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 Video

Updated: May 28, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping
13:06

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

A PIK3CA pyrosequencing-based assay that excludes pseudogene interference.

Christine L Baker1, Cecily P Vaughn, Wade S Samowitz

  • 1ARUP Institute for Clinical and Experimental Pathology, University of Utah Health Sciences Center, Salt Lake City, Utah 84108, USA.

The Journal of Molecular Diagnostics : JMD
|October 27, 2011
PubMed
Summary
This summary is machine-generated.

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

Empirical estimate for the indirect healthcare costs of sickle cell disease from a retrospective cohort study in England between 2007 and 2019.

BMC health services research·2026
Same author

Economic Burden of Sickle Cell Disease: A Retrospective Study of Pediatric and Adult Individuals With Medicaid Coverage From 2016 to 2020.

American journal of hematology·2025
Same author

Economic burden of sickle cell disease in the United States: a retrospective analysis of a commercial insurance database.

Journal of comparative effectiveness research·2025
Same author

Clinical and economic burden of achondroplasia in the United States: results from a retrospective, observational study.

Orphanet journal of rare diseases·2025
Same author

Development of a Conceptual Model for the Patient Experience of Immunoglobulin A Nephropathy (IgAN): A Qualitative Literature Review.

Advances in therapy·2024
Same author

Development of a Conceptual Model for the Patient Experience of Focal Segmental Glomerulosclerosis (FSGS): A Qualitative Targeted Literature Review.

Advances in therapy·2023
Same journal

Epigenetic CD4+ T-Cell Quantification from Dried Blood Spots Using a qPCR-Based Assay.

The Journal of molecular diagnostics : JMD·2026
Same journal

Segmental Copy Number Variant Detection Using an Amplicon-based NGS Panel for Integrated Glioma Classification.

The Journal of molecular diagnostics : JMD·2026
Same journal

Clinical Validation of the Roche cobas and cobas 4800 Human Papillomavirus Tests on Self-Collected Vaginal Dry Swabs versus Practitioner-Collected Cervical Specimens Using the VALHUDES Protocol.

The Journal of molecular diagnostics : JMD·2026
Same journal

Long-Read Nanopore Sequencing Enhances BRCA1/2 Variant Detection Compared with Ion Torrent Analysis.

The Journal of molecular diagnostics : JMD·2026
Same journal

Endonuclease-Assisted Selective Exponential Amplification for Ultrasensitive Enrichment and Detection of Low-Abundance Mutant Alleles in Lung Cancer.

The Journal of molecular diagnostics : JMD·2026
Same journal

Validation of NTRK Fusion Detection Using an Ultrarapid, Fully Automated Cartridge-Based PCR Assay.

The Journal of molecular diagnostics : JMD·2026
See all related articles

A new pyrosequencing assay accurately detects Phosphatidylinositol 3'-kinase gene (PIK3CA) mutations in colorectal cancer, crucial for targeted therapy. This method avoids pseudogene interference, improving diagnostic reliability for PIK3CA in cancer research.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Phosphatidylinositol 3 -kinase gene (PIK3CA) mutations are prevalent in colorectal cancer (CRC), affecting the epidermal growth factor receptor pathway.
  • Activating PIK3CA mutations upregulate AKT signaling, presenting a potential therapeutic target in CRC.
  • PIK3CA mutations can co-occur with KRAS, BRAF, or NRAS mutations in CRC.

Purpose of the Study:

  • To develop and validate a pyrosequencing assay for detecting hotspot mutations in PIK3CA.
  • To ensure the assay avoids amplification of a homologous PIK3CA pseudogene.
  • To evaluate the prevalence of PIK3CA mutations in a cohort of colorectal cancer patients.

Main Methods:

  • Design of a pyrosequencing assay targeting codons 542, 545 (exon 9), and 1047 (exon 20) of PIK3CA.

More Related Videos

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
06:53

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
07:24

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Related Experiment Videos

Last Updated: May 28, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping
13:06

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
06:53

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
07:24

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

  • Development of a specific exon 9 reverse PCR primer to prevent pseudogene amplification.
  • Analysis of PIK3CA mutations in 200 colorectal cancer samples from FFPE tissues, with prior KRAS mutation status.

    • Main Results:

    • The assay successfully detected PIK3CA mutations in 20% of KRAS-mutated CRC samples and 7.5% of KRAS wild-type samples.
    • No pseudogene amplification was observed in any of the analyzed samples.
    • Validation studies confirmed gene-specific and pseudogene-specific amplification, demonstrating assay specificity.

    Conclusions:

    • A robust PIK3CA pyrosequencing assay has been developed, capable of detecting mutations in key hotspot codons.
    • The assay effectively differentiates between the PIK3CA gene and its pseudogene, ensuring accurate mutation detection.
    • This validated assay can aid in identifying therapeutic targets in colorectal cancer patients.