How to Test

This is for US residents only.

Test overview1

 

The QIAGEN therascreen® PIK3CA RGQ PCR Kit is an FDA-approved companion diagnostic (CDx) test for the detection of 11 mutations in the PIK3CA gene using genomic DNA (gDNA) extracted from formalin-fixed, paraffin-embedded (FFPE) breast tumor tissue or circulating tumor DNA (ctDNA) isolated from K2EDTA anticoagulated peripheral whole blood plasma taken from patients with breast cancer.

Although a plasma test is FDA approved, it has not yet been verified by any US laboratory. Revisit this website for updates.

 

Limit of detection: tissue specimens1

 

A study was conducted to determine the limit of detection (LoD) of each of the 11 PIK3CA mutations. LoD was defined as the lowest amount of mutant DNA in a background of wild-type DNA at which a mutant sample will provide mutation-positive results in 95% of the test results (C95). The LoDs for the 11 PIK3CA mutation assays of the therascreen® PIK3CA Rotor-Gene Q (RGQ) Kit are reported as Mutant Allele Frequency (MAF).

 

 

Limit of detection for each of the 11 PIK3CA mutations1

 

 
Exon
Mutation
COSMIC* ID
Base change
LoD (% MAF)
 
 
7
9
20
 
C420R
E542K
E545A
E545D
E545G
E545K
Q546E
Q546R
H1047L
H1047R
H1047Y
 
757
760
12458
765
764
763
6147
12459
776
775
774
 
1258T>C
1624G>A
1634A>C
1635G>T
1634A>G
1633G>A
1636C>G
1637A>G
3140A>T
3140A>G
3139C>T
 
2.41
5.47
3.54
2.69
4.98
4.13
4.50
6.08
2.56
3.13
14.04
 
 

 

MAF, mutant allele frequency.
*COSMIC, Catalogue of Somatic Mutations in Cancer: https://cancer.sanger.ac.uk/cosmic.
LoD values were established using DNA from cell-line specimens.
LoD values were established using DNA from clinical specimens.

 

Limit of detection: plasma specimens1

A study was conducted to determine the LoD of each of the 11 PIK3CA mutations using contrived plasma specimens. LoD was defined as the lowest amount of mutant DNA in a background of wild-type DNA at which a mutant sample will provide mutation-positive results in 95% of the test results (C95).

 

Limit of detection for each of the 11 PIK3CA mutations1

 

 
Exon
Mutation
COSMIC* ID
Base change
LoD (% MAF)
 
 
7
9
20
 
C420R
E542K
E545A
E545D
E545G
E545K
Q546E
Q546R
H1047L
H1047R
H1047Y
 
757
760
12458
765
764
763
6147
12459
776
775
774
 
1258T>C
1624G>A
1634A>C
1635G>T
1634A>G
1633G>A
1636C>G
1637A>G
3140A>T
3140A>G
3139C>T
 
4.46
5.06†‡
1.82
3.21
1.94†‡
2.42†‡
5.31
4.22
2.37†‡
1.98†‡
7.07
 
 

 

MAF, mutant allele frequency.
*COSMIC: Catalogue of Somatic Mutations in Cancer: https://cancer.sanger.ac.uk/cosmic.
LoD values were established using cell-line specimens.
LoD values were verified using clinical plasma specimens.

Preparing the test sample1

The QIAGEN therascreen® PIK3CA RGQ PCR Kit is for use with gDNA extracted from FFPE tumor tissue resected specimens and core needle biopsy (CNB) specimens collected from breast cancer patients. Tumors are heterogeneous in terms of both genotype and phenotype. Mutation-positive tumors can contain wild-type DNA and histology can similarly show regions of non-tumor tissue.

 

 

Block for samples


OR

Slides for samples

There are two separate workflows when using FFPE tumor tissue resected specimens and FFPE CNB specimens.1

 

FFPE CNB specimen workflow

 

FFPE tumor tissue resected specimen workflow

 

 

The block or 6-12 slides may be sent. For more information, contact NeoGenomics at 1-800-755-1605. The block or 6-12 slides may be sent. For more information, contact NeoGenomics at 1-800-755-1605.

 

Label, handle, and store tumor specimens, blocks, slides, samples, and microcentrifuge tubes ready for extraction in a controlled fashion according to local procedures.

It is important to follow correct specimen storage conditions, reagent shipping conditions, and reagent storage conditions. For more information, please refer to the Instructions for Use.

 

 

Test results

When using the QIAGEN Rotor-Gene Q MDx (US) instrument, the following results may be assigned to an individual sample:

  • PIK3CA Mutation Detected

  • No Mutation Detected

  • INVALID: If one or more flags are assigned to the sample during analysis by the Rotor-Gene AssayManager v2.1 software are defined to set the target result to “INVALID”1

Note: If an error has occurred during the run, the samples in the Rotor-Gene Q MDx (US) must be disposed of and not be retested.

