrobial into a distribution container to which the culture liquid containing the bacterial cell is distributed and dispense the culture liquid from the distribution container to a filtering container disposed above a first luminescence measurement container and to a second luminescence measurement container, the filtering container having a filter in a bottom portion to filter the culture liquid in the filtering container and obtain a filtrate into the first luminescence measurement container; and
a test device that has a luminescence detector and a determination unit, the luminescence detector measuring ATP luminescence of the filtrate in the first luminescence measurement container and the culture liquid in the second luminescence measurement container, the determination unit determining the presence or absence of antimicrobial susceptibility of the bacteria based on an ATP luminescence amount derived from dead bacteria in the culture liquid;
wherein the luminescence detector is configured to measure an ATP luminescence amount derived from live bacteria in the culture liquid in the second luminescence measurement container and the ATP luminescence amount derived from the dead bacteria in the filtrate in the first luminescence measurement container; and
wherein the determination unit is programmed to determine the presence or absence of antimicrobial susceptibility of the bacteria, based on a luminescence amount ratio between the ATP luminescence amount derived from the live bacteria and the ATP luminescence amount derived from the dead bacteria in the culture liquid.

12. The antimicrobial susceptibility test system according to claim 11 ,
wherein the determination unit is programmed to perform the determination by regarding a luminescence amount of free ATP included in the filtrate as the ATP luminescence amount derived from the dead bacteria.

13. The antimicrobial susceptibility test system according to claim 11 , further comprising:
a computer memory that stores a threshold for the determination,
wherein the determination unit is programmed to refer to the threshold and determine the presence or absence of antimicrobial susceptibility of the bacteria, based on a comparison result of a luminescence amount ratio between the ATP luminescence amount derived from the live bacteria and the ATP luminescence amount derived from the dead bacteria and the threshold.

14. The antimicrobial susceptibility test system according to claim 13 ,
wherein the determination unit is programmed to refer to the threshold, and determine that the antimicrobial susceptibility of the bacteria is present in a case where the luminescence amount ratio is greater than the threshold.

15. The antimicrobial susceptibility test system according to claim 13 ,
wherein the computer memory stores the threshold, based on the luminescence amount ratio obtained by the determination in the past.

16. The antimicrobial susceptibility test system according to claim 13 ,
wherein the determination unit is programmed to set the ATP luminescence amount derived from the dead bacteria to x and the ATP luminescence amount derived from the live bacteria

Assignees

• Hitachi Ltd

Inventors

• Tada Hiroko
• Noda Hideyuki
• Niimi Hideki
• Kitajima Isao

Classifications

• C12Q1/66

  involving luciferase · CPC title

• C12M1/34

  Measuring or testing with condition measuring or sensing means, e.g. colony counters · CPC title

• C12Q1/18 Primary

  Testing for antimicrobial activity of a material · CPC title

• C12Q1/008 Primary

  for determining co-enzymes or co-factors, e.g. NAD, ATP · CPC title