Department of Molecular Genetics & Genomics

Patient information Ref. Doctor/ Hospital/ Specimen

Patient Name: Ref Hospital:
Visit No: Ref Doctor:
Barcode: Sample type:
Age/Gender: Received date:
Test name: Reported date:

Referral Reason: Genetic screening for blood clotting proteins (MTHFR, Factor V and II).

Result: Compound Heterozygous for the targeted MTHFR gene variants.

Gene Exon no Genomic Position Variant & protein change Zygosity

MTHFR chr1:11796321 c.665C>T (p. Ala222Val)
5 HETEROZYGOUS
(NM_005957.4) (GRCh38.p13) (rs1801133)
MTHFR Chr1: 11794419 c.1298A>C (p. Glu429Ala)
8 HETEROZYGOUS
(NM_005957.4) (GRCh38.p13) (rs1801131)
F5 chr1:169549811
c.1601G>A (p. Arg534Gln)
(NM_00130.4) 10 (GRCh38.p13) Normal
(rs6025)
F2 chr11:46739505
3’ UTR c.*97G>A (G20210A) Normal
(NM_000506.3) GRCh38.p13

Interpretation:
This individual is found to be COMPOUND HETEROZYGOUS for the MTHFR variants (c.665C>T) and (c.1298A> C)
in exon 5 and 8 of MTHFR gene.

Heterozygous MTHFR C677T mutation detected in exon 5 of MTHFR gene

Reference Sequence

Patient Sequence

6-3-862/A, Lal Bungalow, Ameerpet, Hyderabad – 500016, info@lifelinediagnostics.in,

Ph: +91 9542129919
Heterozygous MTHFR A1298 C mutation detected in exon 8 of MTHFR gene

Reference Sequence

Patient Sequence

Recommendations:
• Clinical correlation and Genetic counseling are recommended.
• Parental carrier testing.
• Biochemical analysis of Plasma homocysteine.

Methodology: PCR followed by bi-directional Sanger Sequencing was performed from the isolated DNA to
identify the targeted common variants in the MTHFR, factor II and factor V.

Clinical Information:
MTHFR Polymorphisms C677T & A1298C Alleles:
The C665T allele is characterised by a point mutation at position 665(677 is old nomenclature) of the MTHFR
gene that converts a cytosine (C) into a thymine (T); this mutation results in an amino acid substitution
(alanine to valine) in the enzyme. This genetic change leads to a defective MTHFR enzyme that maintains only
50% of the normal activity. The wild-type genotype (665CC) exists in 75%, while heterozygosity (677CT)
exists in 10% of the general white population from northern European descent. Homozygosity for the
valine variant (677TT) is present in about 15% of the general population and these individuals tend to
have increased blood homocysteine levels. Those heterozygous (677CT) have intermediate
homocysteine levels. The effect of the MTHFR genotype on homocysteine concentrations is most significant
among those with low folate status.
Another common polymorphism, the A1298C allele, is characterised by a point mutation at position 1298 of
the MTHFR gene that converts alanine into cytosine. This mutation results in an amino acid substitution
glutamate to alanine in the enzyme. People who are heterozygous for both alleles (665 and 1298) tend to have
increased serum homocysteine levels.
Factor V Leiden (FVL) and prothrombin gene mutation (PGM) are common inherited thrombophilia. The
c.*97G>A variant in the F2 gene is a genetic risk factor for venous thromboembolism. Heterozygous carriers
have a 2- to 4-fold increased risk for venous thromboembolism. Homozygous subjects for the c.*97G>A variant
are rare. Individuals who carry both a *97G>A variant in the F2 gene and a c.1601G>A (p. Arg534Gln) variant in
the F5 gene (commonly referred to as Factor V Leiden) have an approximately 20-fold increased risk for
venous thromboembolism. Risks are likely to be higher in more complex genotype combinations involving the
F2 c.*97G>A variant and Factor V Leiden.2 Additional risk factors include but are not limited to: deficiency of
protein C, protein S, or antithrombin III, age, male sex, personal or family history of deep vein
6-3-862/A, Lal Bungalow, Ameerpet, Hyderabad – 500016, info@lifelinediagnostics.in,
Ph: +91 9542129919
 thromboembolism, smoking, surgery, prolonged immobilization, malignant neoplasm, tamoxifen treatment,
raloxifene treatment, oral contraceptive use, hormone replacement therapy, and pregnancy. This result
cannot predict the occurrence or recurrence of a thrombotic event.

Disclaimer:
• Test results should be interpreted in the context of clinical findings, family history, and other laboratory data.
Errors in our interpretation of results may occur if information give n is inaccurate or incomplete.
• This test doesn’t cover other regions of the MTHFR, Factor V and II other than targeted variants.
• Although all precautions are taken while conducting the tests, there is a standard error rate of approximate
1% in all genetic tests and this should be taken into consideration before any clinical decision. 1
• It is presumed that the specimen used to perform the test belongs to the patient specified above, such
verification having been carried out at the collection level of sample.

References:

1. Katharina J Hoff (2009): The effect of sequencing errors on metagenomic gene prediction. BMC Genomics, 10:520.
2. Ulrich CM, Yasui Y, Storb R, Schubert MM, Wagner JL, Bigler J, Ariail KS, Keener CL, Li S, Liu H, Farin FM, Potter JD Blood.
2001 Jul 1; 98(1):231-4.
3. Levin BL, Varga E. MTHFR: addressing genetic counseling dilemmas using evidence-based literature. J Genet Couns.
2016;25(5):901-911.
4. Greenland P, Alpert JS, Beller GA, et al. 2010 ACCF/AHA guideline for assessment of cardiovascular risk in asymp tomatic
adults: a report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice
Guidelines. J Am Coll Cardiol. 2010;56(25): e50-103.
5. Coppola A, Tufano A, Cerbone AM, and Di Minno G. Inherited thrombophilia: implications for prevention and treatment
of venous thromboembolism. Semin Thromb Hemost 2009, 35:683-94. 20013535
6. Segal JB, et al. Predictive value of factor V Leiden and prothrombin G20210A in adults with venous thromboembolism and
in family members of those with a mutation: a systematic review. JAMA 2009, 301:2472-85.
7. Kujovich JL. Prothrombin Thrombophilia. 2006 Jul 25 [Updated 2021 Feb 4]. Accessed at
https://www.ncbi.nlm.nih.gov/books/NBK1148/.
8. Zhang S, Taylor AK, Huang X, et al. Venous thromboembolism laboratory testing (factor V Leiden and factor II c.*97G>A),
2018 update: a technical standard of the American College of Medical Genetics and Genomics (ACMG). Genet Med. 2018
Dec;20(12):1489-1498. Epub 2018 Oct PubMed 30297698

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Jagadeesh Babu S A. Radhika Dr. Naushad SM

Dr. Kiran Kumar

Technical Lead -Genomics Co-Head Genomics Chief Scientific Officer Consultant Genomics

6-3-862/A, Lal Bungalow, Ameerpet, Hyderabad – 500016, info@lifelinediagnostics.in,

Ph: +91 9542129919