Seegene, Inc.

Chun, Jong Yoon

Lee, Young Jo

Jong Yoon Chun

Young Jo Lee

In Taek Hwang

Han Bit Lee

Seung Jae Lee

Sang Kil Lee

Mi Hyun Jang

Won Jun Oh

Gi-Seok Yoon

Yong Jo Lee

The present invention relates to a target discriminative probe (TD probe) and its uses or applications. The TD probe is hybridized with a target nucleic acid sequence through both of the 5'-second hybridization portion and the 3′-first hybridization portion. When the TD probe is hybridized with a non-target nucleic acid sequence, both the 5′-second hybridization portion and the separation portion are not hybridized with the non-target nucleic acid sequence such that both portions form a single strand due to its low Tm value. As such, the TD probe exhibits distinctly different hybridization patterns for each of the target and the non-target nucleic acid sequence, discriminating the target nucleic acid sequence from the non-target nucleic acid sequence with much higher specificity.

TD probe and its uses

The present invention relates to detection of target nucleic acid sequences using different detection temperatures. The present invention employing different detection temperatures enables to detect a plurality of target nucleic acid sequences in conventional real-time manners even with a single type of label in a single reaction vessel. The conventional technologies detect a plurality of target nucleic acid sequences by a melting analysis after target amplification. Unlikely, the present invention does not require a melting analysis after target amplification, such that the time for analysis is greatly reduced.

Detection of target nucleic acid sequences using different detection temperatures

The present invention relates to a method for calibrating a data set of a target analyte in a sample, wherein a normalization coefficient for calibrating the data set is provided by using a reference value, a reference cycle and the data set, and the calibrated data set is obtained by applying the normalization coefficient to the signal values of the data set. The present method is very effective in removing the inter- and intra-instrument signal variations of data sets. Furthermore, since the present method can be configured in software, the instant method is capable of being applied universally to various analytical instruments (e.g., a real-time PCR instrument) regardless of manufacturer. Accordingly, the method by the present invention would be very useful in diagnostic data analysis.

Method for calibrating a data set of a target analyte using a normalization coefficient

The present invention relates to differentiating signals of interest for target nucleic acid sequences. The present invention permits to obtain an individual signal value (i.e., variable) contained in a total signal detected at detection temperatures by using mathematical equations. The present invention based on equation-solving approach enables to obtain the individual signal value in a systematical manner, thereby providing analysis results in much more accurate and convenient manner.

Differentiation of signals for target nucleic acid sequences

The present invention relates to a method for de-tecting an abnormal signal using two or more datasets. The present invention makes it possible to detect abnormal signals based on characteristics of abnormal signals commonly occurring in two or more datasets, which is effectively applied to datasets obtained by a multiplex PCR method.

Detection of abnormal signal using two or more datasets

The present invention relates to the detection of a nucleotide variation on a target nucleic acid sequence using an amplification blocker and a VD-PTOCE (Variation Detection by PTO Cleavage and Extension) assay. The present invention is significantly effective in the detection of a minority mutation in an excess of wild-type DNA.

Detection of nucleotide variation on target nucleic acid sequence

The present invention relates to an apparatus for performing a nucleic acid amplification reaction and a fluorescence detection device for reaction analysis. The nucleic acid amplification apparatus of the present invention uses a plurality of blocks having different reaction temperatures by independent temperature control and the movement between the blocks is performed along sliding recesses formed in the blocks, enabling to greatly shorten the total amplification time (TAT). In the fluorescence detection device of the present invention, the positions of the light source and the photodetector are very unique for the reaction vessel in which an excitation light is provided and an emission light is generated.

Apparatus for amplificating nucleic acid and fluorescence-detecting device

The present invention relates to extraction of a signal for a target nucleic acid sequence from signals for two target nucleic acid sequences in a sample. The present invention can contribute to dramatic improvement in methods for detecting target nucleic acid sequences using different detection temperatures and reference values. The present invention using an amended reference value as well as an initial reference value can lead to increasing the detection accuracy in methods for detecting target nucleic acid sequences using different detection temperatures and reference values.

Signal extraction for a target nucleic acid sequence

The present invention relates to the detection of a target nucleic acid sequence from a DNA or a mixture of nucleic acids by a PCE-hCTO (PTO Cleavage and Extension using hCTO) assay on a solid phase. According to the present invention, the extended duplex is formed in a liquid phase in a target-dependent manner and then its presence is detected on a solid phase. Since hCTO is not immobilized onto a solid phase, the extended duplex is more effectively formed in a liquid phase.

Detection of target nucleic acid sequence on solid phase by PTO cleavage and extension using HCTO assay

The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tvalue, which are well adoptable for detection of the presence of the target nucleic acid sequence.

Detection of target nucleic acid sequences by PTO cleavage and extension assay

The present invention relates to the detection of a target nucleic acid sequence by a PCEC (PTO Cleavage and Extension-Dependent Cleavage) assay. The present invention is characterized by generating a cleavage site for a nucleolytic enzyme on the extended duplex of which the formation is dependent on the presence of a target nucleic acid sequence. The present invention detects the occurrence of the cleavage of the extended duplex, thereby determining the presence of the target nucleic acid sequence.

Detection of target nucleic acid sequence by PTO cleavage and extension-dependent cleavage

The present invention relates to technologies for preparing a tagging oligonucleotide. By analyzing exquisitely a non-complementarity level of a first tagging part, the first aspect of the present invention permits to more efficiently and easily select a suitable tagging sequence among a multitude of sequences generated theoretically. In addition, according to the second aspect of the present invention, when a nucleotide sequence for a tagging portion is first selected, one or more regions in a target nucleic acid sequence having a non-complementarity level to the nucleotide sequence for the tagging portion are found, and then a nucleotide sequence for a targeting portion is determined, tagging oligonucleotides for detecting various target nucleic acid sequences can prepared by using the fewest number of nucleotide sequences for the tagging portion and a third template.

Methods for preparing tagging oligonucleotides

The present invention relates to a method for reducing a noise level of a data set for a target analyte in a sample. The present invention can reduce a noise level of a data set to a proper level conveniently by applying a noise-reduction ratio to the data set thereby the possibility of false positive may be reduced effectively. According to the present invention, the calibrated data set is obtained by using the noise-reduction ratio, so that the noise level of a data set is reduced without change of signal ratio between the data point in the data set.

Method for reducing noise level of data set for a target analyte

The present invention relates to various processes by a template-dependent extension reaction using a dual specificity oligonucleotide and a dual specificity oligonucleotide composed of three different Tm portions therefor. Demonstrated in the present invention are the features of the dual specificity oligonucleotide, which are high hybridization specificity and mismatch tolerance.

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Seoul, South Korea

Jong Yoon Chun