Burkhardt Anne, Smith Chelsey L, Singh Rajesh R
Department of Molecular Oncology, Quest Diagnostics Nichols Institute, Chantilly, VA 20151, USA.
Diagnostics (Basel). 2025 Feb 6;15(3):390. doi: 10.3390/diagnostics15030390.
Accurate screening of clinically significant tumor mutations is critical for precision medicine in oncology. This requires genotyping platforms with high accuracy and compatibility with varying DNA yields from challenging sample types. Here, we have validated a new, improved, compact, and self-contained pyrosequencing platform (Pyromark Q48 Autoprep; Q48) for screening -, - and mutations in thyroid cancers. A set of 73 thyroid cancer and 16 non-thyroid cancer samples (fine needle aspirates and formalin-fixed paraffin-embedded) with known mutation status of genes were tested using the Q48 platform. Performance parameters such as accuracy, precision, and limit-of-detection were established. Q48 workflow was compared to an older Q96 pyrosequencing platform to highlight the differences and advantages. testing by pyrosequencing was also compared to a clinically validated next-generation sequencing platform using 56 thyroid cancer samples. The Q48 Pyromark was found to be a very reliable platform suited for quick testing of genes with complete accuracy, high precision, and high detection sensitivity. It had comparable accuracy, with higher sequencing success rates than NGS. The hands-on time, workflow ease, and efficiency were also significantly improved in comparison with the Q96 platform. Overall, the Q48 platform was found to be a well-suited and agile clinical sequencing platform to rapidly screen mutations.
准确筛查具有临床意义的肿瘤突变对于肿瘤学的精准医疗至关重要。这需要具有高精度且能与来自具有挑战性样本类型的不同DNA产量相兼容的基因分型平台。在此，我们已验证了一种新型、改进、紧凑且自成一体的焦磷酸测序平台（Pyromark Q48 Autoprep；Q48），用于筛查甲状腺癌中的 、 和 突变。使用Q48平台对一组73份已知 基因突变状态的甲状腺癌样本和16份非甲状腺癌样本（细针穿刺抽吸物和福尔马林固定石蜡包埋样本）进行了检测。确定了诸如准确性、精密度和检测限等性能参数。将Q48工作流程与较旧的Q96焦磷酸测序平台进行比较，以突出差异和优势。还使用56份甲状腺癌样本将焦磷酸测序检测与经过临床验证的下一代测序平台进行了比较。发现Q48 Pyromark是一个非常可靠的平台，适合对 基因进行快速检测，具有完全的准确性、高精度和高检测灵敏度。其准确性相当，测序成功率高于下一代测序。与Q96平台相比，实际操作时间、工作流程的简便性和效率也有显著提高。总体而言，发现Q48平台是一个非常适合且灵活的临床测序平台，可快速筛查 突变。
