Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Two rare confounding polymorphisms proximal to the factor V Leiden mutation.

B Gold1, M Hanson, M Dean

  • 1Quest Diagnostics, Van Nuys, CA, USA. goldb@ncifcrf.gov

Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology
|July 27, 2001
PubMed
Summary

Novel intron polymorphisms can interfere with Factor V Leiden (G1691A) genotyping using PCR-RFLP. Sequencing identified two new polymorphisms in intron 10, resolving unusual restriction patterns in clinical diagnostic tests.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Estimating the prevalence of active Helicobacter pylori infection in a rural community with global positioning system technology-assisted sampling.

Epidemiology and infection·2012
Same author

Chameleonic reactivity of vicinal diazonium salt of acetylenyl-9,10-anthraquinones: synthetic application toward two heterocyclic targets.

The Journal of organic chemistry·2011
Same author

Generalized schemes for high-throughput manipulation of the Desulfovibrio vulgaris genome.

Applied and environmental microbiology·2011
Same author

The E. coli monothiol glutaredoxin GrxD forms homodimeric and heterodimeric FeS cluster containing complexes.

Biochemistry·2011
Same author

Pharmacological inhibition of poly(ADP-ribose) polymerase (PARP) activity in PARP-1 silenced tumour cells increases chemosensitivity to temozolomide and to a N3-adenine selective methylating agent.

Current cancer drug targets·2010
Same author

Genetic variants in AVPR1A linked to autism predict amygdala activation and personality traits in healthy humans.

Molecular psychiatry·2008

Area of Science:

  • Molecular diagnostics
  • Genetic testing
  • Thrombophilia risk assessment

Background:

  • Factor V Leiden mutation (G1691A) is crucial for thrombophilia risk assessment.
  • PCR-restriction fragment length polymorphism (PCR-RFLP) is a common genotyping method.
  • Primer annealing within intron 10 can lead to confounding results due to polymorphisms.

Purpose of the Study:

  • To investigate unusual electrophoretic mobilities observed during Factor V G1691A genotyping.
  • To identify novel polymorphisms that may confound PCR-RFLP results.

Main Methods:

  • Analysis of 15,301 patient samples undergoing thrombophilia risk assessment.
  • PCR amplification and restriction enzyme digestion.
  • DNA sequencing of amplicons with unusual restriction patterns.

Related Experiment Videos

Main Results:

  • Two novel polymorphisms were identified within intron 10 of the Factor V gene.
  • These polymorphisms explained unusual electrophoretic patterns in two patient samples.
  • Sequencing confirmed the presence of these novel variants.

Conclusions:

  • PCR-RFLP methods are susceptible to confounding by polymorphisms near primer binding sites, especially within introns.
  • Novel intron polymorphisms can mimic or mask true mutations, impacting diagnostic accuracy.
  • DNA sequencing is a valuable tool for resolving ambiguous PCR-RFLP results and identifying new genetic variants.