The CYP3A family is the drug metabolizing enzyme that mediates the metabolism of the largest number of drugs in the human body. More than 50% of the oxidation and reduction reactions of commonly used clinical drugs are catalyzed by CYP3A4 and CYP3A5. CYP3A5 is involved in the metabolism of tacrolimus, midazolam, tranylcypromine, cortisone, nifedipine and many other drugs. A 6986A>G mutation (rs776746, CYP3A5*3) exists in intron 3 of the CYP3A5 gene at position 22893. The SNP could cause abnormal splicing of CYP3A5 mRNA, causing premature shearing of the CYP3A5 protein by the stop codon, thereby depriving it of enzymatic activity. As a result, homozygote CYP3A5*3 individuals had significantly reduced hepatic and intestinal CYP3A5 protein expression and activity.

Tacrolimus (FK506), a macrolide immunosuppressant, is widely used clinically for the immunosuppressive treatment of liver, kidney, heart, lung, and pancreas transplant patients. Its major adverse effects include secondary infections, nephrotoxicity, neurotoxicity, gastrointestinal reactions, metabolic disorders and lymphoproliferative disorders and tumors, etc. Low blood levels after tacrolimus administration in organ transplant patients can lead to acute rejection and reduced drug sensitivity. High blood levels predispose to adverse effects such as nephrotoxicity, neurotoxicity, diabetes, hyper lipidaemia, hypertension, and gastrointestinal disturbances.

This kit is only used for the detection of specific target gene sequences, and its detection results are only for clinical reference and should not be used as the only basis for individual treatment of patients. Clinicians should comprehensively judge the test results according to the patient’s condition, drug indications, treatment response and other laboratory detection indexes.

UltraDx CYP3A5 Gene Polymorphism Detection Kit uses ARMS-qPCR technology to design wild-type and mutant ARMS primers for human CYP3A5 gene. Combined with Taqman probe, the specific amplification products are detected. Only when the allele-specific primer 3’terminal base was completely complementary to the corresponding allele genotype template, the PCR amplification reaction can be performed normally to obtain a specific product while generating a fluorescent signal. The loci to be tested were judged by the fluorescence signal results of the wild / mutant system, and the genotype was determined by the relative Ct value.

The internal standard in the kit uses house-keeping gene primers combined with specific probes to monitor the detection process in the fully closed reaction system and can effectively monitor the occurrence of false negative. In order to avoid the interaction between fluorescent signals, polymorphism detection and housekeeping genes use fluorescent groups with different emission wavelengths, in which CYP3A5 gene polymorphism detection uses FAM fluorescent group; house-keeping gene uses Texas Red fluorescent group.

• Strong Specificity: Non-cross reactivity.
• Contamination control: dUTP/UNG systems to avoid carry over contamination of PCR products in subsequent amplification reactions.
• High sensitivity: 10ng/reaction.
• Internal control system: Prevent false negative results.

Applicable sample type: EDTA whole blood.

Applied Biosystems™ Real time PCR system 7500, ABI QuantStudio™5 Real-time PCR system, LightCycler® 480 PCR system, Bio-Rad CFX96 real-time PCR instrument. Ultrassay eQ9600 Real Time qPCR System etc.