CA2631425A1 - Controls for nucleic acid testing - Google Patents
Controls for nucleic acid testing Download PDFInfo
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- CA2631425A1 CA2631425A1 CA002631425A CA2631425A CA2631425A1 CA 2631425 A1 CA2631425 A1 CA 2631425A1 CA 002631425 A CA002631425 A CA 002631425A CA 2631425 A CA2631425 A CA 2631425A CA 2631425 A1 CA2631425 A1 CA 2631425A1
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- nucleic acid
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- biomolecule
- internal control
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
Abstract
The present invention is related to a method for detecting a target biomolecule in test sample by adding an internal control biomolecule to the test sample; to a negative control sample, to a positive control sample and to a reagent control sample or adding an internal control biomolecule to the test sample, to a negative control sample, to a positive control sample comprising the target biomolecule and providing a reagent control sample comprising the target biomolecule, determining in each sample a signal, and verifying the signal thereby detecting the target biomolecule. The invention is also related to a method for verifying the determination of a signal indicating the presence of a target biomolecule. The invention is further related to a method for detecting the presence or the absence of a member of a group of target nucleic acids in a sample and a method for verifying the determination of a signal indicating the presence of a member of a group of target nucleic acids.
Uses and kits are considered as well.
Uses and kits are considered as well.
Claims (43)
1. A method for detecting the presence or the absence of a target biomolecule in a sample suspected to comprise the target biomolecule comprising the following steps:
a) wherein this step consists of step a1) or a2) a1) adding an internal control biomolecule - to the sample suspected to comprise the target biomolecule, and - to a negative control sample not comprising the target biomolecule, - to a positive control sample comprising the target biomolecule, and - to a reagent control sample comprising the target biomolecule a2) adding an internal control biomolecule - to the sample suspected to comprise the target biomolecule, and - to a negative control sample not comprising the target biomolecule, and - to a positive control sample comprising the target biomolecule, and providing a reagent control sample comprising the target biomolecule b) optionally purifying the biomolecules from the samples of step a) to obtain samples comprising the purified biomolecules, c) determining in each sample obtained in step a) or b) the presence or the absence of a signal of the internal control biomolecule and of the target biomolecule, d) verifying the presence or absence of the signal of the target biomolecule in the sample suspected to comprise the target biomolecule by:
- checking the sample suspected to comprise the target biomolecule for the presence of a signal for the target biomolecule independently from the presence of a signal of the internal control biomolecule or checking for the presence of a signal of the internal control biomolecule in the case of an absence of a signal for the target biomolecule, - checking the negative control sample for the presence of a signal of the internal control biomolecule and for the absence of a signal of the target biomolecule, - checking the positive control sample for the presence of a signal of the target biomolecule and for the presence of a signal of the internal control biomolecule, and - checking the reagent control sample for the presence of a signal for the target biomolecule in step d) of the method or checking the reagent control sample for the presence of a signal for the target biomolecule and optionally for the internal control biomolecule e) detecting the presence or the absence of the target biomolecule whereby the presence or absence of the signals for the target biomolecule and the internal control biomolecule determined in step c) and verified in step d) indicate the presence or the absence of the target biomolecule in the test sample.
a) wherein this step consists of step a1) or a2) a1) adding an internal control biomolecule - to the sample suspected to comprise the target biomolecule, and - to a negative control sample not comprising the target biomolecule, - to a positive control sample comprising the target biomolecule, and - to a reagent control sample comprising the target biomolecule a2) adding an internal control biomolecule - to the sample suspected to comprise the target biomolecule, and - to a negative control sample not comprising the target biomolecule, and - to a positive control sample comprising the target biomolecule, and providing a reagent control sample comprising the target biomolecule b) optionally purifying the biomolecules from the samples of step a) to obtain samples comprising the purified biomolecules, c) determining in each sample obtained in step a) or b) the presence or the absence of a signal of the internal control biomolecule and of the target biomolecule, d) verifying the presence or absence of the signal of the target biomolecule in the sample suspected to comprise the target biomolecule by:
- checking the sample suspected to comprise the target biomolecule for the presence of a signal for the target biomolecule independently from the presence of a signal of the internal control biomolecule or checking for the presence of a signal of the internal control biomolecule in the case of an absence of a signal for the target biomolecule, - checking the negative control sample for the presence of a signal of the internal control biomolecule and for the absence of a signal of the target biomolecule, - checking the positive control sample for the presence of a signal of the target biomolecule and for the presence of a signal of the internal control biomolecule, and - checking the reagent control sample for the presence of a signal for the target biomolecule in step d) of the method or checking the reagent control sample for the presence of a signal for the target biomolecule and optionally for the internal control biomolecule e) detecting the presence or the absence of the target biomolecule whereby the presence or absence of the signals for the target biomolecule and the internal control biomolecule determined in step c) and verified in step d) indicate the presence or the absence of the target biomolecule in the test sample.
