Sadaqat Zoha, Joseph Smitha, Verma Chandrika, Muni Reddy Jyothi, Prakash Anand, Thomas Tinku, Bharadwaj Vandana, Vyas Neha
Manipal Academy of Higher Education (MAHE), Manipal 576104, Karnataka, India.
Division of Molecular Medicine, St John's Research Institute, St John's National Academy of Health Sciences (a Unit of CBCI Society for Medical Education), Bangalore 560034, Karnataka, India.
Biol Methods Protoc. 2024 Dec 28;10(1):bpae098. doi: 10.1093/biomethods/bpae098. eCollection 2025.
Real time-polymerase chain reaction (RT-PCR) is used routinely in clinical practice as a cost-effective method for molecular diagnostics. Research in pediatric B-cell Acute Lymphoblastic Leukemia (ped B-ALL) suggests that apart from cytogenetics and clinical features, there is a need to include Copy number variation (CNV) in select genes at diagnosis, for upfront stratification of patients. Using ped B-ALL as a model, we have developed a RT-PCR-based iterative probability scoring method for reporting CNVs, and relative gene-expression changes. Our work highlights that once genes of interest and hotspots of CNVs are identified in discovery phase, our proposed method can be used as a cost-effective and user-friendly diagnostic tool for the identification of changes at genomic or transcriptomic level. It has the potential to be incorporated in routine diagnostics in resource constrained settings and be tailored for different diseases as per need.
实时聚合酶链反应（RT-PCR）在临床实践中被常规用作一种经济高效的分子诊断方法。儿科B细胞急性淋巴细胞白血病（ped B-ALL）的研究表明，除了细胞遗传学和临床特征外，在诊断时还需要纳入特定基因的拷贝数变异（CNV），以便对患者进行前期分层。以ped B-ALL为模型，我们开发了一种基于RT-PCR的迭代概率评分方法，用于报告CNV和相对基因表达变化。我们的工作强调，一旦在发现阶段确定了感兴趣的基因和CNV热点，我们提出的方法就可以用作一种经济高效且用户友好的诊断工具，用于识别基因组或转录组水平的变化。它有可能被纳入资源有限环境下的常规诊断，并可根据需要针对不同疾病进行定制。
