Test Catalog

Test Id : PKLRG

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Pyruvate Kinase Liver and Red Blood Cell (PKLR), Full Gene Sequencing and Large Deletion Detection, Varies

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Overview

Test Catalog

Useful For
Suggests clinical disorders or settings where the test may be helpful

Aid in the diagnosis of pyruvate kinase (PK) deficiency

 

Ascertain a causative variant in the PKLR gene of patients with low or relatively low levels of erythrocytic PK enzymatic activity

 

Ascertain carrier status of family members of individuals diagnosed with PK deficiency for genetic counseling purposes

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Name
A short description of the method used to perform the test

Polymerase Chain Reaction (PCR) followed by DNA Sequence Analysis/Large Deletion Detection by PCR followed by Fragment Analysis

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

PKLR Full Gene and Deletion

Aliases
Lists additional common names for a test, as an aid in searching

PK

Pyruvate Kinase (RBC)

Hemolytic anemia

Nonspherocytic hemolytic anemia

Specimen Type
Describes the specimen type validated for testing

Varies

Ordering Guidance

Preliminary screening tests, such as complete blood count with peripheral smear, direct Coombs test, and pyruvate kinase (PK) enzyme activity assays (preferably as a part of EEEVP / Red Blood Cell [RBC] Enzyme Evaluation) should be run before ordering this evaluation.

Necessary Information

1. PKLR Gene Sequencing Patient Information is required, see Special Instructions. Testing may proceed without the patient information however it aids in providing a more thorough interpretation. Ordering providers are strongly encouraged to complete the form and send it with the specimen.

2. Include physician name and phone number with specimen.

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Submit only 1 of the following specimens:

 

Specimen Type: Whole blood

Container/Tube: Yellow top (ACD solution B) or Purple top (EDTA)

Specimen Volume: 3 mL

Collection Instructions:

1. Invert several times to mix blood.

2. Send specimen in original tube.

Specimen Stability Information: Refrigerated 30 days

 

Specimen Type: DNA

Container/Tube: 2 mL screw top tube

Specimen Volume: 100 microliters

Collection Instructions:

1. The preferred volume is 100 microliters at a concentration of 250 ng/mcL

2. Include concentration and volume on tube

Specimen Stability Information: Frozen preferred; Ambient/Refrigerate acceptable

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Forms

1. New York Clients-Informed consent is required. Document on the request form or electronic order that a copy is on file. The following documents are available in Special Instructions:

-Informed Consent for Genetic Testing (T576)

-Informed Consent for Genetic Testing-Spanish (T826)

2. PKLR Gene Sequencing Patient Information is required, see Special Instructions

3. If not ordering electronically, complete, print, and send a Benign Hematology Test Request Form (T755) with the specimen.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

Whole blood: 0.5 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

All specimens will be evaluated at Mayo Clinic Laboratories for test suitability.

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Varies Varies (preferred)

Useful For
Suggests clinical disorders or settings where the test may be helpful

Aid in the diagnosis of pyruvate kinase (PK) deficiency

 

Ascertain a causative variant in the PKLR gene of patients with low or relatively low levels of erythrocytic PK enzymatic activity

 

Ascertain carrier status of family members of individuals diagnosed with PK deficiency for genetic counseling purposes

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

The glycolytic pathway is used by all tissues for energy production through the formation of adenosine triphosphate (ATP). It is particularly important in red blood cells, which are dependent upon this pathway for energy due to their lack of mitochondria. The PKLR gene encodes for pyruvate kinase, the rate-limiting glycolytic enzyme that catalyzes the transphosphorylation from phosphoenolpyruvate (PEP) to adenosine diphosphate (ADP) creating pyruvate and ATP. Pyruvate kinase (PK) deficiency is a relatively common cause of hereditary nonspherocytic hemolytic anemia,(1) with an estimated prevalence of 1:20,000 among people of European descent. The severity of hemolysis varies from fully compensated forms to life-threatening neonatal anemia requiring transfusions.(2) Over 200 different variants have been reported in the PKLR gene. Most are single nucleotide substitutions although rarer large deletions have also been identified. PK deficiency is inherited in an autosomal recessive manner, and genetic results should be correlated with enzyme levels performed remote from transfusion when possible. PK deficiency can be difficult to interpret based on enzyme level alone and may be only mildly decreased or normal in those with the most severe symptoms or after splenectomy due to reticulocytosis.(2) Comparison to other erythrocyte enzyme levels is usually very helpful in this regard. Heterozygous carriers of PKLR variants have intermediate enzyme levels and are not symptomatic.

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.