 

For detailed instructions regarding sample collection and preparation protocols, please refer to the Instructions for Use.

 

 

Indication

PIQRAY® (alpelisib) tablets is indicated in combination with fulvestrant for the treatment of postmenopausal women, and men, with hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative, PIK3CA-mutated, advanced or metastatic breast cancer as detected by an FDA-approved test following progression on or after an endocrine-based regimen.

Important Safety Information
PIQRAY is contraindicated in patients with severe hypersensitivity to it or any of its components.

Severe Hypersensitivity: Severe hypersensitivity reactions, including anaphylaxis and anaphylactic shock, were reported in patients treated with PIQRAY. Severe hypersensitivity reactions were manifested by symptoms including, but not limited to, dyspnea, flushing, rash, fever, or tachycardia. Advise patients of the signs and symptoms of severe hypersensitivity reactions. Permanently discontinue PIQRAY in the event of severe hypersensitivity.

Severe Cutaneous Reactions: Severe cutaneous reactions, including Stevens-Johnson syndrome (SJS) and erythema multiforme (EM) were reported. Do not initiate PIQRAY treatment in patients with a history of SJS, EM, or toxic epidermal necrolysis (TEN). Interrupt PIQRAY if signs or symptoms of severe cutaneous reactions are present (eg, a prodrome of fever, flu-like symptoms, mucosal lesions or progressive skin rash), until etiology of the reaction has been determined. Advise patients of the signs and symptoms of severe cutaneous reactions. Consider consultation with a dermatologist. Permanently discontinue PIQRAY if SJS, EM, or TEN is confirmed.

Hyperglycemia: Severe hyperglycemia, including ketoacidosis, was reported. Before initiating treatment with PIQRAY, test fasting plasma glucose (FPG), HbA1c, and optimize blood glucose. After initiating treatment, monitor blood glucose and/or FPG at least once every week for the first 2 weeks, then at least once every 4 weeks, and as clinically indicated. Monitor HbA1c every 3 months and as clinically indicated. Initiate or optimize antihyperglycemic medications as clinically indicated. Interrupt, reduce dose, or discontinue PIQRAY if severe hyperglycemia occurs. The safety of PIQRAY in patients with type 1 and uncontrolled type 2 diabetes has not been established as these patients were excluded from the SOLAR-1 trial. Patients with a history of diabetes mellitus may require intensified diabetic treatment. Closely monitor patients with diabetes.

Advise patients of the signs and symptoms of hyperglycemia (eg, excessive thirst, urinating more often than usual or higher amount of urine than usual, or increased appetite with weight loss).

Pneumonitis: Severe pneumonitis, including acute interstitial pneumonitis and interstitial lung disease, has been reported in patients treated with PIQRAY. Monitor for clinical symptoms or radiological changes. Consider a diagnosis of non-infectious pneumonitis in patients presenting with non-specific respiratory signs and symptoms such as hypoxia, cough, dyspnea, or interstitial infiltrates on radiologic exams and in whom infectious, neoplastic, and other causes have been excluded by means of appropriate investigations. Interrupt or discontinue PIQRAY if severe pneumonitis occurs. Advise patients to immediately report new or worsening respiratory symptoms.  

Diarrhea: Severe cases of diarrhea, including dehydration and acute kidney injury, have been reported. Based on the severity of the diarrhea, PIQRAY may require dose interruption, reduction, or discontinuation. Advise patients to start antidiarrheal treatment, increase oral fluids, and notify their health care provider if diarrhea occurs while taking PIQRAY.

Embryo-Fetal Toxicity: PIQRAY and fulvestrant can cause fetal harm. Advise patients of potential risk to a fetus and to use effective contraception. Also, refer to Full Prescribing Information of fulvestrant for pregnancy and contraception information.

Most common adverse reactions, including laboratory abnormalities (all grades, incidence ≥20%) were glucose increased (79%), creatinine increased (67%), diarrhea (58%), rash (52%), lymphocyte count decreased (52%), gamma glutamyl transferase increased (52%), nausea (45%), alanine aminotransferase increased (44%), fatigue (42%), hemoglobin decreased (42%), lipase increased (42%), decreased appetite (36%), stomatitis (30%), vomiting (27%), weight decreased (27%), calcium decreased (27%), glucose decreased (26%), activated partial thromboplastin time  prolonged (21%), and alopecia (20%).

Please click here for full Prescribing Information.

Reference: 1. therascreen® PIK3CA RGQ PCR Kit Instructions for Use. Germantown, MD: QIAGEN; May 2019.

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