2. The method according to claim 1 wherein the negative control sample is a solution comprising a salt and a buffer substance.
3. The method according to any of the claims 1 to 2 wherein the internal control biomolecule comprises a part of the target biomolecule.
4. The method according to any of the claims 1 to 3 wherein the reagent control sample and/ or the positive control sample is a solution comprising the target biomolecule or a part thereof.
5. The method according to any of the claims 1 to 4 wherein the positive control sample comprises a virus, a microorganism or a cell comprising the target biomolecule.
6. The method according to any of the claims 1 to 5 wherein the target biomolecule is a group of target biomolecules.
7. The method according to any of the claims 1 to 6 wherein the biomolecule is a nucleic acid.
8. The method according to claim 7 wherein the target nucleic acid and the internal control nucleic acid is amplified before step c).
9. The method according to any of the claims 7 to 8 wherein the signal of the target nucleic acid or of the internal control nucleic acid is a fluorescence signal.
10. The method according to claim 9 wherein the fluorescence signal is generated by a label attached to a probe that hybridizes to the target nucleic acid or the internal control nucleic acid.
11. The method according to claim 10 wherein the fluorescence signal is generated by a pair of probes hybridizing to the respective nucleic acid wherein the members of said pair of probes hybridize to the respective nucleic acid within no more than five nucleotides distance to one another, wherein the first probe of said pair of probes is labeled with a donor fluorescent label and wherein the second probe of said pair of probes is labeled with a corresponding acceptor fluorescent label and wherein the hybridization of the first and second probe to the respective nucleic acid brings them into in a resonance energy transfer relationship.
12. The method according to claim 11 wherein step c) comprises the following substeps c1) adding to the samples obtained in step a) or b) - a pair of primers hybridizing to the internal control nucleic acid and the target nucleic acid or two pairs of primers the first one hybridizing to the internal control nucleic acid and the second one hybridizing to the target nucleic acid, - a first pair of probes hybridizing to the internal control nucleic acid wherein the members of said first pair of probes hybridize to the internal control nucleic acid within no more than five nucleotides distance to one another, wherein the first probe of said first pair of probes is labeled with a first donor fluorescent label and wherein the second probe of said first pair of probes is labeled with a corresponding first acceptor fluorescent label and wherein the hybridization of the first and second probe to the internal control nucleic acid brings them into in a resonance energy transfer relationship;
- a second pair of probes hybridizing to the target nucleic acid wherein the members of the second pair of probes hybridize to the target nucleic acid within no more than five nucleotides distance to one another, wherein the first probe of said second pair of probes is labeled with a second donor fluorescent label and wherein the second probe of said second pair of probes is labeled with a corresponding second acceptor fluorescent label and wherein the hybridization of the first and second probe to the target nucleic acid brings them into in a resonance energy transfer relationship; and - a thermostable nucleic acid polymerase and reagents necessary for amplifying the internal control nucleic acid and the target nucleic acid, c2) amplifying in the samples the internal control nucleic acid and the target nucleic acid if present in the respective sample, c3) determining in each sample separately the presence or absence of a fluorescence signal of the internal control nucleic acid and of the target nucleic acid as a function of the temperature of the respective sample, whereby the fluorescence signal specific for the internal control nucleic acid is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the first pair of probes and the first acceptor fluorescent label of the second probe of the first pair of probes, and - whereby the fluorescence signal specific for the target nucleic acid is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the second pair of probes and the first acceptor fluorescent label of the second probe of the second pair of probes.
- a second pair of probes hybridizing to the target nucleic acid wherein the members of the second pair of probes hybridize to the target nucleic acid within no more than five nucleotides distance to one another, wherein the first probe of said second pair of probes is labeled with a second donor fluorescent label and wherein the second probe of said second pair of probes is labeled with a corresponding second acceptor fluorescent label and wherein the hybridization of the first and second probe to the target nucleic acid brings them into in a resonance energy transfer relationship; and - a thermostable nucleic acid polymerase and reagents necessary for amplifying the internal control nucleic acid and the target nucleic acid, c2) amplifying in the samples the internal control nucleic acid and the target nucleic acid if present in the respective sample, c3) determining in each sample separately the presence or absence of a fluorescence signal of the internal control nucleic acid and of the target nucleic acid as a function of the temperature of the respective sample, whereby the fluorescence signal specific for the internal control nucleic acid is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the first pair of probes and the first acceptor fluorescent label of the second probe of the first pair of probes, and - whereby the fluorescence signal specific for the target nucleic acid is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the second pair of probes and the first acceptor fluorescent label of the second probe of the second pair of probes.
