APPLICATION NOTE qPCR MIXES FOR tNGS
STUDY RESULTS
The study assessed two PCR chemistries in a hereditary breast cancer assay to determine performance across critical sequencing metrics. Using a two-step, 177-target multiplex qPCR approach, BRCA1 and BRCA2 regions were amplified with Meridian’s Lyo-Ready Direct DNA qPCR Blood Mix [MDX122] and a commercial alternative. After library preparation, sequencing on the DNBSEQ-G400 platform showed that both mixes performed strongly, with mapping rates exceeding 99% and coverage uniformity above 85%. These results confirm that Meridian’s optimized chemistry enables reliable amplification and well-balanced libraries—essential for accurate variant detection and consistent performance in tNGS workflows.
The results showed equivalent performance across all critical metrics, confirming that a mix validated for qPCR can also power complex tNGS workflows . For developers, this demonstrates the versatility of Meridian’s chemistry: one mix, multiple applications—helping simplify validation, reduce risk and streamline assay development pipelines. Meridian’s Lyo-Ready Direct DNA qPCR Blood Mix delivered uniform amplification and supported accurate detection of BRCA1 and BRCA2 variants— even at low frequencies from low-input DNA .
KEY SEQUENCING OUTCOMES
Meridian’s Mix Commercial Mix
MAPPING RATE
99.9% 99.9% 99.9% 99.6%
TARGET RATE
UNIFORMITY
86.7% 88.6%
0%
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80%
100%
TARGETED NGS (tNGS) A GROWING FORCE IN PRECISION DIAGNOSTICS Targeted next-generation sequencing (tNGS) is rapidly becoming a cornerstone of precision medicine, bridging the gap between single-target assays and genome-wide approaches. It offers high multiplexing while maintaining
manageable data complexity. Clinical practice now relies heavily on tNGS, with many FDA- approved commercial assays for solid tumors, hematologic malignancies and liquid biopsy applications widely adopted in both centralized and specialty oncology labs. 3 This growing clinical use is fueling significant market expansion—the global NGS market is projected to rise from $12.9 billion in 2023 to nearly $98 billion by 2035. 4
With FDA-approved tests already in use and a market projected to hit $98B by 2035 , tNGS is transforming precision diagnostics .
EXAMPLES OF FDA-CLEARED TARGETED NGS ASSAYS USED IN CLINICAL PRACTICE
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