PIK3CA Mutation Analysis, therascreen - QIAGEN (FDA approved)

CPT 0155U; 88381
Synonyms
  • PIK3CA

Test Details

Cancer Type

Breast

Technology Used

Molecular

Turnaround Time

5 - 7 days

Use

The therascreen PIK3CA RGQ RT-PCR Kit is a real-time, qualitative PCR assay for the detection of 11 mutations in the phosphatidyl 3-kinases catalytic subunit alpha (PIK3CA) gene (Exon 7: C420R; Exon 9: E542K; E545A, E545D [1635G>T only], E545G, E545K, Q546E, Q546R; and Exon 20: H1047L, H1047R, H1047Y) using genomic DNA (gDNA) extracted from formalin-fixed paraffin-embedded (FFPE) breast tumor tissue. The test is intended to aid clinicians in identifying breast cancer patients who may be eligible for treatment with PIQRAY(R) (alpelisib) based on a PIK3CA Mutation Detected result. Patients whose FFPE tissue produce a positive therascreen PIK3CA RGQ PCR Kit test result for the presence of one or more PIK3CA mutations are eligible for treatment with PIQRAY® (alpelisib).

Table 1. List of mutations detected and COSMIC identities

Exon

Mutation

COSMIC* ID

Base Change

7

C420R

757

1258 T>C

9

E542K

760

1624 G>A

9

E545A

12458

1634 A>C

9

E545D

765

1635G>T

9

E545G

764

1634A>G

9

E545K

763

1633 G>A

9

Q546E

6147

1636 A>G

9

Q546R

12459

1637 A>G

20

H1047L

776

3140 A>T

20

H1047R

775

3140 A>G

20

H1047Y

774

3139 C>T

* COSMIC: Catalogue of somatic mutations in cancer: https://cancer.sanger.ac.uk/cosmic

Special Instructions

Please provide a copy of the pathology report. Testing will be delayed if the pathology report is not received. Please direct any questions regarding this test to customer service at 800-345-4363.

Limitations

Results from the product must be interpreted within the context of all relevant clinical and laboratory findings and are not to be used alone for diagnosis.

Samples with results reported as "No Mutation Detected" may harbor PIK3CA mutations not detected by the therascreen PIK3CA RGQ PCR Kit.

Detection of mutations is dependent on sample integrity and the amount of amplifiable DNA present. The test procedure should be repeated if analysis of the DNA in the sample indicates that the quantity and/or quality is either not sufficient or the concentration is too high for mutation analysis.

The therascreen PIK3CA RGQ PCR Kit is used in a PCR procedure. As with all PCR procedures, samples may become contaminated by external sources of DNA in the test environment and the DNA in the positive control. Use caution to avoid contamination of samples and kit reagents.

If the sample contains less than the percentage of mutant alleles that is able to be detected by the therascreen PIK3CA RGQ PCR Kit, it will lead to a result of "No Nutation Detected."

It is not known whether the therascreen PIK3CA RGQ PCR Kit shows cross-reactivity (results of "Mutation Detected") to additional PIK3CA mutations different than those included in the kit.

The therascreen PIK3CA RGQ PCR Kit is a qualitative test. The test will not provide quantitative measurements of the Mutant Allele Frequency (MAF) present in a sample.

The impact of the performance of the therascreen PIK3CA RGQ PCR Kit is unknown if microbial contamination is introduced during assay procedures; operators must exercise due caution to avoid introduction of microbial contaminants during testing procedures and should not use kit components if evidence of microbial growth is observed.

The therascreen PIK3CA RGQ PCR Kit is only for use with DNA extracted from formalin-fixed, paraffin-embedded breast cancer tissue.

The therascreen PIK3CA RGQ PCR Kit is only for use with the QIAamp DSP DNA FFPE Tissue Kit (for tissue specimens).

Methodology

The therascreen PIK3CA RGQ PCR Kit is a real-time qualitative PCR in vitro diagnostic test, performed on the Rotor-Gene Q MDx (US) instrument. It uses allele refractory mutation system (ARMS) primers, hydrolysis probes and PCR clamp technologies to detect 11 mutations in exons 7, 9 and 20 of the PIK3CA oncogene against a background of wild-type gDNA.

Specimen Requirements

Information on collection, storage, and volume

Specimen

Formalin-fixed, paraffin-embedded tissue (FFPE) block or slides from tissue resection or core needle biopsy (CNB)

Volume

Formalin-fixed, paraffin-embedded tissue (FFPE) block or 6 unstained slides at 4-5m and one matching H&E-stained slide or 7 unstained slides

Minimum Volume

3 unstained slides at 4-5m and one matching H&E-stained slide or 4 unstained slides. Resection or Surgical Biopsies require > or =10% tumor content. Core Needle Biopsies require a minimum of > or = 20 mm2 tumor area total available for extraction. Minimum volume allows only one attempt at DNA extraction.

Container

FFPE block or slides

Storage Instructions

Maintain blocks/slides at room temperature.

Causes for Rejection

Specimen does not meet all of the above criteria for sample type, container, minimum volume, collection and storage; specimens containing suspicious foreign material; no tumor tissue in FFPE block or slides; broken or stained slides; fixative other than formalin

Collection

Ship specimen at room temperature. Please direct any questions regarding this test to customer service at 800-345-4363.