13. The method according to any of the claims 7 to 12 wherein the target nucleic acid is a group of target nucleic acids and the pair of probes is a group of pairs of probes whereby each pair of probes hybridizes to a member of the group of target nucleic acids.
14. The method according to any of the claims 10 to 12 wherein the label is a fluorescein dye, a rhodamine dye, a cyanine dye, or a coumarin dye.
15. The method according to any of the claims 7 to 14 wherein the nucleic acid sequence of target nucleic acid is a nucleic acid sequence specific for a microorganism, a cell or a virus.
16. The method according to claim 15 wherein the microorganism is a gram-positive or a gram-negative microorganism or a fungi.
17. The method according to claim 16 wherein a) the gram-positive microorganism is Proteus mirabilis, Serratia marcescens, Acinetobacter baumannii, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Pseudomonas aeruginosa, or Stenotrophomonas maltophilia, b) the gram-negative microorganism is a Staphylococcus spp., Enterococcus faecium or faecalis or a Streptococcus spp., or c) the fungi is Candida albicans, Aspergillus fumigatus, Candida krusei, Candida glabrata, Candida parapsilosis, or Candida tropicalis.
18. A method for verifying the determination of a signal indicating the presence of a target biomolecule comprising the steps of a) providing - a sample suspected to comprise the target biomolecule and comprising an internal control biomolecule, and - a negative control sample comprising an internal control biomolecule and not comprising the target biomolecule, and - a positive control sample comprising the target biomolecule and comprising an internal control biomolecule - a reagent control sample comprising the target biomolecule and optionally comprising an internal control biomolecule b) determining in each sample the signal of the internal control biomolecule and of the target biomolecule, c) verifying the presence of the signal of the target biomolecule in the test sample indicating the presence of the target biomolecule in the test sample by:
- checking the sample suspected to comprise the target biomolecule for the presence of a signal for the target biomolecule independently from the presence of a signal of the internal control biomolecule or checking for the presence of a signal of the internal control biomolecule in the case of an absence of a signal for the target biomolecule, - checking the negative control sample for the presence of a signal of the internal control biomolecule and for the absence of a signal of the target biomolecule, - checking the positive control sample for the presence of a signal of the target biomolecule and for the presence of a signal of the internal control biomolecule, and - checking the reagent control sample for the presence of a signal for the target biomolecule and optionally for the internal control biomolecule.
- checking the sample suspected to comprise the target biomolecule for the presence of a signal for the target biomolecule independently from the presence of a signal of the internal control biomolecule or checking for the presence of a signal of the internal control biomolecule in the case of an absence of a signal for the target biomolecule, - checking the negative control sample for the presence of a signal of the internal control biomolecule and for the absence of a signal of the target biomolecule, - checking the positive control sample for the presence of a signal of the target biomolecule and for the presence of a signal of the internal control biomolecule, and - checking the reagent control sample for the presence of a signal for the target biomolecule and optionally for the internal control biomolecule.
19. The method according to claim 18 - wherein the internal control biomolecule comprises a part of the target biomolecule, - wherein the positive control sample comprises a virus, a microorganism or a cell containing the target biomolecule, or wherein the positive control sample is a solution comprising the target biomolecule or a part thereof, or - wherein the reagent control sample is a solution comprising the target biomolecule or a part thereof.
20. The method according to any of the claims 18 to 19 wherein the target biomolecule is a group of target biomolecules.
21. The method according to any of the claims 18 to 20 wherein the biomolecule is a nucleic acid.
22. The method according to claim 21 wherein the target nucleic acid and the internal control nucleic acid is amplified before step b).
23. The method according to any of the claims 21 to 22 wherein the signal of the target nucleic acid or of the internal control nucleic acid is a fluorescence signal.
24. The method according to claim 23 wherein the fluorescence signal is generated by a label attached to a probe that hybridizes to the target nucleic acid or the internal control nucleic acid.
25. A method for detecting the presence or the absence of a member of a group of target nucleic acids in a sample suspected to comprise the member of a group of target nucleic acids by:
a) adding an internal control nucleic acid - to the sample suspected to comprise a member of the group of target nucleic acids, and - to a negative control sample not comprising a member of the group of target nucleic acids, and - to a positive control sample comprising a member of the group of target nucleic acids, b) providing a reagent control sample comprising the group of target nucleic acids and optionally an internal control nucleic acid, c) optionally purifying the nucleic acids from the samples of step a) and/ or b) to obtain samples comprising the purified nucleic acids, d) determining in each sample obtained in the steps a) and b) or in the steps b) and c) the presence or absence of a signal of the internal control nucleic acid and of a signal of a member of the group of target nucleic acids, e) verifying the presence or the absence of the signal of the member of the group of target nucleic acids in the sample suspected to comprise a member of the group of target nucleic acids by:
- checking the sample suspected to comprise a member of the group of target nucleic acids for the presence of a signal of a member of the group of target nucleic acids independently from the presence of a signal of the internal control nucleic acid or checking for the presence of a signal of the internal control nucleic acid in the case of an absence of a signal of a member of the group of target nucleic acids, - checking the negative control sample for the presence of a signal of the internal control nucleic acid and for the absence of a signal of the target nucleic acid, - checking the reagent control sample for the presence of a signal of each member of the group of target nucleic acids and optionally of the internal control nucleic acid, and - checking the positive control sample for the presence of a signal of a member of the group of target nucleic acids and for the presence of a signal of the internal control nucleic acid or, f) detecting the presence or the absence of a member of the group of target nucleic acids whereby the presence and/ or the absence of the signals for a member of the group of target nucleic acids and the internal control nucleic acid determined in step d) and verified in step e) indicate the presence or the absence of a member of the group of target nucleic acids in the sample suspected to comprise a member of the group of target nucleic acids.
