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Test Menu PCR

PhenoPath offers an extensive array of diagnostic assays, including over 250 IHC antibodies, over 100 flow antibodies and a broad menu of FISH, PCR, Cytogenetics and IF assays. Antibody targets include cell and tissue-specific markers, tumor prognostic and predictive markers, surrogate markers of chromosomal translocations, and other esoteric antigens.

PhenoPath employs advanced analytic instrumentation and internally developed information management technology to ensure high quality of results and rapid test turn-round. Morphometric image analysis, based on the MetaSystems platorm, is utilized in our FISH lab. GalliosTM 10-color flow cytometers support our flow laboratory. Bio-Rad, Coulter and Roche PCR platforms provide automated real-time, capillary electrophoresis and reverse transcriptase analytics. The Aperio digital slide imaging system provides digital whole slide images from our IHC laboratory to our CRO and other clients.

Our scientists employ rigorous test validation and quality control methods. Assays are validated on a variety of specimen types and are optimized to accommodate small sample size.

  • IHC and IF

    Immunohistochemistry (IHC) and immunofluorescence (IF) are immunochemical techniques employing specific antibodies along with highly sensitive detection systems that produce a colored reaction product visible under the light microscope (IHC) or fluorescence microscope (IF). IHC can be applied to either frozen or fixed tissues, whereas IF is largely restricted to frozen tissue. The power of these techniques is the simultaneous retention of tissue morphology, permitting the integration of morphologic and immunophenotypic information. PhenoPath Laboratories maintains a large and ever-expanding library of 250+ antibodies, each of which has been validated and optimized for use in diagnostic tissue-based studies.

    Advantages
    Innovative use of novel markers
    Continuously expanding menu of 250+ antibodies validated and optimized for use in clinical diagnostics
    Targets include cell- and tissue-specific markers, tumor prognostic and predictive markers, surrogate markers of chromosomal translocations, and other esoteric antigens
    Pathologists and technologists are available to answer your clinical and technical questions
    World-renowned breast cancer expertise on the forefront of standardization and clinical research
    300+ publications in the field of IHC
    PhenoPath physicians developed diagnostic antibodies for melanoma, carcinomas and infectious diseases used in testing today
    Findings correlated with histologic, immunophenotypic, molecular and clinical findings
    Rigorous test validation and quality control
    Clinically validated tests, where warranted
    Test results reported within 1 to 2 business days of receipt
    Specialized Consultative Services
    Large cell undifferentiated malignant neoplasms
    Small, blue, round cell tumors of childhood
    Carcinomas of unknown primary site
    Mesothelioma vs. adenocarcinoma
    Identification and subtyping of germ cell tumors
    Spindle cell tumor/sarcoma subtyping
    Prognostic and predictive markers in breast, lung and colon cancer
    Mature and immature lymphoid neoplasms
    Plasmacytic neoplasms
    Mature and immature myeloid neoplasms
    Amyloid subtyping
    Hydatidiform mole evaluation (IHC and FISH)
    Skin and mucosal immunofluorescence studies
    Identification of organisms, e.g., viruses, bacteria
    Validation services for ER, PR (IHC), HER2 (FISH)

     

  • Flow Cytometry

    Flow cytometry is a technique for evaluating antigen expression in suspensions of dispersed living cells. Various combinations of antibodies conjugated to fluorescent molecules, or fluorochromes, are incubated with the cell suspension according to standard protocols. The mixture of cells and bound antibodies is then introduced into the flow cytometer, and the cell-associated fluorescence in response to illumination with one or more lasers is recorded. Computer-assisted analysis of the patterns of fluorescence helps indicate whether the cells of interest are benign or malignant and, in cases of malignancy, can provide a specific diagnosis.

    While flow cytometry has historically been used to characterize hematolymphoid cell populations, this technique can be applied to any monodispersed cell suspension. At PhenoPath Laboratories, we perform up to 10-color analysis on a state-of-the-art flow cytometer, utilizing a collection of over 100 fluorescently labeled antibodies to specific antigens. Our flow cytometry lab is staffed by ASCP-certified medical technologists, representing decades of experience in multicolor flow cytometry.

