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Seegene's Molecular Diagnostic Products are the best

Seegene’s innovative molecular diagnostic products enable to detect multiple pathogen infections simultaneously and accurately.

You can test several pathogens related to a certain diseases at the same time.
Seegene’s patented molecular diagnostic products can analyzes target gene, specific to each pathogen.

Seegene's molecular diagnostic products provide comprehensive useful information from a single test.

When you are infected with virus or bacteria, Seegene’s diagnostic products not only tell you your infection status, but also provide the quantitative (titer level) information of the pathogens in three levels - low, intermediate, high. This additional information is particularly useful for follow-up management such as forecasting or monitoring the progress of the disease or treatment.

 

Clinical applications of Seegene’s Molecular Diagnostic Products

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What are the benefits of Molecular Diagnostics?

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Molecular diagnostic is a diagnostic method that uses genetic materials to detect viral and bacterial infections. It is very effective to make the diagnosis of infectious diseases, because of fast, accurate and customized results. It also guides the best treatment method for patient management, even for complicated diagnosis such as identifying drug resistant or mutant genes.

Why seegene's Molecular Diagnostic Products are the best?

Seegene’s innovative molecular diagnostic products enable to detect multiple pathogen infections simultaneously and accurately.

You can test several pathogens related to a certain diseases at the same time.
Seegene’s patented molecular diagnostic products can analyzes target gene, specific to each pathogen.

Seegene's molecular diagnostic products provide comprehensive useful information from a single test.

When you are infected with virus or bacteria, Seegene’s diagnostic products not only tell you your infection status, but also provide the quantitative (titer level) information of the pathogens in three levels - low, intermediate, high. This additional information is particularly useful for follow-up management such as forecasting or monitoring the progress of the disease or treatment.

 

TOCE™ Technology

The New Paradigm for High multiplex Real-time PCR

High multiplex Real-time Quantitative PCR

TOCE™ technology enables confirmation of multiple (five or more) target genetic detection and genetic variation. TOCE™ can implement qualitative test that tests multiple clinical samples and quantitative analysis bases on Cyclic Catcher Melting Temperature Analysis (cyclic-CMTA).
The profile of information generated by TOCE™-based high multiplex PCR analysis can substantially accelerate informed accurate diagnosis and treatment. As such, TOCE™ -based molecular testing is causing a paradigm shift in molecular diagnostics toward the high-content/high-quality testing needed for optimized patient treatment.

 

DPO™ Technology

Novel Oligo platform of super multiplex PCR

New primer technology that inhibits combined nonspecific template.

 Proper priming between oligonucleotide primer and template is important to obtain accurate result from PCR. Dual Priming Oligonucleotide(DPO) primer inhibits occurrence of PCR product (mismatched priming, non-specific priming). A novel DPO™ system that is structurally and functionally different from the primer system currently in wide-spread use blocked extension of non-specially primed templates, and thereby generates consistently high PCR specificity even under less optimal PCR conditions. 

 

Features of the TOCE™ technology

1. Principles of TOCE™ technology

The key components for TOCE™ technology are DPO™ primer pairs, Pitchers and Catchers. The DPO™ is a Seegene's proprietary target-specific primer and provides highly specific amplification of the target region. The Pitcher is a tagging oligonucleotide which hybridizes specifically to the target region. The Catcher is a fluorescently labeled artificial template. 

Principles of TOCE™ technology

2. High multiplicity in a single channel using Catcher -Tm analysis

TOCE™ technology enables identification of multiple target analytes simultaneously in a single channel. The signal can be measured in real-time and/or analyzed by Catcher melting temperature (Catcher-Tm) to detect the presence of the target analyte. 

High multiplicity in a single channel using Catcher -Tm analysis

3. Catcher-Tm : Flexibility in the adjustment of Tm value

One unique feature of TOCE™ is the "Catcher," which is a fluorescently labeled artificial template and generates the signal for each target sequence. The Catcher-Tm value can be controlled by adjusting the sequence and length of the Catcher. For TOCE™ assay optimization, the Catcher-Tm value can be easily adjusted and is not limited by the target sequence.

