Nucleic acid amplification apparatus and method

Abstract

A nucleic acid amplification apparatus for amplifying a target nucleic acid derived from living organism, comprising: a measurement unit for amplifying the target nucleic acid in a measuring sample prepared from the living organism, and measuring a product of the amplification of the target nucleic acid; a measurement value obtaining unit for obtaining a measurement value related to an amount of the product of the amplification; and a judging unit for judging whether amplification inhibition of the target nucleic acid occur or not based on a first measurement value and a second measurement value, the first measurement value obtained from a first measurement sample and the second measurement value obtained from a second measurement sample having a difference dilution ratio from the first measurement sample.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a perspective illustration showing whole composition of a gene amplification and analysis system by one embodiment according to the present invention.

FIG. 2 is a perspective illustration showing whole composition of a gene amplification measuring apparatus of the gene amplification and analysis system by one embodiment shown in FIG. 1.

FIG. 3 is a schematic plan view of FIG. 2.

FIG. 4 is a drawing showing data browser screen displayed on the display unit of personal computer composing the gene amplification and analysis system by one embodiment shown in FIG. 1.

FIG. 5 is a drawing showing relationship between measurement results of sample specimen, measurement results of dilution sample, and existence or nonexistence of target gene.

FIG. 6 is a graph showing relationship between dilution rate and concentration of sample specimen that caused amplification inhibition.

FIG. 7 is a drawing showing workload list screen displayed on the display unit of the personal computer composing the gene amplification and analysis system by one embodiment shown in FIG. 1.

FIG. 8 is a drawing showing analytical curve display screen displayed on the display unit of the personal computer composing the gene amplification and analysis system by one embodiment shown in FIG. 1.

FIG. 9 is a schematic plan view showing variation of the gene amplification measuring apparatus of the gene amplification and analysis system by one embodiment shown in FIG. 1.

FIG. 10 is a flowchart showing amplification inhibition judgment flow of CPU of the personal computer of the gene amplification and analysis system by one embodiment shown in FIG. 1. FIG. 1 is a plan view showing whole composition of an immune analysis apparatus equipped with a pipette chip supplying apparatus by one embodiment according to the present invention.

Claims

1. A nucleic acid amplification apparatus for amplifying a target nucleic acid derived from living organism, comprising: a measurement unit for amplifying the target nucleic acid in a measuring sample prepared from the living organism, and measuring a product of the amplification of the target nucleic acid;a measurement value obtaining unit for obtaining a measurement value related to an amount of the product of the amplification; anda judging unit for judging whether amplification inhibition of the target nucleic acid occur or not based on a first measurement value and a second measurement value, the first measurement value obtained from a first measurement sample and the second measurement value obtained from a second measurement sample having a difference dilution ratio from the first measurement sample.

2. The nucleic acid amplification apparatus according to claim 1, further comprising an output unit for outputting information related to the judgment result by the judging unit.

3. The nucleic acid amplification apparatus according to claim 1, wherein the measurement sample comprises a nucleic acid derived from living organism, a nucleic acid amplification enzyme and a primer for the target nucleic acid.

4. The nucleic acid amplification apparatus according to claim 1, further comprising an analysis unit for judging whether the target nucleic acid in the living organism excessively express or not based on comparison between a threshold value and the first measurement value when the judging unit judges no occurrence of amplification inhibition.

5. The nucleic acid amplification apparatus according to claim 4, wherein the analysis unit judges whether an the target nucleic acid in the living organism excessively express or not based on comparison between a second threshold value and the second measurement value when the judging unit judges occurrence of amplification inhibition.

6. The nucleic acid amplification apparatus according to claim 1, wherein the judging unit judges occurrence of amplification inhibition when an amount of the second product based on the second measurement value is larger than that of the first product based on the first measurement value.

7. The nucleic acid amplification apparatus according to claim 1, wherein the second measurement sample have higher dilution ratio than that of the first measurement sample.

8. The nucleic acid amplification apparatus according to claim 4, further comprising a output unit for outputting information relating to the judgment result by the analysis unit.

9. The nucleic acid amplification apparatus according to claim 4, further comprising a output unit for outputting positive or negative based on the judgment result relating to the excess expression.

10. The nucleic acid amplification apparatus according to claim 1, further comprising: a sample setting part for setting a first container and a second container, the first container accommodating the first measuring sample and the second container accommodating the second measuring sample; anda dispensing unit for dispensing the first measuring sample from the first container to a first reaction container in the measurement unit and dispensing the second measuring sample from the second container to a second reaction container in the measurement unit.

11. The nucleic acid amplification apparatus according to claim 1, further comprising: a sample setting part for setting a first container and a second container, the first container accommodating a pretreatment sample which is prepared by the living organism and a pretreatment liquid and contains solubilized nucleic acid in the pretreatment liquid, and the second container accommodating a dilution liquid; anda dispensing unit for dispensing the pretreatment sample from the first container to a first reaction container and a second reaction container in the measurement unit and dispensing the dilution liquid from the second container to the second reaction container.

12. The nucleic acid amplification apparatus according to claim 11, wherein the pretreatment sample is a sample in which nucleic acid is not purified.

13. A nucleic acid amplification method for amplifying a target nucleic acid derived from living organism, comprising steps of: amplifying the target nucleic acid in a first measuring sample prepared from the living organism;amplifying the target nucleic acid in a second measuring sample prepared from the living organism and had a difference dilution ratio from the first measurement sample;measuring a first product of the amplification of the target nucleic acid of the first measurement sample;measuring a second product of the amplification of the target nucleic acid of the second measurement sample;obtaining a first measurement value related to an amount of the first product of the amplification;obtaining a second measurement value related to an amount of the second product of the amplification; andjudging whether amplification inhibition of the target nucleic acid occur or not based on the first measurement value and the second measurement value.

14. The nucleic acid amplification method according to claim 13, further comprising the step of: outputting information relating to the judging result.

15. The nucleic acid amplification method according to claim 13, wherein the first measurement sample comprises a nucleic acid derived from living organism, a nucleic acid amplification enzyme and a primer for the target nucleic acid, and the second measurement sample comprises a nucleic acid derived from living organism, a nucleic acid amplification enzyme, a primer for the target nucleic acid and a dilution liquid.

16. The nucleic acid amplification method according to claim 13, further comprising the step of: judging whether the target nucleic acid in the living organism excessively express or not based on comparison between a threshold value and the first measurement value when the judging result is no occurrence of amplification inhibition.

17. The nucleic acid amplification method according to claim 13, further comprising the step of: judging whether the target nucleic acid in the living organism excessively express or not based on comparison between a second threshold value and the second measurement value when the judging result is occurrence of amplification inhibition.

18. The nucleic acid amplification method according to claim 13, wherein the judging step includes the judging of occurrence of amplification inhibition when an amount of the second product based on the second measurement value is larger than that of the first product based on the first measurement value.

19. The nucleic acid amplification method according to claim 13, wherein the second measurement sample have higher dilution ratio than that of the first measurement sample.

20. The nucleic acid amplification method according to claim 16, further comprising step of: outputting positive or negative based on the judgment result relating to the excess expression.