Real-time reverse transcription PCR and the detection of occult disease in colorectal cancer

RNA biomarkers in colorectal cancer

“…Local extent of disease and 54 metastasis to regional lymph nodes (LNs) constitute the basis for 55 histopathological staging, which takes into account different de- 56 grees of penetration of the primary tumour through the bowel wall 57 (T), the degree of LN involvement (N) and the absence or presence 58 of distant metastasis (M) [2]. The American Joint Committee on 59 Cancer (AJCC) has condensed the TNM system into four stages, 60 ranging from stage I defined as early cancer through to advanced 61 metastatic stage IV disease (Fig. 1).…”

Detection of Genetic Alterations by Nucleic Acid Analysis: Use of PCR and Mass Spectroscopy-Based Methods

“…By preparing all nucleic acid samples in a highly defined and careful manner, the enzymatic reactions on which PCR, qPCR, RT qPCR depend are allowed to occur as intended [3]. A better knowledge of the DNA template structure and composition is also a key for getting optimal and accurate results in qPCR.…”

Molecular Medicine, Gene-Expression Profiling and Molecular Diagnostics: Putting the Cart Before the Horse

“…Another important factor in the handicapping of molecular diagnostics utilizing mRNAexpression profiling data is the lack of established quality control networks [45]. Quality assessment issues range from appropriate sample selection and handling, through RNA quality control, cDNA synthesis protocols, data handling and interpretation all the way to monitoring of the quality of the laboratories carrying out the molecular assays [46]. I consider there to be two sets of confounding parameters, broadly falling into biological and technical categories, which combine to make it difficult to characterize accurately or comprehensively a tumor from a single, small tissue biopsy.…”

“…Leaving aside the question of tumor specificity of the detected mRNA, without knowledge of the metastatic potential such information merely identifies the tumor burden, which may or may not be related to the likelihood of metastasis development [46].…”