Catcher-Tm : Flexibility in the adjustment of Tm value

4. Consistent Catcher-Tm value regardless of target sequence variations

Current Tm-based analysis method has different Tm value with occurrence of changes in nucleotide sequence of the pathogen. However, TOCE™ technology allows for a consistent Catcher-Tm value that is not affected by variations of the target sequence. 

Consistent Catcher-Tm value regardless of target sequence variationse

5. As sensitive as singleplex real-time PCR

The detection limits of multiple pathogens using TOCE™ technology are similar to those of probe-based singleplex real-time PCR.

Singleplex real-time PCR과 동등한 민감도* 10-plex real-time PCR : 7 analytes + dual target for CT & NG + IC (Chlamydia trachomatis; Neisseria gonorrhoeae; Trichomonas vaginalis; Mycoplasma hominis; Mycoplasma genitalium; Ureaplasma urealyticum; Ureaplasma parvum) ** Singleplex real-time PCR : 1 analyte (C. trachomatis)

6. Quantitative analysis by cyclic-CMTA

Presence of infection in infected pathogens and quantitative analyzed result can be simultaneously viewed by setting cyclic-CMTA point in the Real-time PCR process. Cyclic-CMTA point can be set at 30, 40, 50 cycle in the real-time PCR process, and quantitative analysis can be performed depending on the melting peak due to amount of infected pathogens as shown in the chart of analysis result. 

Quantitative analysis by cyclic-CMTA석Multiple quantitative analysis of HPVs can be determined by cyclic-CMTA.

The difference of appearance of the melting peak at the cyclic-CMTA point indicates different titers of target (low, intermediate, and high copy). In this example, the melting peak of HPV 33 appears at the first cyclic-CMTA point (cycle 30) corresponding to high copy number. HPV 16 appears at the second cyclic-CMTA point (cycle 40) corresponding to intermediate copy. HPV 18 appears at the third cyclic-CMTA point (cycle 50) corresponding to low copy. This measurement allows for quantitative analysis, persistence, and clearance of multiple HPVs, aiding in the diagnosis of viral infection associated with incidences of cervical cancer.

7. High multiple point mutation detection in a single tube

TOCE™ technology allows for mutation detection to discriminate even one base difference accurately, enabling detection of clustered point mutations. In this example, signal is generated ONLY when mutant-specific pitcher binds to mutated target sequence, discriminating mutations from wild-type sequence.

High multiple point mutation detection in a single tube

 

DPO™ Technology

Novel Oligo platform of super multiplex PCR

 

New primer technology that inhibits combined nonspecific template.

 

Features of the technology

Freedom in primer design & PCR optimization

DPO™ comprises of two separate priming regions (5'-end stabilizer and 3'-end determiner) joined by a polydeoxyinosine linker. The linker forms like a "bubble-like structure" which itself is not involved in priming, but rather delineates the boundary between 5'-end stabilizer and 3'-end determiner.

디자인할 수 있는 부위가 자유롭고 PCR 성능을 높여줌

Principle of DPO™
DPO™ has two functional priming regions (one is longer than the other) separated by the poly (I) linker. These two unequally distributed priming regions generate dual priming reactions based on the following scheme, resulting in only target-specific products. 


Step 1: Poly(I) linker activation
Deoxyinosine has a relatively low Tm value compared to the natural bases, due to weaker hydrogen bonding so that the poly (I) linker will form a bubble-like structure at a certain annealing temperature and separates 5'-end stabilizer and 3'-end determiner as "stabilizer" and "determiner" 

Step 2: First priming reaction
The longer 5'-segment preferentially binds to the template DNA and initiates "stable annealing". It acts as a Stabilizer".

Step 3: Second priming reaction
The short 3'-segment selectively binds to a target site and determines "target-specific extension". It acts as a "Determiner".

