DNA-PCR

This laboratory method is available from 01.02.2016!

PCR (Polymerase Chain Reaction) is a molecular biological method for the direct detection of pathogen nucleic acids (DNA/RNA) in a sample. The detectability of pathogen DNA/RNA in blood or tissue can be equated with the (temporary) presence of the pathogen itself. A positive PCR result thus provides direct proof of the presence of an infection with the pathogen detected. However, it should be noted that the presence of the pathogens in a sample and thus the detectability of specific pathogen nucleic acids depend on the corresponding pathogenicity mechanism and vary with the stage of the disease. PCR-based pathogen detection in the blood is therefore only useful in the acute stage of the disease, during reactivation or in cases of doubt. In the chronic stage, detection from tissue/biopsat/pointate is particularly advisable.

Before the actual PCR test, the total DNA/RNA is first isolated from the patient’s blood or tissue. These nucleic acids are then used as starting material in the desired pathogen-specific PCR test. In the PCR reaction, specific oligonucleotides (primers) are used for each pathogen to be detected, which specifically bind to only one specific gene of the pathogen and enable the multiplication and subsequent detection of the pathogen. Thus, a high specificity of the detection is achieved. In addition to the conventional PCR method, there are various further developments, such as the nested PCR, the real-time PCR or the qPCR variant, which achieve the highest sensitivity for pathogen detection.

For the detection of borreliosis pathogens, a proprietary in-house PCR test system was developed, which is based on several successive PCR reactions and detects different pathogens specifically and highly sensitively. BCA-lab is one of the few laboratories that is able to detect Borrelia miyamotoi. The molecular biological pathogen detection spectrum is complemented by a PCR-based Bartonella assay.

 

Material for blood analysis: 4 x large EDTA tubes

Material for tissue/biopsate analysis: 

1 – 3 pieces (>1 1mm³) in sterile vessel with 0.9 % NaCl solution at 4 °C; for longer (>24 hours) transport time freeze tissue without liquid in sterile vessel directly at -20 °C or -80 °C.

Analytical examination duration:

3 days

(Note: Taking into account the pre- and post-analytical work processes, the report of findings is usually prepared in approx. 2 weeks).

Accounting according to GOÄ:

  • 1 x digit 4780 Factor: 1.15
  • 1 x digit 4783 Factor: 1.15
  • 1 x digit 4785 Factor: 1.15

Further literature:

  • Eshoo, M. W., Protector, S. E., Crowder, C. D., Carolan, H. E. & Ecker, D. J. Achieving molecular diagnostics for Lyme disease. Expert Rev. mol. Diagn. 13, 875-83 (2013).
  • Eshoo, M. W. et al. Direct molecular detection and genotyping of Borrelia burgdorferi from whole blood of patients with early Lyme disease. PLoS One 7, 3-8 (2012).
  • Chan, K., Marras, S. a E. & Parveen, N. Sensitive multiplex PCR assay to differentiate Lyme spirochetes and emerging pathogens Anaplasma phagocytophilum and Babesia microti. BMC Microbiol. 13, 295 (2013).
  • Brettschneider, S., Bruckbauer, H., Klugbauer, N. & Hofmann, H. Diagnostic value of PCR for detection of Borrelia burgdorferi in skin biopsy and urine samples from patients with skin borreliosis. J. Clin. Microbiol. 36, 2658-65 (1998).
  • Cerar, T., Ružić-Sabljić, E., Glinšek, U., Zore, a. & Strle, F. Comparison of PCR methods and culture for the detection of Borrelia spp. in patients with erythema migrans. Clin. Microbiol. Infect. 14, 653–658 (2008).

 

Laboratory order