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US 2026/0098290 A1Published application
Method for detecting n target nucleic acids in sample using n detection temperatures
AssigneeSeegene, Inc.
InventorsYoung Jo LEE, Han Bit LEE, Hyun Ji HAN, Jeong Woo KIM
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Abstract
The present invention relates to a method of detecting n target nucleic acids in a sample, and more particularly, to a method of detecting n target nucleic acids in a sample in a real-time manner in one reaction vessel by being configured such that signals from one or two target nucleic acids are detected at each of n (n is an integer of 3 or more) detection temperatures despite the use of a single fluorescent label.
Claims30
A method for detecting n target nucleic acids in a sample using n detection temperatures, comprising the steps of:
- (a) reacting, in a single reaction vessel, a sample suspected of containing at least one of the n target nucleic acids with n compositions for detecting the n target nucleic acids,
- wherein n is an integer of 3 or more,
- wherein, during the reacting, the n target nucleic acids in the sample are amplified,
- wherein each of the n compositions comprises one or more oligonucleotides capable of providing a signal depending on the presence of its corresponding target nucleic acid among the n target nucleic acids, and signals provided by the n compositions are not differentiated from each other by a single detection channel,
- wherein 1 to n−1 of the n compositions are each configured to provide a signal indicative of the presence of its corresponding target nucleic acid only at one different detection temperature among the n detection temperatures in the presence of the corresponding target nucleic acid, whereas the other compositions are each configured to provide a signal indicative of the presence of its corresponding target nucleic acid at two to n adjacent detection temperatures among the n detection temperatures arranged in order in the presence of the corresponding target nucleic acid,
- wherein at each of the n detection temperatures, signals are provided by one or two of the n compositions;
- (b) measuring the signals at the n detection temperatures under the single detection channel,
- wherein none to n−1 of the n detection temperatures are single-signal detection temperatures selected such that a single signal is provided by one of the n compositions, whereas the other detection temperatures are combined-signal detection temperatures selected such that a combined signal is provided by two of the n compositions;
- and
- (c) determining the presence of a target nucleic acid from the single signal measured at the single-signal detection temperature and determining the presence of a target nucleic acid from an extracted single signal, which is obtained by extraction from the combined signal measured at the combined-signal detection temperature, whereby the presence of the n target nucleic acids are determined from the signals measured at the n detection temperatures.
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A computer readable storage medium containing instructions to configure a processor to perform a method for detecting n target nucleic acids in a sample using n detection temperatures, the method comprising:
- (a) receiving signals measured at the n detection temperatures under a single detection channel,
- wherein the signals are obtained by reacting, in a single reaction vessel, a sample suspected of containing at least one of n target nucleic acids with n compositions for detecting n target nucleic acids, and measuring the signals at the n detection temperatures under the single detection channel,
- wherein n is an integer of 3 or more,
- wherein, during the reacting, the n target nucleic acids in the sample are amplified,
- wherein each of the n compositions comprises one or more oligonucleotides capable of providing a signal depending on the presence of its corresponding target nucleic acid among the n target nucleic acids, and signals provided by the n compositions are not differentiated from each other by a single detection channel,
- wherein 1 to n−1 of the n compositions are configured to provide a signal indicative of the presence of its corresponding target nucleic acid only at one different detection temperature among the n detection temperatures in the presence of the corresponding target nucleic acid, whereas the other compositions are configured to provide a signal indicative of the presence of its corresponding target nucleic acid at two to n adjacent detection temperatures among the n detection temperatures arranged in order in the presence of the corresponding target nucleic acid,
- wherein none to n−1 of the n detection temperatures are single-signal detection temperatures selected such that a single signal is provided by one of the n compositions, whereas the other detection temperatures are combined-signal detection temperatures selected such that a combined signal is provided by two of the n compositions;
- and
- (b) determining the presence of a target nucleic acid from the single signal measured at the single-signal detection temperature and determining the presence of a target nucleic acid from a single signal, which is extracted from the combined signal measured at the combined-signal detection temperature such that the presence of the n target nucleic acids are determined from the signals measured at the n detection temperatures.
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Dependencies are inferred from the claim text.