אסיף מאגר המחקר החקלאי

Evaluating the use of immunocapture and sap-dilution PCR for the detection of prunus necrotic ringspot virus

Year:

1998

Source of publication :

Authors :

Volume :

472

Co-Authors:

Krisbai, L., Plant Health and Soil Conservation Station, Budapest, Hungary
Kölber, M., Plant Health and Soil Conservation Station, Budapest, Hungary

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Abstract:

Reaction conditions were investigated for the detection of PNRSV in peach trees using immunocapture-polymerase chain reaction (IC-PCR) with avian myeloblastosis virus reverse-transcriptase (AMV-RT). Incubation of the RT reaction at 46°C resulted in higher levels of amplified products as compared with the recommended 37°C. Preheating the reaction mixture at 55°C for 5 minutes further improved PCR yields. As an alternative to IC, PNRSV could be detected by RT-PCR directly in plant sap. Undiluted sap was negative in PCR, presumably due to the presence of PCR inhibitors. Dilution of sap (at least 1:50) resulted in a successful PCR amplification of the virus-specific DNA fragment. This direct procedure can be easily applied for large-scale testing, however, IC-PCR, which also avoids the inhibitors effect, proved to be more sensitive and suitable for low virus titers.

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