Development of Dot-ELISA and Colloidal Gold Immunochromatographic Strip for Rapid and Super-Sensitive Detection of Plum Pox Virus in Apricot Trees

Mengmeng Guo; Duo Qi; Jinxi Dong; Saiyu Dong; Xiuling Yang; Yajuan Qian; Xueping Zhou; Jianxiang Wu

doi:10.3390/v15010169

Development of Dot-ELISA and Colloidal Gold Immunochromatographic Strip for Rapid and Super-Sensitive Detection of Plum Pox Virus in Apricot Trees

Viruses. 2023 Jan 5;15(1):169. doi: 10.3390/v15010169.

Authors

Mengmeng Guo¹, Duo Qi¹, Jinxi Dong^{1

2}, Saiyu Dong¹, Xiuling Yang³, Yajuan Qian¹, Xueping Zhou^{1

3}, Jianxiang Wu^{1

2}

Affiliations

¹ State Key Laboratory of Rice Biology, Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China.
² Hainan Institute, Zhejiang University, Sanya 572025, China.
³ State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

Abstract

Plum pox virus (PPV) is a causal agent of the stone fruit tree sharka disease that often causes enormous economic losses. Due to its worldwide distribution and economic importance, rapid and reliable diagnostic technologies are becoming increasingly important for successful management of sharka disease. In this study, we have produced two super-sensitive and specific anti-PPV monoclonal antibodies (i.e., MAbs 13H4 and 4A11). Using these two MAbs, we have now developed a dot enzyme-linked immunosorbent assay (dot-ELISA) and a colloidal gold immunochromatographic strip (CGICS) assay. These two technologies can be used to quickly and reliably detect PPV. The results of these sensitivity assays confirmed that the dot-ELISA and CGICS assays could detect PPV infection in apricot tree leaf crude extracts diluted up to 1:5120 and 1:6400 (w/v), respectively. Further analyses using field-collected apricot tree leaf samples showed that the detection endpoint of the dot-ELISA was ~26 times above that obtained through RT-PCR, and the CGICS was as sensitive as RT-PCR. This present study is to broaden the knowledge about detection limits of dot-ELISA and CGICS for PPV monitoring. We consider that these newly developed dot-ELISA and CGICS are particularly useful for large scale PPV surveys in fields.

Keywords: RT-PCR; apricot tree; colloidal gold immunochromatographic strip; dot-ELISA; monoclonal antibody; plum pox virus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Monoclonal
Enzyme-Linked Immunosorbent Assay / methods
Gold Colloid
Plant Diseases
Plum Pox Virus*
Prunus armeniaca*

Substances

Gold Colloid
Antibodies, Monoclonal

Grants and funding

This research was financially supported by the National Key R&D Program of China (grant no. 2019YFD1001800), the National Natural Science Foundation of China (grant no. 31871929 and 31972234) and the Hainan Province Science and Technology Special Fund (grant no. ZDYF2022XDNY240).