REAL-TIME PCR TEST METHOD

An accurate and quick diagnosis of viral infections in infectious diseases allows you to decide on the appropriate treatment method. Such possibilities are provided by research based on molecular methods, in particular Real-Time PCR. At genXone, we perform all diagnostic tests based on this technique.

What is Real-Time PCR?

The Real-Time PCR method allows us to show the direct presence of genetic material of bacteria, viruses, fungi and parasites in the tested material. To make it possible, in the first step it is necessary to duplicate unique DNA fragments characteristic of specific pathogens, i.e. their amplification. In the case of detection of RNA viruses, an additional reverse transcription step is performed, which in the described method takes place simultaneously with the amplification. The analysis of the course of the reaction and the reading of the signal is possible thanks to the use of special fluorescently labelled molecular probes. The data that the instrument records in the form of the fluorescence level for individual dyes are automatically converted into a result by specialized software.

Real-time PCR allows you to monitor the course of the reaction already during the analysis, unlike the classic PCR reaction, where only the final product can be analyzed (after the amplification is completed).

Advantages of Real-Time PCR

Comparing Real-Time PCR to the classical PCR reaction or other methods of infection diagnostics, such as cell culture, one can notice the great advantage of this technique. Its distinctive features are:

  • High specificity and sensitivity, much higher than in conventional laboratory methods – when the test sample has a low viral load or the period of its excretion is short, then the concentration of the virus may drop rapidly during the first few days of infection, as a consequence the virus may be undetectable by other methods than Real-Time PCR.
  • Short waiting time for obtaining the result – the quick time of performing the analysis undoubtedly also affects the economic aspects. Namely, it shortens the patient’s stay in the hospital, lowers the costs of treatment and hospitalization, and prevents the spread of infections in society and the hospital environment.
  • Identification of multiple pathogens in one analysis – after obtaining a positive result for a single pathogen, further diagnostics of other viruses or bacteria are often not continued, but numerous studies have shown that respiratory tract infections caused by three or more viruses may constitute up to 20–50% of cases. Thanks to Real-Time PCR, we can identify several to several dozen pathogens at one time, which also allows for the detection of co-infections.
  • Identification of troublesome bacteria and atypical microorganisms – microbial cultivation of some bacteria is difficult or even impossible to carry out, often also very long. Therefore, molecular tests are the most effective method of detecting pathogens in the active phase of the disease.
  • Ease of obtaining material for research – in the case of gastrointestinal infections, the tested material is faeces, and for urogenital infections – a swab.
  • A small amount of material is needed to perform the test – due to the high sensitivity of Real-Time PCR, the analysis can be performed on the basis of a small amount of collected material.
  • Minimal risk of making a mistake – this method is fully automated. The stage of isolation and assembly of reactions is performed by a robot, and the results are analyzed by specialized software. Thanks to this, there is practically no risk of making a mistake both at the stage of execution and interpretation of the results.

What infections are detected?

The Real-Time PCR method enables the detection of bacterial, viral and fungal infections as well as any parasitic infections. The genXone offer includes tests that allow detecting genetic material of pathogens causing infections of the digestive, respiratory and genitourinary systems as well as meningitis and encephalitis.