Porównanie trzech testów multiplex real-time PCR do wykrywania DNA wirusów opryszczki typu 1 i 2
[Comparison of three multiplex real-time PCR assays for the detection of herpes simplex viruses type 1 and type 2 DNA]

Ludzkie wirusy opryszczki typu 1 i typu 2 (HSV-1 i HSV-2) wywołują powszechne na całym świecie zakażenia o różnorodnym przebiegu i nasileniu. Celem pracy było porównanie diagnostycznej wartości zaprojektowanych w Katedrze i Zakładzie Mikrobiologii Lekarskiej WUM dwóch metod multiplex realtime PCR z użyciem sond typu TaqMan® oraz HybProbes® do wykrywania DNA HSV-1 i HSV-2 z komercyjną metodą opartą na układzie starterów/sond

SCORPIONS™.

ABSTRACT

Introduction: Herpes simplex viruses type 1 and type 2 (HSV-1 and HSV-2) causewidespread infection worldwide with different course and intensity. Although the disease caused by this viruses mainly concern healthy children and adults, the HSV infections are much more dangerous for people with immunodeficiencies. The aim of this work was to compare the diagnostic value of two qPCR methods for detection HSV-1/2 DNA, based on TaqMan® and HybProbes® chemistry with commercial HSV-1/2 Qual Kit.

Methods: DNA from 51 clinical samples was tested for presence of HSV-1/2 sequences on LightCycler 2.0 thermocycler, using two „in-house” developed multiplex real-time PCR assays and commercial test using SCORPIONS™ primers.

Results: The results showed high specificity and sensitivity of all molecular biology tests used. Statistically, there was no significant difference in the sensitivity of real-time PCR assays when using TaqMan® and HybProbes® chemistry and when using the commercial SCORPIONS™ based method (P>0.05).

Conclusions: Obtained results show that all tested methods are highly specific and can possibly be used to simultaneously detect and differentiate HSV-1/2 DNA in clinical samples. The high detection rate and short duration of qPCR assays has great importance for immunocompromised patients where quick application of effective and safe treatment is necessary. It is also important in the event of amorphous form of the infection and the occurrence of nonspecific and generalized symptoms.