a) adding an internal control nucleic acid - to the sample suspected to comprise a member of the group of target nucleic acids, and - to a negative control sample not comprising a member of the group of target nucleic acids, and - to a positive control sample comprising a member of the group of target nucleic acids, b) providing a reagent control sample comprising the group of target nucleic acids and optionally an internal control nucleic acid, c) optionally purifying the nucleic acids from the samples of step a) and/ or b) to obtain samples comprising the purified nucleic acids, d) determining in each sample obtained in the steps a) and b) or in the steps b) and c) the presence or absence of a signal of the internal control nucleic acid and of a signal of a member of the group of target nucleic acids, e) verifying the presence or the absence of the signal of the member of the group of target nucleic acids in the sample suspected to comprise a member of the group of target nucleic acids by:
- checking the sample suspected to comprise a member of the group of target nucleic acids for the presence of a signal of a member of the group of target nucleic acids independently from the presence of a signal of the internal control nucleic acid or checking for the presence of a signal of the internal control nucleic acid in the case of an absence of a signal of a member of the group of target nucleic acids, - checking the negative control sample for the presence of a signal of the internal control nucleic acid and for the absence of a signal of the target nucleic acid, - checking the reagent control sample for the presence of a signal of each member of the group of target nucleic acids and optionally of the internal control nucleic acid, and - checking the positive control sample for the presence of a signal of a member of the group of target nucleic acids and for the presence of a signal of the internal control nucleic acid or, f) detecting the presence or the absence of a member of the group of target nucleic acids whereby the presence and/ or the absence of the signals for a member of the group of target nucleic acids and the internal control nucleic acid determined in step d) and verified in step e) indicate the presence or the absence of a member of the group of target nucleic acids in the sample suspected to comprise a member of the group of target nucleic acids.
26. The method according to claim 25 - wherein the internal control biomolecule comprises a part of the target biomolecule, - wherein the positive control sample comprises a virus, a microorganism or a cell containing the target biomolecule or wherein the positive control sample is a solution comprising the target biomolecule or a part thereof, or - wherein the reagent control sample is a solution comprising the target biomolecule or a part thereof.
27. The method according to any of the claims 25 to 26 wherein a member of the group of target nucleic acids and the internal control nucleic acid is amplified before step d).
28. The method according to any of the claims 25 to 27 wherein the signal of a member of the group of target nucleic acids or of the internal control nucleic acid is a fluorescence signal.
29. The method according to claim 28 wherein the fluorescence signal is generated by a label attached to a probe that hybridizes to a member of the group of target nucleic acids or the internal control nucleic acid.
30. The method according to claim 29 wherein the fluorescence signal is generated by a member of a group of pairs of probes hybridizing to a member of the group of target nucleic acids or of a pair of probes hybridizing to the internal control nucleic acid wherein the members of each member of the group of pairs of probes hybridizing to the member of the group of target nucleic acids or the members of the pair of probes hybridizing to the internal control nucleic acid hybridize to the respective nucleic acid within no more than five nucleotides distance to one another, wherein the first probe of a pair of probes is labeled with a donor fluorescent label and wherein the second probe of a pair of probes is labeled with a corresponding acceptor fluorescent label and wherein the hybridization of the first and second probe to the member of the group of target nucleic acids or the internal control nucleic acid brings them into a resonance energy transfer relationship.