Interpretation
Provides information to assist in interpretation of the test results

All detected alterations will be evaluated according to current ACMG recommendations.(3) Variants will be classified based on known, predicted, or possible effect on gene pathogenicity and reported with interpretive comments detailing their potential or known clinical significance.

Cautions
Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

Blood samples may contain donor DNA if obtained from patients who received heterologous blood transfusions or allogeneic blood or marrow transplantation. Results from samples obtained under these circumstances may not accurately reflect the recipient's genotype.

 

For individuals who have received blood transfusions, the genotype usually reverts to that of the recipient within 6 weeks. For individuals who have received allogeneic blood or marrow transplantation, a pretransplant DNA specimen is recommended for testing. For patients who have been transfused within the preceding 6 weeks, the enzyme assay (PK1 / Pyruvate Kinase Enzyme Activity, Blood) will also be affected, so it is not an appropriate alternative test. 

 

Patients who have received an allogeneic blood or marrow transplant would be expected to convert to the PKLR status of the donor. However, if the patient's transplant was partially successful or if there is a relapse of an underlying hematologic malignancy, a mixture of donor and recipient genotype may be seen on genetic analysis. The enzyme assay can be run after transplantation; order PK1 / Pyruvate Kinase Enzyme Activity, Blood.

 

Rare variants exist that could lead to false-negative or false-positive results. Other variants in the primer binding regions can affect the testing, and ultimately, the genotype assessment made.

 

Test results should be interpreted in the context of clinical findings, family history, and other laboratory data. Large deletions or rearrangements that are not within the intron 2 through exon 11 region are not detected by this assay.

 

Sometimes a genetic alteration of unknown significance may be identified. In this case, testing of appropriate family members may be useful to determine pathogenicity of the alteration.

 

This test is not designed to provide specific dosing or drug selection recommendations and is to be used as an aid to clinical decision making only. Drug-label guidance should be used when dosing patients with medications regardless of the predicted phenotype.

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. van Wijk R, Huizinga E, van Wesel A, et al: Fifteen novel mutations in PKLR associated with pyruvate (PK) deficiency: structural implications of amino acid substitutions in PK. Hum Mutat. 2009;30(3):446-453

2. Zanella A, Fermo E, Bianchi P, et al: Pyruvate kinase deficiency: the genotype-phenotype association. Blood Rev. 2007;21:217-231

3. Richards S, Aziz N, Bale S, et al: Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med. 2015 May;17(5):405-424

4. OMIM: 609712 Pyruvate Kinase, Liver and Red Blood Cell; PKLR. Johns Hopkins University; 2005. Updated November 2016. Accessed March 11, 2021. Available at www.omim.org/entry/609712

5. Baronciani L, Beutler E: Molecular study of pyruvate deficient patients with hereditary nonspherocytic hemolytic anemia. J Clin Invest. 1995 April;95:1702-1709

6. Bianchi P, Zanella A: Hematologically important mutations: red cell pyruvate kinase (Third update). Blood Cells Mol Dis. 2000 Feb;26(3):47-53

7. Costa C, Albuisson J, Le TH, et al: Severe hemolytic anemia in a Vietnamese family, associated with novel mutations in the gene encoding for pyruvate kinase. Haematologica. 2005;90:25-30

8. So CC, Tang M, Li CH, et al: First reported case of prenatal diagnosis for pyruvate kinase deficiency in a Chinese family. Hematology. 2011;16(6):377-379

9. van Wijk R, van Solinge WW, Nerlov C, et al: Disruption of a novel regulatory element in the erythroid-specific promoter of the human PKLR gene causes severe pyruvate kinase deficiency. Blood. 2003 Feb;101(4):1596-1062

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Description
Describes how the test is performed and provides a method-specific reference

Genomic DNA is extracted from whole blood. The PKLR gene is amplified by polymerase chain reaction (PCR). The PCR products are then purified and sequenced in both directions using fluorescent dye-terminator chemistry. Sequencing products are separated on an automated sequencer and trace files analyzed for variations in the exons and intron/exon boundaries of all exons using variant detection software and visual inspection.(Unpublished Mayo method)

 

Large Deletion Detection:

A single PCR product is amplified and separated by gel electrophoresis for fragment size detection.(Unpublished Mayo method)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Varies

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

3 to 7 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

Whole Blood: 2 weeks; Extracted DNA: 2 months

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

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 instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) 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 US 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 Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

81405-PKLR

LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
PKLRG PKLR Full Gene and Deletion 94212-8
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
37858 Interpretation 69047-9
37860 Reviewed by 18771-6
48398 Result Details 82939-0
48396 Disclaimer 62364-5
48397 Method 85069-3
37857 Result Summary 50397-9
91971 Additional Information 48767-8

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Create a PDF

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports