FISH assay for detection of HER-2 gene amplification and over-expression
The proto-oncogeneHER-2 gene (also called ERBB2 or HER2/neu) is a member of the human epidermal growth factor receptor(HER/EGFR/ERBB) family, which is locatedat the long arm of human chromosome 17 (17q12-q21).HER-2 geneencodes a 185-kDa HER-2 protein, which is a membrane receptor tyrosinekinase with homology to EGFR(HER-1).
Amplification or over-expression of HER-2 gene has been shown to play an important role in the development and progression of several aggressive types of human cancers, which occurs in 20-30% of breast cancer, approx. 20% of gastric cancer, also prostate, ovarian cancer and uterine serous endometrial carcinoma.
Demonstration of HER-2 gene amplification or over-expression is essential for initiating therapy with HER-2 targeted therapy, such as Herceptin™, a monoclonal antibody to HER-2 protein. Clinical studies have shown that patients whose tumors have high HER-2 gene amplification or over-expression benefit most from Herceptin™.
The kit is a qualitative test to accurately identify HER-2 gene amplification and over-expression via fluorescence in situhybridization (FISH) technology. The used specimen is formalin-fixed paraffin-embedded (FFPE) tissue of breast and gastric cancers.
The kit usesfluorescence in situhybridization (FISH) technology, which employs a dual-colorHER-2/CSP17 FISH probe with two fluorescent-labeled DNA probes to detect HER-2 gene amplification and over-expression.The tissue specimens were handled with a series of pre-treatment procedures, and the targeted DNA is denatured to single-stranded form and hybridized with the FISH probes. Following a series of wash andcounterstain steps, the hybridization of the probes is viewed using a fluorescence microscope equipped with appropriate excitation and emission filters, then the HER-2/CSP17 ratio is calculated.
The AmoyDx EGFR Mutations Detection Kit is a sensitive and selective assay for the detection of the most informative somatic mutations in the epidermal growth factor receptor (EGFR) gene. It is used clinically in China to select NSCLC patients who are most likely to respond to Iressa (Gefitinib) or Tarceva (Erlotinib). AmoyDx works closely with AstraZeneca in China to promote the use of molecular diagnostic tests for patient stratification.
The EGFR assay tests for a total of 29 mutations in eight PCR tubes, including both activating and resistance mutations. The presence of activating mutations indicates predicted response to mutant EGFR-targeting drugs. If the tumor tissue harbors resistance mutations, it may respond poorly to therapies designed to inhibit EGFR signalling.
The American Society for Clinical Oncology (ASCO) recommends that patients with NSCLC who are being considered for first-line therapy with an EGFR TKI (patients who have not previously received chemotherapy or an EGFR TKI) should have their tumor tested for EGFR mutations to determine whether an EGFR TKI or chemotherapy is the appropriate first-line therapy. (Keedy, V.L. et al. (2011) J. Clin. Oncol. 29(15), 2121).
The European Committee for Medicinal Products for Human Use (CHMP) recommended that EGFR mutation status be tested in patients with metastatic non-small cell lung cancer to predict sensitivity to Iressa. (EPAR summary for the public. EMEA/280173/2009. EMEA/H/C/1016).
Intended Use: CFDA approved for clinical use in China and CE marked for IVD use in Europe.
The EGFR kit employs proprietary real-time PCR technology to detect 29 mutations in the EGFR gene. Target DNA is amplified with mutation-specific PCR primers, and the mutant amplicons are detected with a novel fluorescent probe. For high quality DNA, about 2 ng of purified DNA is added to each PCR tube. For DNA from FFPE tissue older than three years, 10 to 15 ng of DNA is recommended. The test can detect mutations at a sensitivity of 1% ( i.e., one mutant copy to 99 wild-type copies).
ROS1 is a receptor tyrosine kinase of insulin receptor family. Chromosomal rearrangements involving the ROS1 receptor tyrosine kinase gene occur in a subset of non–small-cell lung cancer (NSCLC). The ROS1 fusion partners include SLC34A2, CD74, SDC4, EZR etc. These fusions lead to constitutive kinase activity and activation of downstream pathways, such as JAK/STAT, PI3K/AKT, RAS/MAPK etc., leading to carcinogenesis. It has been reported that the presence of the ROS1 rearrangement is correlated with the efficacy of TKI therapy.Based on analysis of tumor messenger RNA, ROS1 gene fusions can be detected by real-time PCR method.