31. The method according to claim 30 wherein step d) comprises the following substeps dl ) adding to the samples obtained in step a) and b) or steps b) and c) - a pair of primers hybridizing to the internal control nucleic acid and to a member of the group of target nucleic acids or a pair of primers hybridizing to the internal control nucleic acid and a group of of pairs of primer each member thereof hybridizing to a member of the group of target nucleic acids, - a first pair of probes hybridizing to the internal control nucleic acid wherein the members of said first pair of probes hybridize to the internal control nucleic acid within no more than five nucleotides distance to one another, wherein the first probe of said first pair of probes is labeled with a first donor fluorescent label and wherein the second probe of said first pair of probes is labeled with a corresponding first acceptor fluorescent label and wherein the hybridization of the first and second probe to the internal control nucleic acid brings them into in a resonance energy transfer relationship;
- a group of pairs of probes each member hybridizing to a member of the group of target nucleic acids wherein the members of the group of pairs of probes hybridize to the respective member of the group of target nucleic acids within no more than five nucleotides distance to one another, wherein the first probe of a member of the group of pairs of probes is labeled with a second donor fluorescent label and wherein the second probe of a member of the group of pairs of probes is labeled with a corresponding second acceptor fluorescent label and wherein the hybridization of the first and second probe to the member of the group of target nucleic acids brings them into in a resonance energy transfer relationship; and - a thermostable nucleic acid polymerase and reagents necessary for amplifying the internal control nucleic acid and the group of target nucleic acids, d2) amplifying in the samples the internal control nucleic acid and the group of target nucleic acids if present in the respective sample, d3) determining in each sample separately the presence or absence of a fluorescence signal of the internal control nucleic acid and/ or of a member of the group of target nucleic acids as a function of the temperature of the respective sample whereby the fluorescence signal specific for the internal control nucleic acid is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the first pair of probes and the first acceptor fluorescent label of the second probe of the first pair of probes, and - whereby the fluorescence specific for a member of the group of target nucleic acids is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the member of the group of pairs of probes and the first acceptor fluorescent label of the second probe of the member of the group of pairs of probes.
- a group of pairs of probes each member hybridizing to a member of the group of target nucleic acids wherein the members of the group of pairs of probes hybridize to the respective member of the group of target nucleic acids within no more than five nucleotides distance to one another, wherein the first probe of a member of the group of pairs of probes is labeled with a second donor fluorescent label and wherein the second probe of a member of the group of pairs of probes is labeled with a corresponding second acceptor fluorescent label and wherein the hybridization of the first and second probe to the member of the group of target nucleic acids brings them into in a resonance energy transfer relationship; and - a thermostable nucleic acid polymerase and reagents necessary for amplifying the internal control nucleic acid and the group of target nucleic acids, d2) amplifying in the samples the internal control nucleic acid and the group of target nucleic acids if present in the respective sample, d3) determining in each sample separately the presence or absence of a fluorescence signal of the internal control nucleic acid and/ or of a member of the group of target nucleic acids as a function of the temperature of the respective sample whereby the fluorescence signal specific for the internal control nucleic acid is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the first pair of probes and the first acceptor fluorescent label of the second probe of the first pair of probes, and - whereby the fluorescence specific for a member of the group of target nucleic acids is generated by fluorescence resonance energy transfer between the first donor fluorescent label of said first probe of the member of the group of pairs of probes and the first acceptor fluorescent label of the second probe of the member of the group of pairs of probes.
32. The method according to any of the claims 25 to 31 wherein the label is a fluorescein dye, a rhodamine dye, a cyanine dye, or a coumarin dye..
33. The method according to any of the claims 25 to 32 wherein a member of the group of target nucleic acids comprises a nucleic acid sequence specific for a microorganism, a cell or a virus.
34. The method according to claim 33 wherein the microorganism is a gram-positive or gram-negative microorganism or a fungi.
35. The method according to claim 34 wherein a) the gram-positive microorganism is Proteus mirabilis, Serratia marcescens, Acinetobacter baumannii, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Pseudomonas aeruginosa, or Stenotrophomonas maltophilia, b) the gram-negative microorganism is a Staphylococcus spp., Enterococcus faecium or faecalis or a Streptococcus spp., or c) the fungi is Candida albicans, Aspergillus fumigatus, Candida krusei, Candida glabrata, Candida parapsilosis, or Candida tropicalis.
36. A method for verifying the determination of a signal indicating the presence of a member of a group of target nucleic acids comprising the steps of a) providing - a sample suspected to comprise a member of the group of target nucleic acids and comprising an internal control nucleic acid, - a reagent control sample comprising the group of target nucleic acids and optionally an internal control nucleic acid, - a negative control sample not comprising a member of the group of target nucleic acids and comprising an internal control nucleic acid, and - a positive control sample comprising a member of the group of target nucleic acids and an internal control nucleic acid, b) determining in each sample the signal of the internal control nucleic acid and of a member of the group of target nucleic acids, c) verifying the presence of the signal of a member of the group of target nucleic acids in the sample suspected to comprise a member of the group of target nucleic acids by:
- checking the sample suspected to comprise a member of the group of target nucleic acids for the presence of a signal of a member of the target nucleic acids independently from the presence of a signal of the internal control nucleic acid or checking the sample for the presence of a signal of the internal control nucleic acid in the case of an absence of a signal of a member of the group of target nucleic acids, - checking the reagent control sample for the presence of a signal of each member of the group of target nucleic acids, - checking the negative control sample for the absence of a signal of a member of the group of target nucleic acids and for the presence of a signal of the internal control nucleic acid, and checking the positive control sample for the presence of a signal of the target biomolecule and for the presence of a signal of the internal control nucleic acid.
