NuProbe Technology Showcases Quantitative PCR for Simultaneous Enrichment and Identification of Multiple Rare Variants below 0.1% VAF

January 18, 2022

Houston, Jan. 18, 2022 –NuProbe, a genomics and molecular diagnostics company developing ultrasensitive assays for precision oncology research, published research demonstrating new technology to qualitatively and quantitively detect multiple low-frequency variants in one quantitative PCR (qPCR) reaction. The new allele-specific BDA technology (As-BDA) uses qPCR to detect mutations down to 0.01% variant allele fraction (VAF) within 2 hrs.

Blocker Displacement Amplification (BDA), one of NuProbe’s core technologies, removes wildtype sequences from the library and selectively enriches rare variants, achieving a limit of detection (LoD) of 0.1% VAF in multiplexed real-time PCR and NGS. With standard BDA and real-time PCR, mutations within the blocker enrichment region can be identified with more reactions or another sequencing technology such as Sanger sequencing. As-BDA improves upon BDA by precisely reporting the specific mutations within a sample at an improved limit of detection.

Read more about BDA.

“As-BDA overcomes the limitations of current technologies, such as NGS and droplet digital PCR, in terms of sensitivity, multiplexity, and workflow simplicity,” says Kerou Zhang, a consulting scientist of NuProbe and the first author of the research. “As-BDA identifies variants with high sensitivity and specificity, making it advantageous and potential in non-invasive mutation profiling with liquid biopsy for cancer diagnosis and monitoring, where mutation VAF is usually very low.”

Studies were done on a three-tube IDH2 assay and a single-tube analysis of IDH2 R140Q. For the three-tube assay, each tube detected up to 4 mutations simultaneously with as low as 0.1% VAF using 15ng DNA input. The scientists also completed comparison studies with ddPCR for R140Q mutation using peripheral blood mononuclear cell (PMBC) DNA from 11 acute myeloid leukemia (AML) clinical samples, 7 healthy donor samples, and reference samples. As-BDA results were 100% concordant with ddPCR for mutations at as low as 0.1% VAF.

“We are excited about this new technology that can provide concurrent qualitative and quantitative mutation detection with a rapid turnaround time,” says David Zhang, General Manager of NuProbe and the corresponding author of the research. “When translated into molecular diagnostics and genomics research, this technology has the potential to improve clinical sensitivity with rapid turnaround time.”


About NuProbe

NuProbe is a cutting-edge genomics and molecular diagnostics company with revolutionary molecular diagnostic technologies to improve the sensitivity of sequencing mutations and copy number variations by over 10-fold. NuProbe has sites in Houston, USA, Shanghai, China and Suzhou, China. NuProbe’s vision is to offer affordable, timely, and accurate disease state information to enable precision medicine and improve patient outcomes.

NuProbe’s products are for Research Use Only and are not intended for In Vitro Diagnostic use.

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