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Epic Code LAB848 Galactosemia Reflex, Blood

Important Note

Order as Reference Miscellaneous #LAB848; indicate complete test name when ordering.

Additional Codes

Mayo Code: GCT

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Useful For

Preferred test for diagnosis, carrier detection, and determination of genotype of galactose-1-phosphate uridyltransferase deficiency, the most common cause of galactosemia

 

Differentiating Duarte variant galactosemia from classic galactosemia

 

Confirming results of newborn screening programs

Specimen Type

Whole Blood EDTA


Ordering Guidance


This test is appropriate for the diagnosis of and routine carrier screening for galactose-1-phosphate uridyltransferase (GALT) deficiency.

 

This assay is not appropriate for monitoring dietary compliance. For dietary monitoring, order GAL1P / Galactose-1-Phosphate, Erythrocytes.

Necessary Information


Patient's age is required.



Specimen Required


Multiple whole blood tests for galactosemia can be performed on one specimen. Prioritize order of testing when submitting specimens. See Galactosemia-Related Test List for a list of tests that can be ordered together.

 

Container/Tube:

Preferred: Lavender top (EDTA)

Acceptable: Green top (sodium heparin) or yellow top (ACD)

Specimen Volume: 5 mL


Specimen Minimum Volume

2 mL

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:

-Informed Consent for Genetic Testing (T576)

-Informed Consent for Genetic Testing-Spanish (T826)

2. If not ordering electronically, complete, print, and send an Inborn Errors of Metabolism Test Request (T798) with the specimen.

Specimen Stability Information

Specimen Type Temperature Time Special Container
Whole Blood EDTA Refrigerated (preferred) 28 days
  Ambient  14 days

Reject Due To

Gross hemolysis Reject

Cautions

This assay will not reliably detect deletions/insertions (delins) of 40 or more base pairs (bp), including Alu insertions, long interspersed elements (LINES), and short interspersed elements (SINES). The bioinformatics software pipeline is verified to detect 95% of deletions up to 75 bp and insertions up to 47 bp.

 

Additionally, low level mosaic variants may not be detected.

 

This test is not designed to differentiate between somatic and germline variants. If there is a possibility that any detected variant is somatic, additional testing may be necessary to clarify the significance of results.

 

Test results should be interpreted in the context of clinical findings, family history, and other laboratory data. Errors in the interpretation of results may occur if information given is inaccurate or incomplete.

 

Many disorders may present with symptoms similar to those associated with galactosemia. Therefore, biochemical testing is performed to establish the diagnosis of galactosemia prior to DNA analysis.

Clinical Information

Galactosemia is an autosomal recessive disorder that results from a deficiency of any 1 of the 4 enzymes catalyzing the conversion of galactose to glucose: galactose-1-phosphate uridyltransferase (GALT), galactokinase (GALK), uridine diphosphate galactose-4-epimerase (GALE), and galactose mutarotase (GALM). GALT deficiency is the most common cause of galactosemia and is often referred to as classic galactosemia. The complete or near-complete deficiency of GALT enzyme is life threatening if left untreated. Complications in the neonatal period include failure to thrive, liver failure, sepsis, and death.

 

Galactosemia is treated by a galactose-restricted diet, which allows for rapid recovery from the acute symptoms and a generally good prognosis. Despite adequate treatment from an early age, individuals with galactosemia remain at increased risk for developmental delays, speech problems, and abnormalities of motor function. Female patients with galactosemia are at increased risk for premature ovarian failure. Based upon reports by newborn screening programs, the frequency of classic galactosemia in the United States is approximately 1 in 30,000, although literature reports range from 1 in 10,000 to 1 in 60,000 live births.

 

Galactose-1-phosphate (Gal-1-P) accumulates in the erythrocytes of patients with galactosemia. The quantitative measurement of Gal-1-P is useful for monitoring compliance with dietary therapy. Gal-1-P is thought to be the causative factor for development of liver disease in these patients and, because of this, patients should maintain low levels and be monitored on a regular basis.

 

Duarte-variant galactosemia (compound heterozygosity for the Duarte variant, N314D, and a classic variant) is generally associated with higher levels of enzyme activity (5%-20%) than classic galactosemia (<5%); however, this may be indistinguishable by newborn screening assays. Previously, it was unknown whether children with Duarte-variant galactosemia were at an increased risk for adverse developmental outcomes due to milk exposure so were often treated with a low galactose diet during infancy. More recently however, the outcomes data suggest a lack of evidence for developmental complications due to milk exposure, therefore treatment recommendations remain controversial. The Los Angeles variant, which consists of N314D and a second genetic variant, L218L, is associated with higher levels of GALT enzyme activity than the Duarte-variant allele.