- checking the sample suspected to comprise a member of the group of target nucleic acids for the presence of a signal of a member of the target nucleic acids independently from the presence of a signal of the internal control nucleic acid or checking the sample for the presence of a signal of the internal control nucleic acid in the case of an absence of a signal of a member of the group of target nucleic acids, - checking the reagent control sample for the presence of a signal of each member of the group of target nucleic acids, - checking the negative control sample for the absence of a signal of a member of the group of target nucleic acids and for the presence of a signal of the internal control nucleic acid, and checking the positive control sample for the presence of a signal of the target biomolecule and for the presence of a signal of the internal control nucleic acid.
37. The method according to claim 36 - wherein the internal control nucleic acid comprises a part of a member of the group of target nucleic acids, - wherein the positive control sample comprises a virus, a microorganism or a cell containing the target nucleic acid or wherein the positive control sample is a solution comprising a member of the group of target nucleic acids or a part thereof, or - wherein the reagent control sample is a solution comprising a member of the group of target nucleic acids or a part thereof.
38. The method according to any of the claims 36 to 37 wherein the target nucleic acid and the internal control nucleic acid is amplified before step c).
39. The method according to any of the claims 36 to 38 wherein the signal of a member of the group of target nucleic acids or of the internal control nucleic acid is a fluorescence signal.
40. The method according to claim 39 wherein the fluorescence signal is generated by a label attached to a probe that hybridizes to a member of the group of target nucleic acids or the internal control nucleic acid.
41. Use of a reagent control sample optionally comprising an internal control biomolecule and a positive control sample comprising an internal control biomolecule, both samples comprising the target biomolecule, for detecting the presence or the absence of a target biomolecule in a sample suspected to comprise the target biomolecule or for verifying the determination of a signal indicating the presence of a target biomolecule.
42. A kit for the detection of a target nucleic acid or a member of a group of target nucleic acids comprising a) a reagent control sample comprising the target nucleic acid or the member of a group of target nucleic acids, b) a negative control sample not comprising the target nucleic acid or the member of a group of target nucleic acids, c) a positive control sample comprising the target nuclecic acid or the member of the group of target nucleic acids, d) an internal control nucleic acid, and e) reagents for detecting the target nucleic acid or the member of a group of target nucleic acids.
43. The kit according to claim 42 wherein the reagents for detecting the target nucleic acid or the member of the group of target nucleic acids comprise - a pair of probes and a pair of primers, - a thermostable nucleic acid polymerase, and - reagents for amplifying the target nucleic acid or the member of a group of target nucleic acids.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP05028004.9 | 2005-12-21 | ||
| EP05028004 | 2005-12-21 | ||
| PCT/EP2006/012215 WO2007071365A1 (en) | 2005-12-21 | 2006-12-19 | Controls for nucleic acid testing |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2631425A1 true CA2631425A1 (en) | 2007-06-28 |
| CA2631425C CA2631425C (en) | 2012-10-23 |
Family
ID=36360656
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2631425A Active CA2631425C (en) | 2005-12-21 | 2006-12-19 | Controls for nucleic acid testing |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US7981606B2 (en) |
| EP (1) | EP1966395B1 (en) |
| JP (1) | JP5099920B2 (en) |
| CN (1) | CN101331237B (en) |
| CA (1) | CA2631425C (en) |
| ES (1) | ES2549410T3 (en) |
| HK (1) | HK1124888A1 (en) |
| WO (1) | WO2007071365A1 (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010071147A1 (en) * | 2008-12-16 | 2010-06-24 | アークレイ株式会社 | Method for detecting controls for nucleic acid amplification, and use thereof |
| CA2802548C (en) * | 2010-07-29 | 2017-01-03 | F. Hoffmann-La Roche Ag | Qualitative and quantitative detection of microbial nucleic acids |
| EP2465945A1 (en) * | 2010-12-17 | 2012-06-20 | F. Hoffmann-La Roche AG | Generic matrix for control nucleic acids |
| EP2554551A1 (en) * | 2011-08-03 | 2013-02-06 | Fundacio Institut mar d'Investigacions Médiques (IMIM) | Mutations in the epidermal growth factor receptor gene |
| EP2660332B1 (en) * | 2012-05-01 | 2017-02-22 | F. Hoffmann-La Roche AG | Positive control concept |
| EP3415638A1 (en) | 2012-10-18 | 2018-12-19 | IDEXX Laboratories, Inc. | Nucleic acid amplification controls and kits and methods of use thereof |
| WO2014152937A1 (en) | 2013-03-14 | 2014-09-25 | Ibis Biosciences, Inc. | Nucleic acid control panels |
| GB2525024A (en) * | 2014-04-10 | 2015-10-14 | Vela Operations Pte Ltd | Universal controls for sequencing assays |
| EP3208333B1 (en) * | 2014-10-17 | 2020-12-16 | National University Corporation Tokyo Medical and Dental University | Method for detecting and quantifying target nucleic acid in test sample using novel positive control nucleic acid |
| WO2016153355A2 (en) * | 2015-03-26 | 2016-09-29 | Microbiome Limited | Novel isolation and amplification process control |
| CN112567245A (en) * | 2018-08-28 | 2021-03-26 | Jl美迪乐博斯公司 | Method and kit for detecting target substance |
| CN114214395A (en) * | 2021-12-31 | 2022-03-22 | 上海星耀医学科技发展有限公司 | Nucleic acid amplification method and kit for improving gene detection accuracy |
Family Cites Families (50)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4458066A (en) * | 1980-02-29 | 1984-07-03 | University Patents, Inc. | Process for preparing polynucleotides |
| US4786600A (en) * | 1984-05-25 | 1988-11-22 | The Trustees Of Columbia University In The City Of New York | Autocatalytic replication of recombinant RNA |
| ES8706823A1 (en) | 1985-03-28 | 1987-06-16 | Cetus Corp | Process for amplifying, detecting, and/or cloning nucleic acid sequences. |
| US4683202A (en) * | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
| US4876187A (en) * | 1985-12-05 | 1989-10-24 | Meiogenics, Inc. | Nucleic acid compositions with scissile linkage useful for detecting nucleic acid sequences |
| US5011769A (en) * | 1985-12-05 | 1991-04-30 | Meiogenics U.S. Limited Partnership | Methods for detecting nucleic acid sequences |
| US4996143A (en) * | 1985-12-23 | 1991-02-26 | Syngene, Inc. | Fluorescent stokes shift probes for polynucleotide hybridization |
| US5403711A (en) * | 1987-11-30 | 1995-04-04 | University Of Iowa Research Foundation | Nucleic acid hybridization and amplification method for detection of specific sequences in which a complementary labeled nucleic acid probe is cleaved |
| AU622426B2 (en) | 1987-12-11 | 1992-04-09 | Abbott Laboratories | Assay using template-dependent nucleic acid probe reorganization |
| CA1340807C (en) | 1988-02-24 | 1999-11-02 | Lawrence T. Malek | Nucleic acid amplification process |
| WO1989009835A1 (en) | 1988-04-08 | 1989-10-19 | The Salk Institute For Biological Studies | Ligase-based amplification method |
| ATE144556T1 (en) | 1988-06-24 | 1996-11-15 | Amgen Inc | METHOD AND MEANS FOR DETECTING NUCLEIC ACID SEQUENCES |
| US5130238A (en) * | 1988-06-24 | 1992-07-14 | Cangene Corporation | Enhanced nucleic acid amplification process |
| JP2955759B2 (en) | 1988-07-20 | 1999-10-04 | セゲブ・ダイアグノスティックス・インコーポレイテッド | Methods for amplifying and detecting nucleic acid sequences |
| US5185243A (en) * | 1988-08-25 | 1993-02-09 | Syntex (U.S.A.) Inc. | Method for detection of specific nucleic acid sequences |
| US5118801A (en) * | 1988-09-30 | 1992-06-02 | The Public Health Research Institute | Nucleic acid process containing improved molecular switch |
| NL8900725A (en) | 1989-03-23 | 1990-10-16 | Az Univ Amsterdam | METHOD AND COMBINATION OF AGENTS FOR INSULATING NUCLEIC ACID. |
| CA2020958C (en) * | 1989-07-11 | 2005-01-11 | Daniel L. Kacian | Nucleic acid sequence amplification methods |
| US5573907A (en) * | 1990-01-26 | 1996-11-12 | Abbott Laboratories | Detecting and amplifying target nucleic acids using exonucleolytic activity |
| DE69118930T2 (en) | 1990-01-26 | 1997-01-09 | Abbott Lab | Improved method for the amplification of nucleic acid target sequence, usable for the polymerase and ligase chain reaction |