    Advantages
    Extensive experience using state-of-the-art, 10-color flow, with expertise recognized by the International Clinical Cytometry Society
    Fluorescently labeled antibodies to 80+ unique antigens, plus a complete set of isotype control antibodies validated
    10-color flow yields 61% more information than 8-color, 300% more than 6-color, and 750% more than 4-color, based on the number of unique dots
    Greatly enhances the diagnostic yield of very small or paucicellular specimens (e.g., FNAs, CSFs)
    Enhanced efficiency enables rapid TAT, typically within 24 hours of specimen receipt
    When possible, > 100,000 viable cells evaluated for each flow tube, > 300,000 in post-therapy settings, enhancing our ability to identify small abnormal populations
    Pathologists and technologists available to address your clinical and technical questions

    Newly diagnosed abnormal cell populations, including all urgent cases, discussed with referring physician as needed, to ensure morphologic and clinical correlation

    Staffed by ASCP-certified medical technologists, representing decades of experience
    Rigorous test validation and quality control
    Consultative Studies
    Mature lymphoproliferative disorders
    Lymphoblastic leukemias/lymphomas
    Plasma cell dyscrasias
    Myelodysplastic syndromes
    Chronic myeloproliferative neoplasms
    Acute myeloid leukemia
    PNH
    Lymphoma staging
    Monitoring of residual disease following therapy
    Evaluating potential therapeutic targets (CD20, CD22, CD25, CD30, CD33, CD52)
    Small cell carcinoma
    Full hematopathology consultative services available

     

  • PCR

    Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA, to produce sufficient genetic material for many different types of subsequent analyses. PCR can readily identify unique DNA or RNA sequences (the latter when combined with reverse transcriptase methodologies) that are present at even a few copies per cell. It is therefore far more sensitive than methods such as CISH, FISH, and Southern and northern blotting.

    An ever-expanding number of neoplasms are being associated with specific genetic abnormalities detectable by PCR. These abnormalities may be of diagnostic, prognostic or therapeutic importance, making PCR a critical technique for the diagnostic pathology laboratory.

    PhenoPath Laboratories has validated both standard PCR coupled with capillary electrophoresis, as well as real time/quantitative PCR technologies, in a continually expanding collection of PCR assays. Our assays can be applied to fresh, frozen or paraffin-embedded tissue. Importantly, PCR findings are correlated with the histologic, immunophenotypic and clinical findings in all cases to reach a final diagnosis.

    Advantages
    Assays can be applied to fresh, frozen or FFPE tissue
    Mutation analysis via real-time PCR (1 step vs. 3 steps for sequencing), resulting in a streamlined testing process and decreased chance for errors
    Real-time PCR much more sensitive than sequencing (≤1% sensitivity vs. ~30% sensitivity)
    Rapid, sensitive and specific way to detect clonality in atypical lymphoid proliferations
    Aids in diagnosis of small biopsies involving extranodal sites (e.g., GI tract or skin)
    Correlated with histologic, immunophenotypic, molecular and clinical findings
    Reviewed and signed out by experienced, board-certified pathologists

    Pathologists and technologists available to address your clinical and technical questions

    Rigorous test validation and quality control
    Gene rearrangement assays based on landmark European Biomed-2 consortium recommendations
    Standard, capillary electrophoresis and real-time/quantitative PCR technologies validated
    Test results reported within 2 to 4 business days of receipt
    Specialized Consultative Studies
    IGH chain gene PCR to look for clonal B cell populations in the workup of B cell malignancies
    T cell receptor-gamma (TCR-gamma) gene PCR to look for clonal T cell populations in the workup of T cell malignancies
    Prognostic and predictive markers in colon, lung and melanoma
    Gene mutational analysis to aid in therapeutic decision-making
    Hematolymphoid diagnosis and subtyping
  • FISH

