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Zastosowanie metody real-time PCR i systemu LightCycler do wykrywania DNA wirusa zakaznego ronienia klaczy [EHV-1]

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Dzieciatkowski T. , Przybylski M. , Chmielewska A. , Tucholska A. , Turowska A. , Banbura M.

tom 64

nr 07

918-921

Equine herpesvirus type 1 (EHV-1) is one of the major horse diseases, causing considerable worldwide losses. A variety of techniques, including nested PCR, have been used to diagnose EHV-1 infections. In this paper, a real-time PCR assay that uses non-specific SYBR Green I® fluorochrome for the detection of EHV-1 DNA is described. This method does not require post-amplification manipulations, thereby reducing the risk of cross-contamination. The assay was sensitive enough to detect EHV-1 sequences in neuronal cell cultures and also different clinical samples. The technique is specific: it was not reactive with other herpesviruses or opportunistic bacterial pathogens such as Escherichia coli, Staphylococcus epidermidis and Enterococcus faecium. In comparison to virus isolation or the nested PCR used previously, the test was more sensitive and should be useful for the common diagnosis based on its specificity and rapidity.