KIT Asp816Val Mutation Analysis, Qualitative PCR
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
Diagnosing systemic mastocytosis
Testing Algorithm Delineates situation(s) when tests are added to the initial order. This includes reflex and additional tests.
Special Instructions and Forms Describes specimen collection and preparation information, test algorithms, and other information pertinent to test. Also includes pertinent information and consent forms to be used when requesting a particular test
Allele-Specific Oligonucleotide Polymerase Chain Reaction (PCR)
(PCR is utilized pursuant to a license agreement with Roche Molecular Systems, Inc.)
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
KIT Asp816Val Mutation Analysis, V
Systemic Mastocytosis Mutation
Systemic Mastocytosis Mutation
Specimen Type Describes the specimen type needed for testing
Specimen Required Defines the optimal specimen. This field describes the type of specimen required to perform the test and the preferred volume to complete testing. The volume allows automated processing, fastest throughput and, when indicated, repeat or reflex testing.
The following information is required:
1. Pertinent clinical history
2. Clinical or morphologic suspicion
3. Date of collection
4. Specimen source
1. Hematopathology Patient Information Sheet (Supply T676) in Special Instructions
2. If not ordering electronically, submit a Hematopathology/Molecular Oncology Request Form (Supply T241) with the specimen.
Specimen must arrive within 168 hours of collection.
Submit only 1 of the following specimens:
Specimen Type: Paraffin-embedded bone marrow aspirate clot
Container/Tube: Paraffin block
Specimen Stability Information: Ambient/Refrigerated
Specimen Type: Extracted DNA from blood or bone marrow
Container/Tube: 1.5- to 20-mL tube with indication of volume and concentration of DNA
Specimen Volume: Entire specimen
Collection Instructions: Label specimen as extracted DNA from blood or bone marrow
Specimen Stability Information: Refrigerated/Ambient
Specimen Minimum Volume Defines the amount of specimen required to perform an assay once, including instrument and container dead space. Submitting the minimum specimen volume makes it impossible to repeat the test or perform confirmatory or perform reflex testing. In some situations, a minimum specimen volume may result in a QNS (quantity not sufficient) result, requiring a second specimen to be collected.
Extracted DNA from Blood or Bone Marrow: 50 mcL at 20 ng extracted DNA/mcL
Bone marrow biopsies, slides, or paraffin shavings
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Systemic mastocytosis is a hematopoietic neoplasm that can be included in the general category of chronic myeloproliferative disorders (CMPDs). These neoplasms are characterized by excessive proliferation of 1 or more myeloid lineages, with cells filling the bone marrow and populating other hematopoietic sites. In systemic mastocytosis, mast cell proliferation is the defining feature, although other myeloid lineages and B-cells are frequently part of the neoplastic clone.
Function-altering point mutations in KIT, a gene coding for a membrane receptor tyrosine kinase, have been found in myeloid lineage cells in the majority of systemic mastocytosis cases. The most common KIT mutation is an adenine-to thymine base substitution (A->T) at nucleotide position 2468, which results in an aspartic acid-to-valine change in the protein (Asp816Val). Much less frequently, other mutations at this same location are found and occasional cases with mutations at other locations have also been reported. Mutations at the 816 codon are believed to alter the protein such that it is in a constitutively activated state. The main downstream effect of KIT activation is cell proliferation.
Detection of a mutation at the 816 codon is included as 1 of the minor diagnostic criteria for systemic mastocytosis in the World Health Organization (WHO) classification system for hematopoietic neoplasms and is also of therapeutic relevance, as it confers resistance to imatinib, a drug commonly used to treat CMPDs. It is now clear that individual mast cell neoplasms are variable with respect to the number of cell lineages containing the mutation; some having positivity only in mast cells and others having positivity in additional myeloid and even lymphoid lineages. The mutation has not been reported in normal tissues.
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
An interpretive report will be provided indicating the mutation status as positive or negative.
The test will be interpreted as positive or negative for KIT Asp816Val.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
Some systemic mastocytosis cases may have the mutation only in mast cells. Since these cells rarely circulate in blood and are difficult to obtain in significant numbers from bone marrow aspirate specimens, false-negative results may occur if neoplastic cells are present below the sensitivity of the assay (fewer than 0.01% mutated alleles).
The test is qualitative only. Reliable quantitative results cannot be issued.
The analytic sensitivity of this test is 0.01% and was determined by the dilution of a cell line containing homozygous KIT mutation. This means that 0.01% or greater of the KIT alleles present in the specimen must contain the mutation to be detected by the assay. The analytic specificity was 100% in assay validation.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Garcia-Montero A, Jara-Acevedo M, Teodosio C, et al: KIT mutation in mast cells and other bone marrow hematopoietic cell lineages in systemic mast cell disorders: a prospective study of the Spanish Network on Mastocytosis (REMA) in a series of 113 patients. Blood 2006;108-2366-232
2. Valent P, Akin C, Sperr WR, et al: Diagnosis and treatment of systemic mastocytosis: state of the art. Br J Haematol 2003;122:695-717
3. Jaffe ES, Harris NL, Stein H, et al: World Health Organization Classification of Tumours. Pathology and Genetics. Tumours of the Haematopoietic and Lymphoid Tissues. 2001, pp 291-302
Method Description Describes how the test is performed and provides a method-specific reference
The KIT mutation assay developed for clinical use in the Mayo Molecular Hematopathology Laboratory detects the KIT mutation responsible for Asp816Val. The technique used is allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) with fragment analysis on an ABI3100 genetic analyzer. Briefly, DNA is extracted from whole bone marrow or blood and PCR is used to amplify across the mutation site in 2 separate tubes; 1 contains a reverse primer complementary to the unmutated sequence and the other contains a reverse primer complementary to the mutated sequence. Each of these reverse primers is labeled with a fluorescent tag and both tubes contain an identical, nonlabeled forward primer. Both primer sets amplify a 200 bp fragment that differs only at the mutation site. The unmutated fragment should be amplified in all samples. Samples negative for KIT Asp816Val will not have an amplified fragment in the mutated reaction tube. Positive samples will have amplified fragments in both the unmutated and mutated tubes. The test gives a qualitative (positive or negative) result only, as the end point PCR used is not reliable for quantification.(Unpublished Mayo method)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test
Test Classification Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer's instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR), Investigation Use Only (IUO) product, or a Research Use Only (RUO) product.
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.
CPT Code Information Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Medical Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.
81402-KIT (v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog) (eg, mastocytosis), common variants (eg, D816V, D816Y, D816F)
LOINC® Code Information Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the result codes returned for this test or profile.
|Result ID||Reporting Name||LOINC Code|
|MP010||Specimen Type||In Process|