On-site detection of Citrus tristeza virus (CTV) by lateral flow immunoassay using polyclonal antisera derived from virions produced by a recombinant CTV
| Autor: | Subhas Hajeri, Vijayanandraj Selvaraj, Chandrika Ramadugu, Yogita Maheshwari, Raymond Yokomi, Manjunath L. Keremane |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Antiserum biology medicine.diagnostic_test Nicotiana benthamiana Citrus tristeza virus Plant Science biology.organism_classification 01 natural sciences Virology Molecular biology law.invention 03 medical and health sciences 030104 developmental biology law Polyclonal antibodies Insect Science Complementary DNA Immunoassay Plant virus Recombinant DNA medicine biology.protein 010606 plant biology & botany |
| Zdroj: | Phytoparasitica. 45:333-340 |
| ISSN: | 1876-7184 0334-2123 |
| DOI: | 10.1007/s12600-017-0591-0 |
| Popis: | Maintenance of virus-free citrus in nurseries and orchards is essential to control the spread of aphid-transmitted Citrus tristeza virus (CTV). To this end, a lateral flow immunoassay (LFIA) was developed to detect CTV that was user-friendly and field-deployable. Polyclonal antisera was generated in rabbit against purified virions from Nicotiana benthamiana inoculated with an infectious recombinant cDNA clone of CTV (rCTV). Affinity purified rCTV IgG (1 mg/ml) and Mouse IgG (0.5 mg/ml) were conjugated with gold nanoparticles and coated onto a glass fiber membrane conjugate pad. Affinity purified rCTV IgG and anti-mouse IgG were used as test line and control line, respectively on the test strip. The LFIA detected CTV within 10 min and was sensitive up to a 1:80 dilution of crude plant sap extract. The LFIA was validated using CTV strains T30, T36, VT and RB from greenhouse and field sources and results were 100% in agreement with Enzyme-Linked Immunosorbent Assay and Reverse Transcription quantitative PCR. The immunostrip readily detected CTV after six months in storage in a sealed bag with silica gel at room temperature indicating good stability. The LFIA test strips offered a cost-effective tool to detect CTV in the field or nursery by non-skilled personnel without laboratory equipment. |
| Databáze: | OpenAIRE |
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