• High precision and sensitivity: The use of duplex, hydrolysis probe-based assays allows for highly precise detection of mutations. The presence of both mutant and wild-type probes in the same reaction ensures that researchers can detect and quantify minor genetic variations with great accuracy, crucial for studies in heterogeneous cancer samples where only a few cells may carry the mutation.
• Enhanced specificity: The integration of LNA into the probes increases the binding affinity and specificity towards the target sequences, minimizing the risk of non-specific bindings and improving the overall reliability of the assays.
• Multiplexing capability: Each assay is capable of detecting mutations using two different fluorescent dye combinations, allowing for the simultaneous analysis of mutant and wild-type alleles within the same reaction. This multiplexing ability is particularly useful in applications requiring the analysis of multiple targets, such as assessing co-occurring mutations in cancer.
• Flexibility in sample analysis: By dividing the reaction across multiple wells, even greater sensitivity can be achieved, facilitating the detection of extremely rare mutations. This is especially valuable in cancer research, where detecting low-frequency mutations can inform prognosis and treatment strategies.
• Streamlined workflow: Supplied in a single-tube format with ready-to-use primer pairs and probes, these assays simplify the experimental setup, enabling efficient and straightforward integration into existing research workflows.

• AR c.2632A>G / p.T878A: This point mutation results in a threonine-to-alanine substitution at position 878 that alters the ligand-binding domain of the AR, allowing it to be activated by other steroids and anti-androgens. This broadened ligand specificity disrupts normal regulatory mechanisms, enabling AR to drive gene expression that supports cancer cell survival and proliferation. The T878A mutation has also been associated with increased resistance to current anti-androgen therapies, necessitating the development of next-generation inhibitors to effectively target this variant.
• AR c.2623C>T / p.H875Y: This point mutation results in a histidine-to-tyrosine substitution at position 875 that also affects the ligand-binding domain and alters receptor activation. The H875Y variant increases the receptor's affinity for alternative ligands, which can lead to aberrant activation of AR signaling pathways. This change facilitates cancer progression by promoting uncontrolled cell division and resistance to apoptosis. The altered receptor dynamics in the presence of the H875Y mutation pose significant challenges for the efficacy of traditional androgen deprivation therapies, highlighting the need for more personalized treatment approaches.

• BRCA2 c.7234_7235insG: This variant is an insertion mutation that results in a frameshift, leading to a premature stop codon. This mutation disrupts the normal function of the BRCA2 protein, which is essential for homologous recombination repair of DNA double-strand breaks. The loss of functional BRCA2 protein leads to genomic instability, promoting tumorigenesis and progression of prostate cancer. This variant has been identified in Moroccan prostate cancer patients with a positive family history, indicating its significance in hereditary prostate cancer.
• BRCA2 ΔE12: This variant involves a deletion of exon 12, which results in the loss of a significant portion of the BRCA2 protein. This deletion impairs the protein's ability to interact with RAD51, a key protein in the homologous recombination repair pathway. The resulting deficiency in DNA repair mechanisms leads to increased susceptibility to DNA damage and contributes to the development and progression of prostate cancer. This variant has also been reported in Moroccan patients, highlighting its role in familial prostate cancer.
• BRCA2 c.1280_1281delGA / p.Asp427Thrfs*3: This variant is a frameshift mutation that introduces a premature stop codon, truncating the BRCA2 protein. This truncation eliminates critical domains required for DNA repair, leading to defective homologous recombination and increased genomic instability. This variant has been identified in Japanese patients with castration-resistant prostate cancer, suggesting its relevance in advanced stages of the disease.

• CHEK2 c.1100delC: This variant is a frameshift mutation that results in a truncated CHEK2 protein, impairing its function in DNA repair and cell cycle control. This mutation is associated with an increased risk of several cancers, including prostate cancer. The loss of CHEK2 function compromises the cell's ability to respond to DNA damage, contributing to genomic instability and cancer development.
• CHEK2 c.470T>C / p.I157T: The I157T variant is a missense mutation that alters the protein structure and function of CHEK2, leading to a compromised DNA damage response. This variant is associated with an increased risk of prostate cancer and other cancers, such as breast and thyroid cancer. The altered protein function due to this mutation diminishes the cell's ability to effectively repair DNA, increasing the likelihood of cancerous transformations.

The KRAS gene is a proto-oncogene that encodes a small GTPase involved in the RAS/MAPK signaling pathway. This pathway regulates cell proliferation, differentiation and survival. Mutations in KRAS lead to constitutive activation of the RAS/MAPK pathway, driving oncogenic processes. Although KRAS mutations are less common in prostate cancer compared to other cancers, they are associated with more aggressive disease and poor prognosis when present. The constitutive activation of KRAS due to mutations enhances oncogenic signaling, contributing to tumor aggressiveness and resistance to treatment.

• PTEN c.388C>T / p.Arg130*: This variant in the PTEN gene results in a nonsense mutation, leading to a truncated protein that lacks functional phosphatase activity. This mutation disrupts the normal regulation of the PI3K/AKT pathway, promoting tumorigenesis and progression of prostate cancer. The loss of PTEN function due to this mutation allows for unregulated cell growth and survival, contributing to more aggressive cancer phenotypes.
• PTEN c.697C>T / p.Arg233*: This variant is another nonsense mutation that results in a truncated PTEN protein. This mutation impairs the tumor suppressor function of PTEN, contributing to the development and progression of prostate cancer. By disrupting the PI3K/AKT pathway regulation, this mutation leads to enhanced cellular proliferation and resistance to apoptosis, exacerbating the severity of the disease.

• SPOP c.397T>G / p.F133V: The F133V variant results in a phenylalanine-to-valine substitution at position 133. This mutation affects the substrate-binding cleft of the SPOP protein, altering its ability to target specific proteins for degradation. The F133V mutation is associated with increased sensitivity to androgen deprivation therapy (ADT) in prostate cancer. By changing the substrate affinity, this mutation can influence the degradation rates of critical regulatory proteins, impacting tumor growth.
• SPOP c.260A>G / p.Y87C: The Y87C variant involves a tyrosine-to-cysteine substitution at position 87. This mutation also affects the substrate-binding cleft, leading to dysregulation of key regulatory pathways in prostate cancer. The Y87C mutation is linked to increased tumor growth and progression. The altered binding dynamics due to this mutation result in the improper degradation of proteins that regulate cell proliferation and survival.

• TP53 c.818G>A / p.R273H: This variant results in an arginine-to-histidine substitution at position 273. This mutation affects the DNA-binding domain of p53, impairing its ability to regulate target genes involved in cell cycle control and apoptosis. The R273H mutation is associated with poor prognosis and resistance to therapy in prostate cancer. By compromising the DNA-binding capability, this mutation allows for unchecked cell proliferation and survival.
• TP53 c.524G>A / p.R175H: This variant involves an arginine-to-histidine substitution at position 175. This mutation disrupts the structural integrity of the p53 protein, leading to loss of its tumor suppressor function. The R175H mutation is commonly found in aggressive forms of prostate cancer. This structural alteration prevents p53 from effectively controlling the cell cycle and promoting apoptosis in damaged cells.