|
|
|
| Home > Our Services > Reference Laboratory > Test Offered List > Specical Technology |
|
|
It is a technique that allows creating millions of precise DNA replications from a single sample of DNA.
The reaction requires the presence of:
Isolated genomic DNA or complementary DNA (RNA that has undergone in vitro reverse transcription to DNA)
Essential components of DNA synthesis, including deoxynucleoside triphosphates (dNTPs), specifically designed oligonucleotide primers that flank the DNA segment of interest, thermostable DNA Polymerase, an appropriate buffer that contains magnesium chloride, and specific temperature cycling parameters that permit and control the amplification process.
After cycling is completed, the amplification products can be examined in various ways. Typically, the contents of the reaction vessel are subjected to gel electrophoresis. This allows visualization of the amplified gene segments (e.g., PCR products, bands) and a determination of their specificity.
PCR can be used to detect cancers such as leukaemia and lymphomas. In fact, PCR is particularly sensitive to detection of malignant cells - far more than other techniques currently used.
Mutations can also be detected via this method of DNA analysis |
Flow Cytometry is a technique to quantify the fluorescence and light scatter of particles in suspension.
In the clinical laboratory, the particles are usually cells that fluoresce after binding to specific dyes, often fluorescently labeled antibodies.
The identification and examination of key cell signaling events required for initiation and progression of cancer might be best accomplished at the single cell level. Recent advances in flow cytometry have enabled simultaneous measurement of cell type and signaling pathways. The advantage of diagnosing a patient's cancer at the single cell level provides an approach for early detection of cancer and yield insights into how cancer cells are responding or adapting to therapy. A byproduct of the single cell technique, when appropriately extended, is that one should eventually be able to predict those pathways cancer cells might be using to circumvent current therapies and/or direct the patient towards alternative treatments.
Flow cytometry has now become the preferred method for
(1) lineage assignment,
(2) maturational characterization of malignant cells,
(3) detection of clonality,
(4) heterogeneity and aberrant features of the malignant cell populations, and
(5) quantitation of hematopoietic cells.
Triesta offers about 25 flurochrome tagged monoclonal antibodies for categorizing the acute leukemias and classify the chronic lymphomproliferative disorder in luekemic phase. |
| Immunohistochemistry is a method of analysing and identifying cell types based on the binding of antibodies to specific components of the cell. Immunohistochemistry is used to diagnose the type of cancer and to help determine the patient's prognosis. In cases such as metastases or carcinoma of unknown primary origin, where it may be difficult to determine the type of cell from which the tumor originated, immunohistochemistry can identify cells by the characteristic markers on the cell surface. IHC can also help distinguish between benign and malignant tumours. |
Karyotyping is a test to examine chromosomes in a sample of cells, which can help identify genetic problems as the cause of a disorder or disease. This test can:
Count the number of chromosomes
Look for structural changes in chromosomes
Cytogenetic studies assist physicians to look for the cause of Constitutional, Induced, Acquired abnormalities in the patients. Clinical Cytogenetic analyses are performed on stained metaphase chromosomes to produce G banding specific to each chromosome; this allows for the detection of subtle changes in Chromosome structure. Varieties of other staining techniques are available to identify specific abnormalities.
Gene mutations are not only responsible for rare hereditary forms of human cancer, but for the sporadic forms of human malignancies as well. Many of these specific genetic defects in cancer cells can be visualized as chromosomal aberrations. Conventional cytogenetic analysis of metaphase chromosomes from human malignancies is a first screening step to identify chromosomal aberrations. Since the introduction of chromosome banding techniques significant knowledge of chromosomal aberrations especially in hematological malignancies as well as sarcomas has been gained. In these malignancies, specific balanced translocations were identified and have led to the cloning of the genes involved at many breakpoints. These aberrations have proven to be of significant etiologic, diagnostic, prognostic, as well as therapeutic relevance, especially in leukemias.
Triesta offers
• Conventional Karyotyping by G banding
• Fish for BCR-ABL – t(9:22) (q34;q11) |
FISH is an important molecular cytogenetic method for identifying chromosomes and parts of chromosomes, deciphering chromosome rearrangements, and locating genes on chromosomes.
"Fluorescent" means emitting light that comes from a reaction within the emitter. "In situ" refers to the fact that this techniques is done with the chromosomes, cells or tissue in place (in situ) on a microscope slide.
In situ hybridization is a technique in which single-stranded nucleic acids (DNA or RNA) are permitted to interact so that complexes, or hybrids, are formed by molecules with sufficiently similar, complementary sequences. Through nucleic acid hybridization, the degree of sequence identity can be determined and specific sequences detected and sometimes those sequences can be located on a specific chromosome. A nucleic acid labelled with a fluorescent dye is hybridized to suitably prepared cells or histological sections. This is used particularly to look for specific transcription or localization of genes to specific chromosomes.
FISH is an essential tool in the diagnosis and management of a variety of solid tumors and hematological malignancies as well as an aid in the identification of particular genetic disorders |
|
|
|
|
