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miRNA-ISH for Cancer of Unknown Primary

miRNA-ISH for Cancer of Unknown Primary

Summary:

World-wide approximately 5% of all cancers are determined to have an occult or unknown primary location, upon initial assessment, prior to completing a full pathology report. [Oien] Unfortunately in 20-50% of CUP cases the primary tumor is never found. These cancers of unknown primary (CUP) result from metastasis. Occult cancer is most frequent in individuals around 60 years of age and an estimated 31,480 cases of CUP will be diagnosed in the US in 2019, making up around 2% of all US cancers. [Siegel] People with a family history of lung, kidney, or colorectal cancer are more likely than average to develop occult cancer. [Hemminki] Median survival time is 8-12 months, 80% of patients live only 6 months after diagnosis. [Losa]

Conclusion:

Patients with metastatic cancer are often recruited to clinical trials using investigational drugs due to a lack of effective market approved treatment options. CUPs are most often found in the liver, lung, bone, brain; lymph nodes; and peritoneal and pleural serous cavities. [Oien] Once a tumor has been discovered, in order to provide a working diagnosis for the treating oncologist, a detailed pathology report must be prepared. A crucial component of this report is a microRNA in situ hybridization (miRNA-ISH) based classification of the tumor to aide categorization and for determining the site of origin.

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MicroRNA in situ Hybridization for Cancer Diagnosis

MicroRNA in situ hybridization for cancer diagnosis

Summary: MicroRNAs (miRNA) are approximately 19–24 nucleotides in length and base-pair to complementary sites within messenger RNA (mRNA). Aided by the “RISC” complex this binding functions to promote down-regulation of the mRNA’s protein product. MicroRNAs are an important biomarker of cancer as a result of their involvement in multiple biological processes including development, differentiation, proliferation, metabolism, and apoptosis. Cancer is now diagnosed by two different but complementary modalities, liquid biopsy, and tumor tissue biopsy. Tumor tissue biopsy is still the “gold standard” for cancer diagnosis by pathologists. ISH is ideally suited for detecting miRNA in the “gold standard” of fresh, frozen or formalin-fixed paraffin-embedded (FFPE) biopsy samples due to its sensitivity, specificity and the spatial information on tumor heterogeneity it provides, available at relatively low cost. BioGenex ISH system has the ability to generate a robust chromogenic signal while preserving the spatial context of the tissue sample. This provides a powerful tool for precise tumor characterization and a breakthrough for clinical research and analysis of cancer of unknown primary (CUP), poorly differentiated or undifferentiated tumors, and cancer staging. 

Conclusion: Only the ISH techniques give the full spatial picture of the tumor which is crucial to aid the pathologist in diagnosis and for initiating tumor treatment decisions.  

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Benchtop fully automated All-in-One ten slide system for Fluorescence in situ hybridization (FISH), ISH, miRNA and IHC

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Fluorescent in situ Hybridization (FISH) - Made Easy

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EZ-Retriever® + EZ-AR™ Elegance solution - Features and Benefits

EZ-Retriever® System:


Antigen Retrieval
- invented by BioGenex - is an effective method of unmasking antigenic epitopes in formalin-fixed paraffin-embedded tissue sections (FFPE) by heating in microwave using specially formulated Antigen Retrieval solutions. The Antigen Retrieval technique breaks the formalin induced cross-linking of proteins, allowing access of antigenic epitopes. Covered by patents issued to BioGenex, this method has been practiced in laboratories throughout the world.

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EZ-AR2 Elegance – All-in-One and All-at-Once Antigen Retrieval solution

Antigen Retrieval:

Antigen retrieval is an effective method of unmasking antigenic epitopes on the surface of formalin-fixed paraffin-embedded (FFPE) tissue sections. The antigen retrieval technique breaks the methylene bridges between epitopes and unrelated proteins to expose antigenic sites for antibody binding (1). BioGenex Laboratories Inc. invented the reverse epitope masking method in 1991 (2), with this assay now being routinely practiced in laboratories throughout the world. For in-depth article on Antigen Retrieval click here.

EZ-AR2 Elegance Antigen Retrieval Solution:

BioGenex EZ-AR2 Elegance is the first universal one-step solution for preparation of formalin-fixed paraffin-embedded (FFPE) tissue sections for IHC staining. The high throughput system is designed to perform de-wax, rehydration, and antigen retrieval all in one simple step. EZ-AR2 Elegance buffer together with EZ-Retriever® processes about 100 slides in less than 30 minutes. Efficient antigen retrieval allows higher dilution of antibodies and shortens incubation times for many antibodies – cost-effective and saves time!

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FISH: Applications in Cancer Diagnostics

In our previous article, we described about the molecular genetic technique of FISH and enumerated its many advantages in cancer diagnostics. Please refer to FISH in molecular cancer diagnostics: the whats and whys?
https://www.biogenex.com/us/blog/fish-in-molecular-cancer-diagnostics-the-whats-and-whys

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FISH in molecular cancer diagnostics: the whats and whys?

Fluorescence in situ hybridization (FISH) – a relatively new cytogenetic technique - is a DNA hybridization-based technique that generally uses directly-labeled fluorescent DNA probes to target specific chromosomal locations within the nucleus, resulting in colored signals that can be detected using a fluorescent microscope. Alternatively, FISH probes can be indirectly labeled with reporter molecules that are subsequently detected by fluorescent antibodies or other affinity molecules.

FISH is applied to detect genetic aberrations including

  • Characteristic gene fusions or translocations
  • Characteristic gene rearrangements
  • Partial or complete loss of chromosome
  • Presence of an abnormal number of chromosomes in a cell

Hence FISH can be applied to diseases covering genetic etiologies as well as cancer –both hematological and solid tumors. Additionally FISH can be applied to basic research applications like gene mapping or in discovery based assays like elucidation of novel oncogenes. Furthermore it can be used to aid in novel biomarker discovery. FISH has now been expanded to screen/analyze whole genome in one-go (in single experiment) using multicolor whole chromosome probe techniques like multiplex FISH or spectral karyotyping.

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Xmatrx® Elite – All-in-One & All-at-Once complete workflow solution! - For Your Tissue Diagnostics Needs!!

High-throughput and automation are the keywords in any cancer diagnostic lab – our Xmatrx® series are just that and more! Xmatrx Elite is versatile for any slide-based staining – IHC, ISH, FISH, multiplex IHC, in situ PCR, micro-RNA, and special staining (SS). Crisp and reliable staining result! Everytime!


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i6000 – the workhorse of a pathology lab! - (Interface for both clinical and research diagnostics)

When it comes to cancer diagnostics, tissue-based staining is still the gold standard of diagnostic workflow. And when there are thousands of cancer patients – worldwide - waiting eagerly for “their life and death question”, there is no luxury of time! Everyday there could be hundreds of samples waiting to be tested – and so some form of automation and high-throughput becomes mandatory.

Enter the semi-automatic workhorse of cancer diagnostics – the BioGenex i6000TM system. The BioGenex i6000 Elite Dx is a semi-automated, high-throughput, multiplex IHC system that is flexible, efficient and value for the bucks!

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