Automated Brightfield Dual-Color In Situ Hybridization for Detection of Mouse Double Minute 2 Gene Amplification in Sarcomas

Abstract

These results show that the use of the ISH MDM2 and CHR12 assay allows simultaneous analyses of the 2 DNA targets within the context of tissue morphology. This method combines the advantages of a fully automated protocol with bright field microscopy and has the potential for routine application in surgical pathology.

Comparison of Chromogenic In Situ Hybridization and Fluorescence In Situ Hybridization for the Evaluation of MDM2 Amplification in Adipocytic Tumors

“…This compares to Zhang et al, who reported a failure of CISH in two out of 100 cases and attributed this failure to tissue inadequacy (6). In both of our cases, both black and red (control and locus-specific) probes were weak or undetectable, in both the tumor cells and surrounding non-neoplastic stromal cells.…”

“…The assay performance was evaluated on a cohort of 100 formalin-fixed, paraffin-embedded soft tissue specimens, and excellent sensitivity and specificity were reported (6). Thus far, a direct comparison has not been made between FISH and CISH results in the same tumor samples.…”

Mouse double minute 2 amplification in oesophageal squamous cell carcinoma is associated with better outcome

“…The average number of MDM2 and chromosome 12 centromere‐specific ( CEP12 ) signals was then determined, and the MDM2/CEP12 ratio was calculated for each case. Using the manufacturer's instructions for dual‐colour in‐situ hybridisation and previous findings on soft‐tissue sarcomas, small clusters of multiple signals were counted as six signals, large clusters were counted as 12 signals and a ratio greater than 2.0 was considered to indicate MDM2 amplification 23,24 . A ratio less than 2.0 was considered to indicate non‐amplification, and a ratio less than 2.0 with ≥2 signals of both probes was considered to indicate polysomy.…”

Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Lateral-Flow Immunochromatographic Strip for the Detection of Digoxin in Human Blood

“…The enzyme-linked immunosorbent assay (ELISA) provides the ability to process huge numbers of samples with high sensitivity and specificity and convenience. − The most effective way to prevent DIG poisoning is to apply an anti-DIG antibody. , In recent years, anti-DIG antibodies also play an important role in molecular biology hybridization techniques and can be used to detect all nucleic acids, proteins, and carbohydrates labeled by DIG, for example, Teles et al developed an in situ hybridization assay to detect the MDM2 gene amplification by using a dinitrophenyl-labeled MDM2 probe and a DIG-labeled CHR12 probe on an automated slide staining platform at Ventana Medical Systems. , Although polyclonal antibodies are sensitive and specific, they are not widely used in the market because they have poor reproducibility. Furthermore, anti-DIG monoclonal antibodies (mAbs) have been shown to be more useful than polyclonal antisera in the clinical management of patients with heart disease and in reversing the toxicity of digitalis.…”