Tomato black ring virus (TBRV) infects a wide range of economically important plants, ornamental species and trees. The population of the virus is very diverse and the concentration of particles in plants depends on many factors such as isolate, host and climatic conditions. This causes serious problems in the development of effective and rapid methods to detect genetically diverse isolates. The methods used for the direct detection of the virus in the infected plant material, e.g. immunosorbent assays, do not always give satisfactory results. An immunocapture reverse transcription loop-mediated isothermal amplification (IC-RT-LAMP) and immunocapture real-time reverse transcription polymerase chain reaction (IC-RT-qPCR) protocols were developed for the detection of TBRV in different hosts. Eight isolates from black locus (Robinia pseudoacacia L.), one from tomato (Solanum lycopersicum cultivar Moneymaker) and one from zucchini (Cucurbita pepo convar. giromontiina cultivar Astra Polka) were tested. Both methods enabled sensitive, reproducible and specific detection of TBRV in infected plants.

Wirus czarnej pierścieniowej plamistości pomidora (Tomato black ring virus, TBRV) poraża szeroki zakres roślin gospodarczo ważnych, rośliny ozdobne oraz drzewa. Populacja wirusa jest bardzo zróżnicowana, a stężenie cząstek w roślinach zależy od takich czynników, jak: izolat, gospodarz i warunki klimatyczne. Powoduje to istotne problemy w opracowywaniu skutecznych i szybkich metod umożliwiających wykrywanie zróżnicowanych genetycznie izolatów wirusa. Stosowane dotychczas techniki do bezpośredniego wykrywania wirusa w porażonym materiale roślinnym, np. test immunoenzymatyczny ELISA, nie zawsze dawały zadowalające rezultaty. Opracowano warunki łańcuchowej reakcji polimerazy w czasie rzeczywistym (poprzedzonej odwrotną transkrypcją) oraz izotermicznej amplifikacji kwasów nukleinowych w połączeniu z serologicznym zagęszczaniem cząstek wirusa z soku roślinnego. Do badań wykorzystano 8 izolatów z robinii akacjowej (Robinia pseudoacacia L.) oraz jeden z pomidora (Solanum lycopersicum odmiana Moneymaker) i jeden z cukinii (Cucurbita pepo convar. giromontiina odmiana Astra Polka). Obie metody wykrywały wirusa w specyficzny i czuły sposób w zainfekowanych roślinach.

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