Immunocapture PCR Detection of ToLCNDV from Plant Extract by using Heterologous Virus Coat Protein Antisera
Immunocapture PCR Detection of ToLCNDV from Plant Extract by using Heterologous Virus Coat Protein Antisera
Zafar Iqbal1* and Muhammad Khurshid2
ABSTRACT
Immunocapture-PCR (IC-PCR) is very versatile technique that has the potential to expedite the detection of plant viruses from plant species containing various forms of PCR amplification inhibitors. In current study, IC-PCR was used for the detection of Tomato leaf curl New Delhi virus (ToLCNDV) from plant extracts of Nicotiana benthamiana plants inoculated with ToLCNDV. To immunocapture the ToLCNDV, two antisera raised against Tomato yellow leaf curl virus-coat protein (TYLCV-CP) and African cassava mosaic virus-coat protein (ACMV-CP) were used. However, immunocapture of ToLCNDV could only be achieved by using the TYLCV-CP antisera followed by PCR detection by using specific and degenerate primers. ToLCNDV is geographically wide spread Begomovirus (Family Geminiviridae), considered as a close relative of TYLCV, and cause severe losses for many economical important crops in tropical and subtropical regions of the world. This is the first report of the use of heterologous polyclonal antibodies (PAbs), raised against a geographically distinct begomovirus, ToLCNDV. The described results have shown to exhibit a many fold increase detection sensitivity and reproducibility hence, can be adopted for routine detection of ToLCNDV from plant extracts.
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June
2017
Vol. 49, Iss. 3, Pages 761-1149
