PRINT CAPTURE PCR – A SIMPLE TECHNIQUE FOR THE DETECTION OF TOMATO LEAF CURL NEW DELHI VIRUS – CAUSAL AGENT OF POTATO APICAL LEAF CURL DISEASE IN INDIA

Abstract

A simple, quick, economical, sensitive and reliable polymerase chain reaction-based method for the detection of Tomato leaf curl New Delhi virus (ToLCNDV) causing potato apical leaf curl disease in potato was developed. The method is called print-capture PCR (PC-PCR). It does not involve costly and time consuming viral DNA extraction used in the conventional-PCR detection. The technique involves adsorption of the virus on to nylon membrane directly from the freshly cut end of the virus infected leaf tissue (tissue printing). The viral DNA is then eluted from the print in sterile distilled water at 95°C and used to amplify the coat protein gene of the virus. The method is as sensitive as the conventional PCR and is suitable for routine detection of the virus in potato samples.