Comparison between DPO™ primer VS. Non-DPO™ primer

Fig. 1.Comparison of Ndufs2 products obtained using DPO™ primer and Non-DPO™ primer

Another major advantage is that DPO™ makes primer design extremely simple and easy Since two separate priming reactions provide a primer with a comfort zone (high tolerance) in annealing. Following the first stable priming reaction by Stabilizer of 5' end, the second critical priming reaction by Determiner gives one additional chance to correct the specificity. For this reason, DPO™ does not require a rigid optimization of PCR conditions and primer search parameters including primer length, GC content, annealing temperature, and secondary structure (hairpin, self or cross dimer). 

Comparison between DPO™ primer VS. Non-DPO™ primer

Unparalleled high specificity

Example 1. DPO™ specificity over a wide range of annealing temperature 

annealing 온도 변화조건에서 특이성을 보여주는 DPO™ Fig. 2. Fig 2. Non- DPO™ primer shows specificity at high specific annealing temperature, and production of false product such as non-specific band occur at low annealing temperature while the DPO™ primer produced only one target product over a change of annealing temperature. (lanes 2 and 4). 

M: Size marker
Lane 1: Non-DPO™ primer
Lane 2: DPO™
 

No primer competition and dimerization in Multiplex

DPO™ technology has high specificity without false-positive. DPO™ Multiplex system using this technology is an accurate, quick, and cost-effective molecular diagnostic method.

Multiplex-PCR 수행시 primer간의 dimer 형성 또는 competition이 없음
However, DPO™ allows specific detection of a large number of pathogens without any false result because the bubble-like structure of the poly(I) linker in DPO™ efficiently prevents primer-dimer and hairpin structure formation. DPO™-Multiplex PCR generates the high specificity without production of any non-specific bands or false-positive products and it represents a reliable, rapid, practical and cost-effective detection method



Single base discrimination

Multiple-pathogen Detection
- High specificity without production of any non-specific or false-positive results 
- Reliable, rapid, practical and cost-effective detection method
- Specific and simultaneous detection of multiple pathogens without any false results


Multiple-genotyping
- Decide genotype and subtype of various pathogens. 

Multiple-SNP Detection
- Specific and simultaneous analysis of multiple single nucleotide polymorphic (SNP) sites 

Occurrence of Single nucleotide polymorphisms (SNPs) are responsible for drug resistance leading to various diseases, but additional steps after amplification of an SNP-containing region such as RFLP, sequence or hybridization are needed to identify SNP genotyping accurately. As a result, the longer time will take for SNP genotyping and additional test will be done. High specificity of DPO enables SNP genotyping with only a single PCR by identifying SNP accurately. 

단일 염기 변이 구별 (다양한 적용 범위)

  • Hetero type of allele 1 & 2: lanes 1~3
  • Homo type of allele 1: lanes 4~6
  • Homo type of allele 2: lanes 7~9
SNP in CYP2C19 results from a single base pair substitution (681 bp:G→A) at position 681 in exon 5 of CYP2C19 and causes a non-functional protein and affects the metabolism of a number of commonly used drugs. Multiplex PCR analysis of nine human genomic DNA samples with known genotypes at the CYP2C19 locus using conventional primers (left) and DPO™ (right) was carried out. As a result, DPO™-based multiplex PCR clearly distinguished between the different alleles of CYP2C19, while conventional primer-based multiplex PCR did not.

Guaranteed reproducibility

High DPO™ specificity with GC-rich template.

GC contents가 높은 조건에서 specificity를 보이는 DPO™Fig. 1.The amplification of the human klotho target sequence containing 81 % GC was compared using DPO™ primer and Non-DPO™ primer. 

Amplification of the human GSPT1 target sequence containing 81 % GC with DPO™ primer and Non-DPO™ primer. 

A. PCR reaction excluding the detergent (5% DMSO + 1M Betaine).The conventional primers generate many non-specific products in a PCR reaction omitting the detergent, whereas the DPO™ does not amplify any PCR product. B. PCR reaction including the detergent.When the detergent are added to relax secondary structures for higher GC content, DPO™ successfully amplifies PCR product.