Molecular Genetic Tacrolimus CYP3A5 Gene Polymorphism - RT-PCR
Test Code
MOG58
Also Known As
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Test Parameters Included
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Department
MOLECULAR GENETICS
Methodology
RT-PCR (Real-Time PCR or Quantitative PCR for Genotyping)
For gene polymorphism analysis (i.e., genotyping specific variants), the method typically refers to Real-Time PCR (qPCR) or other PCR-based genotyping assays. While the term "RT-PCR" can sometimes refer to Reverse Transcription PCR, in the context of DNA polymorphism analysis, it generally implies a quantitative PCR approach for detecting specific single nucleotide polymorphisms (SNPs). The underlying principle involves:
* DNA Extraction: Genomic DNA is extracted from the patient's whole blood sample.
* PCR Amplification: Specific primers are used to amplify the regions of the CYP3A5 gene where key polymorphisms (e.g., *3, *6, *7) are located. These polymorphisms are single nucleotide changes.
* Real-Time Detection/Genotyping: In Real-Time PCR, fluorescent probes are typically used. These probes are designed to bind specifically to either the "wild-type" (normal) allele or the "mutant" (variant) allele. The fluorescent signal generated during PCR amplification allows for the real-time detection and quantification of each allele. This helps in determining the patient's genotype (e.g., *1/*1, *1/*3, *3/*3, etc.).
* Interpretation of Genotype to Phenotype: Based on the detected genotype, the patient's metabolizer phenotype for drugs metabolized by CYP3A5 can be inferred (e.g., extensive/normal, intermediate, or poor metabolizer).
This highly sensitive and specific method allows for the identification of genetic variations that influence an individual's ability to metabolize tacrolimus and other CYP3A5 substrates.
Sample Required
Whole Blood (WB) in EDTA
* Typically, 2-5 mL of Whole Blood collected in an EDTA (lavender-top) vacutainer tube is required. EDTA is the preferred anticoagulant as it preserves cellular DNA integrity.
* Proper collection and timely transport (preferably refrigerated) are crucial to ensure DNA quality for accurate analysis.
Preparation
Proper collection and handling of the blood sample are essential for accurate and reliable genetic testing. * No specific patient preparation is required (e.g., fasting or dietary restrictions). * Sam... Read more
Schedule Report
3 Days
Emergency Report
Yes
Frequently Asked Questions
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Test Description
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