SuperbDNA™ Next Generation bDNA Signal Amplification Technology | DiaCarta, Inc.

SuperbDNA™ bDNA Signal Amplification Technology

A Powerful Tool for Molecular Diagnostics 

Qualitative and Quantitative DNA Copy or
Gene Expression Analysis

SuperbDNA™ technology uses branched DNA to qualitatively or quantitatively measure the presence of target DNA or RNA. It has been used in FDA-approved clinical applications including prognosis and monitoring of patients with viral diseases. Unlike PCR or RT-PCR, SuperbDNA™ technology does not amplify the DNA or RNA in the samples, rather amplify the signals for detection. Because the signal amplified is proportional to the levels of the target (DNA or RNA) in the samples, the latter is easy to be quantified with a standard curve.

Advantages of SuperbDNA™ Technology Compared with RT-PCR

Although widely used and approved as a powerful technology, PCR and RT-PCR have their drawbacks when used as molecular diagnostics tools.

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PCR and RT-PCR rely on amplification of DNA or cDNA to generate signals for diagnostic analysis. Because amplified DNA often have cross-contamination problems without proper operational control, SuperbDNA™ technology normally does not have this problem.

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PCR and RT-PCR rely on amplification of DNA or cDNA to generate signals for diagnostic analysis. Because amplified DNA often have cross-contamination problems without proper operational control, SuperbDNA™ technology normally does not have this problem.

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Both RT-PCR and branched DNA technology have been shown to be correlated well in determination of target gene expression. However, RT-PCR has greater proportional system errors and is proved to be less efficient for quantitation of some subtype of HIV virus. When the target is in low concentration, RT-PCR has better quantitation than branched DNA technology. In the meanwhile, when the target concentration increases to mid or high level, branched DNA technology shows more trustable results.

The Principle and Workflow for SuperbDNA™ or Branch DNA Technology

SuperbDNA™ technology uses a series of hybridization events to capture the target and another series of hybridization events to connect the labeled probes for chemiluminescent signal amplification. Such a “sandwich” structure efficiently captures and quantify the target in the samples with a signal amplification to measure 500 to 10 million molecules.

First, “capture probes” that are pro-coated on the bottom of plates or conjugated to beads or discs hybridize with “target probes” that binds specific target DNA or RNA sequences. Once the samples are added, the target DNA or RNA sequence will be captured and the rest of the samples are washed away. Secondly, the second set of the probes, including “preamplifier” and “amplifier” probes labeled with alkaline phosphatases covalently, are added to hybridize with the target molecules to form the “sandwich”. The second wash will wash away unhybridized probes. Finally, the substrate for alkaline phosphatase is added and chemiluminescent signals can be detected. The workflow for the detection is illustrated blow. 

Step 1: Lyse Samples

Step 2: Capture E6/E7

Step 3: Amplifier

Step 4: Label Probe - AP

Step 5: Substrates

Step 6: Read and Report

SuperbDNA™ Technology Platform: Single Assay or Multiplex Assay?

SuperbDNA™ technology can be used in a single well assay format for each target, or multiplex assays for multiple targets in the discs or beads format. Contact us for different formats available to perform your assay.

Applications for SuperbDNA™ technology

SuperbDNA™ or branched DNA technology can be used in different areas for target DNA or RNA detection directly from the samples, including pathogen detection, biomarker quantitation, gene expression, and in situ hybridization. The benefits for these assays can be summarized as below

Gene Expression Analysis

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Ideal clinical platform for companion diagnostic delivery

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Low cost of goods and rapid turn-around time

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96-well format. No RNA purification required

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Highly sensitive: detect 50 transcripts per ml

Pathogen Detection (Virus, Fungi and Bacteria)

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No requirement for target purification and amplification

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Improved sensitivity and specificity

In Situ Hybridization

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Single molecular detection

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FFPE and frozen tissue compatible

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Strong/bright signal with low background

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Multiple-4-plex easily attainable

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An ideal complement to antibody-based advance tissue staining-compatible with bright filed and fluorescent

SuperbDNA™ Featured Application: QuantiVirus™ HPV E6/E7 mRNA Test 

QuantiVirus™ HPV E6/E7 mRNA Test – Predicting Pre-Cancer in Cervical Cancer and Head & Neck Cancer Screening

Human papillomavirus (HPV), with over 40 distinct HPV types, is the most common sexually transmitted infection in the U.S. , affecting 42.5% of the adults aged 18–59 years during 2013–2014. The risk for persistent infection with some HPV types is development of cervical cancer for women and head and neck cancer in men.

 

QuantiVirus™ HPV E6/E7 mRNA test detects HPV oncogene E6/E7 mRNA from 14 high-risk types and genotyping of HPV16 and 18 directly from women cervical samples or head and neck cancer in men screening directly from saliva.

 

SuperbDNA™ Featured Application: Radtox™ cfDNA Test

RadTox™ cfDNA Test – Radiation Therapy Toxicity Monitoring

Powered by SuperbDNA™ Technology, RadTox™ cfDNA Test is developed for cancer patients’ radiation therapy toxicity monitoring, which is fulfilling an unmet medical need in cancer radiation therapy by enabling the direct monitoring of side effect severity and tumor response within days of radiation treatment initiation. RadTox™ would allow dose escalation to tumors in less sensitive patients, aid development of radiotoxicity mitigators, and reduce overtreatment risk for sensitive patients. The assay provides a fast, cost-effective quantitation of liquid biopsy cfDNA sample from cancer patients before and after radiation therapy and provides patient information for estimates of dosage usage and general radiation resistance. 

 

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