 

Newborn screening for galactosemia is performed in all 50 US states, though the method by which potentially affected individuals are detected varies from state to state and may include the measurement of total galactose (galactose and Gal-1-P) and/or determining the activity of the GALT enzyme. The diagnosis of galactosemia is established by follow-up quantitative measurement of GALT enzyme activity. If enzyme level is less than 24.5 nmol/h/mg of hemoglobin, sequencing of the GALT gene is performed.

 

For more information see Galactosemia Testing Algorithm.

Method Name

Enzyme Reaction followed by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Report Available

4 to 5 days

Reporting Name

Galactosemia Reflex, B

Reference Values

≥24.5 nmol/h/mg of hemoglobin

Interpretation

The laboratory provides an interpretation of the results, including galactose-1-phosphate uridyltransferase enzyme activity and genotype, if necessary. This interpretation provides an overview of the results and their significance, a correlation to available clinical information, elements of differential diagnosis, and recommendations for additional testing.

 

In any specimen where enzyme activity is less than 24.5 nmol/h/mg of hemoglobin GALT full gene sequencing will be performed. For testing algorithm and more information, see Galactosemia Testing Algorithm.

 

The GALT gene maps to chromosome 9p13. Several disease-causing variants are common in patients with classic galactosemia (G/G genotype). The most frequently observed is the Q188R classic variant. This alteration accounts for 60% to 70% of classic galactosemia alleles. The S135L variant is the most frequently observed in African Americans and accounts for approximately 50% of the altered alleles in this population. The K285N variant is common in those of eastern European descent and accounts for 25% to 40% of the alleles in this population. The L195P variant is observed in 5% to 7% of classical galactosemia. The 5-kilobase deletion is common in individuals of Ashkenazi Jewish descent. The Duarte variant (N314D and -119_-116delGTCA) is observed in 5% of the general US population.

Method Description

Galactose-1-Phosphate Uridyltransferase Enzyme Analysis:

An aqueous mixture containing water, uridine diphosphate (UDP)-glucose, (13)C(2)-labeled galactose-1-phosphate, and UDP-N-acetylglucosamine (internal standard) is added to hemolysate aliquot. The mixture is then vortexed briefly and incubated.

 

After incubation the reaction is quenched, extracted, and centrifuged. The top layer is then transferred to a 96-well plate, and then injected onto a liquid chromatography-tandem mass spectrometry (LC-MS/MS). The ratio of the extracted peak area of (13)C2 labeled UDP-galactose to its internal standard UDP-N-acetylglucosamine as determined by LC-MS/MS is used to calculate the concentration of product analyte in the sample. The concentration of the product is then normalized using the calculated hemoglobin concentration to determine the patient's enzyme level in nmol/h/mg of hemoglobin.(Unpublished Mayo method)

 

GALT Full Gene Sequencing:

Next-generation sequencing (NGS) and/or Sanger sequencing is performed to test for the presence of variants in coding regions and intron/exon boundaries of the GALT gene. NGS and/or a polymerase chain reaction (PCR)-based quantitative method is performed to test for the presence of deletions and duplications in the GALT gene.

 

There may be regions of GALT that cannot be effectively amplified for sequencing or deletion and duplication analysis because of technical limitations of the assay, including regions of homology, high guanine-cytosine (GC) content, and repetitive sequences. Confirmation of select reportable variants may be performed by alternate methodologies based on internal laboratory criteria.

 

PCR-based methods and/or Sanger sequencing is used to confirm variants detected by NGS when appropriate.(Unpublished Mayo method)

Day(s) Performed

Monday, Wednesday, Friday

Clinical Reference

1. Berry GT: Classic galactosemia and clinical variant galactosemia. In: Adam MP, Ardinger HH, Pagon RA, et al, eds. GeneReviews [Internet]. University of Washington, Seattle; 2000. Updated March 11, 2021. Accessed April 12, 2022. Available at www.ncbi.nlm.nih.gov/books/NBK1518/

2. Walter JH, Fridovich-Keil JL: Galactosemia. In: Valle D, Antonarakis S, Ballabio A, Beaudet AL, Mitchell GA. eds. The Online Metabolic and Molecular Bases of Inherited Disease. McGraw-Hill; 2019 Accessed April 12, 2022. Available at https://ommbid.mhmedical.com/content.aspx?bookid=2709&sectionid=%20225081023

3. Carlock G, Fischer ST, Lynch ME, et al: Developmental outcomes in Duarte galactosemia. Pediatrics. 2019 Jan;143(1):e20182516. doi:10.1542/peds.2018-2516

Test Classification

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

82775

81406 (if appropriate)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
GCT Galactosemia Reflex, B 24082-0

 

Result ID Test Result Name Result LOINC Value
8333 Gal-1-P Uridyltransferase, RBC 24082-0
2296 Interpretation (GALT) 59462-2
58115 Reviewed By 18771-6

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
GALZ Galactosemia, Full Gene Analysis Yes No