Amoy Diagnostics has reached an agreement with OxOnc Development to use AmoyDx ROS1 Gene Fusions Detection Kit for a pivotal clinical trial of Pfizer's Xalkori (crizotinib) in Japan, Korea and China. Additional information is available at related news.
CE marked for IVD use in Europe.
This kit is a nucleic acid amplifcation test for the qualitative detection of ROS1 gene fusions in NSCLC FFPE samples. The kit is based on three major processes: (1) specimen preparation to isolate total RNA from NSCLC FFPE samples; (2) reverse transcription of the target RNA to generate complementary DNA (cDNA); (3) simultaneous PCR amplification of target cDNA and detection of ROS1 gene fusions with specific primers and fluorescent probes.
The AmoyDx EML4-ALK Fusion Gene Detection Kit is a sensitive and selective assay for the detection of the twenty-one most common fusion transcripts of the chimeric EML4-ALK gene.
The EML4-ALK fusion oncogene represents one of the newest molecular targets in non-small-cell lung cancer (NSCLC). First described in 2007, the fusion results from a small inversion within chromosome 2p. This inversion leads to expression of a chimeric tyrosine kinase having the N-terminal half of echinoderm microtubule-associated protein-like 4 (EML4) fused to the intracellular kinase domain of anaplastic lymphoma kinase (ALK). The EML4-ALK protein possesses potent oncogenic activity both in vitro and in vivo. This activity can be blocked by small-molecule inhibitors, supporting a role for EML4-ALK as a key driver of lung tumorigenesis.
Intended Use: CE marked for IVD use in Europe and CFDA approved for clinical use in China.
Technological Principles: The EML4-ALK kit employs proprietary real-time PCR technology to detect the twenty-one most common EML4-ALK fusion transcripts. RNA that has been purified from the FFPE tissue is first reverse transcribed to cDNA with RT enzyme supplied with the kit. The EML4-ALK fusion cDNAs are then specifically amplified with cross-junction PCR primers and quantitatively detected with fluorescent probes.
Key Features and Benefits:
• Simple Procedure — only one step required for sample application, results obtained within 90 minutes. • ADx-ARMs technology — employs a patented two-step PCR amplification procedure and novel fluorescent probe design. • Easy Standardization — any general PCR lab can perform the test, and technicians obtain reproducible results without specialized training. • Reliable product quality — Produced in a dedicated GMP plant, with quality management systems that have passed Chinese SFDA and international ISO13485 standards. • Sample type: FFPE tissue.
Detection of 16 mutations in NRAS codons 12, 13, 59, 61, 117 and 146
NRAS is a member of the RAS family of small GTPases and plays a central role in the MAPK signaling pathways. NRAS has been implicated in the pathogenesis of several cancers. NRAS mutations are particularly common in melanoma, hepatocellular carcinoma, myeloid leukemias, and thyroid carcinoma. In total, activating mutations in the NRAS genes occur in 13~25% of cutaneous melanomas, 1~6% of colorectal cancer and 1% of lung cancer, mainly in exons 2, 3 or 4. The mutation status of NRAS gene is relevant to drug resistance of non-small cell lung cancer treated with tyrosine kinase inhibitors. As a result, NRAS mutations detection supplies evidence for targeted clinical therapy of tumor patients.
AmoyDx® NRAS Mutation Detection Kit is CE marked for IVD use in Europe.
The kit uses novel, patented primers and probes to detect mutations in a real-time PCR assay. The mutant DNA is amplified accurately by the specific primers, and detected by the novel probes. Highly Specific primers and probes, and a highly validated procedure based on Taq DNA polymerase contribute to outstanding assay sensitivity and selectivity.
Amoy Diagnostics provides state-of-the-art molecular diagnostic products and services to healthcare providers, academic institutions, and pharmaceutical companies. The company has a rich portfolio of tests that focuses on detecting mutations and measuring gene expression in FFPE cancer tissue. The assays are based on a proprietary real-time PCR technology called “ADx-ARMS”. They have outstanding sensitivity (1%) and accuracy (99%). Three key tests – for mutations in the EGFR, KRAS and BRAF genes - have received both CE declarations for marketing in Europ...