Reverse Transcription Polymerase Chain Reaction (RT-PCR)
Reverse transcription polymerase chain reaction (RT-PCR) is a molecular biology technique used to amplify and detect specific RNA sequences, including viral RNA, in biological samples. Here’s how it’s used:
- Diagnosis of Infectious Diseases: RT-PCR is widely used for the diagnosis of infectious diseases caused by RNA viruses, including respiratory viruses (e.g., influenza virus, respiratory syncytial virus, SARS-CoV-2), bloodborne viruses (e.g., HIV, hepatitis C virus), and other viral pathogens (e.g., dengue virus, Zika virus). By detecting viral RNA in clinical specimens such as nasal swabs, throat swabs, blood, or cerebrospinal fluid, RT-PCR helps confirm the presence of viral infection.
- Screening and Surveillance for Pathogens: RT-PCR is used for population-based screening and surveillance programs to monitor the prevalence and transmission of infectious diseases within communities. Large-scale RT-PCR testing, particularly during outbreaks or pandemics, provides valuable epidemiological data for public health authorities to implement control measures and allocate resources effectively.
- Detection of Genetic Mutations and Variants: RT-PCR can be used to detect specific genetic mutations or variants of pathogens, such as those associated with drug resistance, immune evasion, or increased transmissibility. Monitoring the prevalence of genetic variants helps track the evolution of pathogens and informs public health strategies, such as vaccine development and treatment protocols.
- Quantification of Gene Expression: RT-PCR can quantify gene expression levels by measuring the amount of RNA transcripts present in biological samples. This quantitative RT-PCR (qRT-PCR) technique is commonly used in research and clinical laboratories to study gene expression patterns, identify biomarkers, and assess disease progression in various conditions, including cancer, autoimmune diseases, and neurological disorders.
- Detection of Gene Fusions and Aberrations: RT-PCR can detect specific gene fusions, rearrangements, or aberrations associated with genetic diseases, such as leukemia, lymphoma, or solid tumors. By amplifying and analyzing target gene sequences, RT-PCR helps diagnose genetic disorders, classify disease subtypes, and guide treatment decisions in precision medicine approaches.