    Fluorescence in situ hybridization (FISH) has become an essential tool in the diagnosis and management of a variety of hematologic malignancies and non-hematopoietic tumors, as well as an aid in the identification of particular genetic disorders. FISH can be applied to fresh or fixed tissue specimens and employs DNA probes comprised of specific nucleic acid sequences attached to fluorescent molecules. These probes can be used to look for increased copies of specific genes (such as HER2 amplification in breast cancer) or entire chromosomes (such as trisomy 17 in partial hydatidiform moles), as well as decreased copies of specific genes (such as 1p and 19q in oligodendroglioma) or chromosomes (such as monosomy 7 in a subset of myelodysplastic syndromes). FISH can also identify specific chromosomal translocations, including the t(14;18) characteristic of follicular lymphoma, and the t(9;22) characteristic of chronic myeloid leukemia.

    PhenoPath Laboratories has a growing collection of validated FISH probes suitable for ancillary testing of fresh and formalin-fixed pathology specimens. FISH preparations are typically examined with a MetaSystems image analysis system that is integrated with a fluorescence microscope, permitting highly accurate and precise quantitative analysis of these genetic alterations.

    Advantages
    Morphometric image analysis using a MetaSystems image analysis system and Metafer4 application software
    Every test reviewed and signed out by an experienced, board-certified pathologist
    Pathologists and technologists available to address your clinical and technical questions
    Findings are correlated with histologic, immunophenotypic, molecular and clinical findings
    Rigorous test validation and quality control
    Assays can be applied to fresh or paraffin-embedded tissue
    Rapid turnaround time: Test results reported within 3 to 4 business days of receipt
    Tests run 5 days/week
    Specialized Consultative Services
    Prognostic and predictive markers in breast (HER2) and lung (EGFR and ALK)
    Breast HER2 testing with detailed chromosome 17 evaluation for increased accuracy of HER2 gene status
    Hydatidiform mole evaluation (FISH and IHC)
    Hematolymphoid diagnosis, prognosis and subtyping
    • B cell neoplasms
    • Myeloid neoplasms
    Small, blue, round cell tumors of childhood
    Spindle cell sarcoma/tumor subtyping
    Floater analysis by X/Y genotyping
    Validation services for HER2 FISH

     

  • Cytogenetics

    Cytogenetics is the microscopic analysis of metaphase chromosomes. The cells of interest are cultured using standard cell culture techniques to increase their number, then treated with a mitotic inhibitor to stop cell division at mitosis. Next they are centrifuged and placed in a hypotonic solution, which allows the cells to swell. The cells are then fixed, dropped on specimen slides, and the chromosomes banded, matched into pairs, and analyzed under a microscope. PhenoPath uses MetaSystems image analysis to aid in the rapid identification of metaphase cells.

    Cytogenetics surveys the entire genome for abnormalities, and can readily show deletions, duplications, and translocations. It is often performed as a complement to FISH analyses in the diagnosis of neoplastic specimens.

    Cytogenetics Studies

    • Hematologic Analysis - Bone Marrow, Blood, Lymph Node (neoplastic specimens)
    • Solid Tumor - fresh tissue

     

                                              Features/Benefits
    Provides comprehensive chromosomal studies in combination with flow, IHC, FISH and molecular studies, providing a single source solution
    Cross-disciplinary cooperation between pathologists to deliver accurate diagnoses
    Board-certified clinical cytogeneticists available for consultation
    Over 20 years of experience in diagnostic cytogenetics
    Each case analyzed by at least 2 technologists, and reviewed by a board-certified cytogeneticist
    Pathologists and technologists available to address your clinical and technical questions
    Use of image analysis (Metasystems) increases ability to identify metaphases when mitotic index is low
    Culture method optimized for increased sensitivity in detecting abnormalities
    Rapid turnaround time: neoplastic specimens: 5-7 days; solid tumor: 10-14 days

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