20933014
not annotated - annotated - LINNAEUS only
Monoclonal antibody-based triple antibody sandwich-enzyme-linked immunosorbent assay and immunocapture reverse transcription-polymerase chain reaction for Odontoglossum ringspot virus detection.
Odontoglossum ringspot virus (ORSV) infects numerous commercially important orchids and causes significant losses worldwide. The coat protein (CP) gene of ORSV was cloned and expressed in Escherichia coli by using the pET-32a expression vector, and the expression of recombinant protein was confirmed by Western blotting using anti-ORSV antibodies. The recombinant protein was purified using Ni-NTA agarose, and the purified protein was used as an immunogen to produce monoclonal antibodies (MAbs) and polyclonal antibodies (PAbs). Five murine MAbs against ORSV CP were obtained. Among them, two MAbs (6B4 and 1D1) also reacted with TMV CP. The triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) and immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) methods using the MAb (8A5) were then developed for sensitive, specific, and rapid detection of ORSV. TAS-ELISA and IC-RT-PCR could detect ORSV in the infected leaf saps with dilutions of 1:10,240 and 1:81,920 (w/v, g mL(-1)), respectively. TAS-ELISA and IC-RT-PCR detections indicated that ORSV was prevalent in orchids in the Zhejiang Province of China.
Ann file
T1 Species 155 183 Odontoglossum ringspot virus
N1 Reference T1 Taxonomy:12238
T2 Species 197 225 Odontoglossum ringspot virus
N2 Reference T2 Taxonomy:12238
T3 Species 227 231 ORSV
N3 Reference T3 Taxonomy:12238
T4 Species 352 356 ORSV
N4 Reference T4 Taxonomy:12238
T5 Species 385 401 Escherichia coli
N5 Reference T5 Taxonomy:562
T6 Out-of-scope 529 533 ORSV
N6 Reference T6 Taxonomy:12238
T7 Species 754 758 ORSV
N7 Reference T7 Taxonomy:12238
T8 Species 1087 1091 ORSV
N8 Reference T8 Taxonomy:12238
T9 Species 1130 1134 ORSV
N9 Reference T9 Taxonomy:12238
T10 Species 1282 1286 ORSV
N10 Reference T10 Taxonomy:12238