| US5210015A (en) * | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
| EP0557456A4 (en) | 1990-11-14 | 1995-11-15 | Siska Diagnostics Inc | Non-isotopic detection of nucleic acids using a polystyrene support-based sandwich hybridization assay and compositions useful therefor |
| US5455166A (en) * | 1991-01-31 | 1995-10-03 | Becton, Dickinson And Company | Strand displacement amplification |
| US5846717A (en) * | 1996-01-24 | 1998-12-08 | Third Wave Technologies, Inc. | Detection of nucleic acid sequences by invader-directed cleavage |
| DE69231853T2 (en) * | 1991-11-07 | 2001-09-13 | Nanotronics Inc | HYBRIDIZING POLYNUCLEOTIDS CONJUGED WITH CHROMOPHORES AND FLUOROPHORS TO GENERATE A DONOR-TO-DONOR ENERGY TRANSFER SYSTEM |
| US5614402A (en) * | 1992-12-07 | 1997-03-25 | Third Wave Technologies, Inc. | 5' nucleases derived from thermostable DNA polymerase |
| US5541311A (en) * | 1992-12-07 | 1996-07-30 | Third Wave Technologies, Inc. | Nucleic acid encoding synthesis-deficient thermostable DNA polymerase |
| US5719028A (en) * | 1992-12-07 | 1998-02-17 | Third Wave Technologies Inc. | Cleavase fragment length polymorphism |
| AU7487294A (en) | 1993-08-18 | 1995-03-14 | Id Biomedical Corporation | Compositions and methods for detecting target nucleic acid sequences utilizing flanking sequence enzyme molecules |
| WO1995014106A2 (en) | 1993-11-17 | 1995-05-26 | Id Biomedical Corporation | Cycling probe cleavage detection of nucleic acid sequences |
| JP3175110B2 (en) * | 1994-02-07 | 2001-06-11 | オーキッド・バイオサイエンシーズ・インコーポレイテッド | Genetic bit analysis of ligase / polymerase mediated single nucleotide polymorphisms and their use in genetic analysis |
| US5710029A (en) * | 1995-06-07 | 1998-01-20 | Gen-Probe Incorporated | Methods for determining pre-amplification levels of a nucleic acid target sequence from post-amplification levels of product |
| US5705365A (en) * | 1995-06-07 | 1998-01-06 | Gen-Probe Incorporated | Kits for determining pre-amplification levels of a nucleic acid target sequence from post-amplification levels of product |
| FR2740781B1 (en) * | 1995-11-06 | 1998-03-20 | Aremas Ass Pour La Rech L Etud | METHOD AND KIT FOR QUANTIFICATION AND DETECTION OF MICROORGANISMS |
| EP1493826B1 (en) | 1996-06-04 | 2008-03-26 | University Of Utah Research Foundation | Container for carrying out and monitoring biological processes |
| ATE318327T1 (en) | 1996-06-04 | 2006-03-15 | Univ Utah Res Found | FLUORESCENCE-DONOR-ACCEPTOR PAIR |
| US5994078A (en) | 1997-07-31 | 1999-11-30 | Maine Medical Center | Stable encapsulated reference nucleic acid and method of making |
| DE19743518A1 (en) | 1997-10-01 | 1999-04-15 | Roche Diagnostics Gmbh | Automated, universally applicable sample preparation method |
| US6395470B2 (en) * | 1997-10-31 | 2002-05-28 | Cenetron Diagnostics, Llc | Method for monitoring nucleic acid assays using synthetic internal controls with reversed nucleotide sequences |
| US20010016317A1 (en) * | 1999-09-13 | 2001-08-23 | Dolores M. Berger | Method for providing long term stability to cells for diagnostic testing |
| WO2001037291A1 (en) | 1999-11-17 | 2001-05-25 | Roche Diagnostics Gmbh | Magnetic glass particles, method for their preparation and uses thereof |
| GB0021303D0 (en) | 2000-08-30 | 2000-10-18 | Medinnova Sf | Use |
| JP3752488B2 (en) | 2000-09-22 | 2006-03-08 | 株式会社日立製作所 | Nucleic acid analysis method |
| EP1236805B1 (en) | 2001-03-02 | 2008-05-07 | Roche Diagnostics GmbH | A method for the determination of a nucleic acid using a control |
| AU2002367465A1 (en) * | 2001-07-09 | 2003-09-29 | Children's Hospital Medical Center Of Akron | Multiple controls for molecular genetic analyses |
| EP1579002B1 (en) | 2002-12-13 | 2014-02-12 | Geneohm Sciences Canada, Inc. | Methods to verify the efficiency of sample preparation and nucleic acid amplification and/or detection |
| JP4805158B2 (en) | 2003-05-16 | 2011-11-02 | アメリカ合衆国 | Internal control nucleic acid molecules for use in nucleic acid amplification systems |
| US20050029031A1 (en) | 2003-08-04 | 2005-02-10 | Thomas Ralph M. | Vehicle |
| ES2332712T3 (en) | 2003-12-16 | 2010-02-11 | Bio-Rad Pasteur | OLIGONUCLEOTIDES FOR THE DETECTION OF THE HEPATITIS B VIRUS. |
| GB0409771D0 (en) * | 2004-04-30 | 2004-06-09 | Mabtech Ab | Assay |
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