Amplificatietesten

Amplificatietesten are diagnostic tests that detect biological targets by amplifying nucleic acids. The principle is to increase the quantity of a specific DNA or RNA sequence so that it can be detected even when present in very small amounts. The most common approaches are nucleic acid amplification tests (NAATs), including polymerase chain reaction (PCR)-based methods and isothermal amplification techniques.

PCR-based methods include conventional PCR, real-time quantitative PCR (qPCR), reverse transcription PCR (RT-PCR) for RNA targets, and digital PCR. Isothermal methods such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA) and helicase-dependent amplification operate at a constant temperature, enabling faster and often simpler workflows.

Applications span medical diagnostics, infectious disease surveillance, genetic and oncology testing, and environmental or food safety monitoring. In clinical practice, amplification tests are used to detect pathogens (for example SARS-CoV-2 and HIV), to identify genetic mutations, and to quantify nucleic acids for prognosis or treatment monitoring.

A typical workflow includes specimen collection, extraction of nucleic acids, the amplification step, and detection of amplified products through fluorescence, colorimetric signals, or lateral flow readouts. The method’s sensitivity and specificity depend on primer design, sample quality, and laboratory controls.

Advantages of amplification tests include high sensitivity, specificity, and the possibility of multiplexing. Limitations include the risk of contamination leading to false positives, the need for specialized equipment and trained personnel, potential inhibitors in samples, and higher costs compared to some non-amplification tests. Quality assurance, validation, controls, and accreditation are essential for reliable results.