Citrus leprosis virus C (CiLV-C) causes damage in citrus production in the South and CentralAmerica. Since
closely related types of citrus viruses have recently been described monoclonal antibodies (MAbs) are
needed for accurate and sensitive diagnosis of CiLV-C. In this study, MAbs to the expressed coat protein
of CiLV-C were produced for serological detection of CiLV-C in crude extracts of infected tissues in double
antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA), dot blot immunosorbent assays
(DBIA) and immuonocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) procedures.
Monoclonal antibodies were developed in mice to the purified expressed coat protein of CiLV-C. The
published standard protocols of DAS-ELISA, DBIA and IC-RT-PCR were followed for the detection of coat
protein p29 of CiLV-C in the crude extracts of CiLV-C infected tissues. Two monoclonal antibodies, designated
G10 and C11, were identified from four potential candidates for the specific and sensitive detection
of coat protein p29 of CiLV-C in the crude citrus extracts of CiLV-C infected tissues in DAS-ELISA, whereas
G10 was also selected based on performance for use in the DBIA and IC-RT-PCR diagnostic assays. Sensitivity
analysis comparing the three methods for detection of coat protein p29 of CiLV-C determined that
IC-RT-PCR was more sensitive than DAS-ELISA and DBIA. The creation of MAbs to CiLV-C allows for the
sensitive and accurate detection of the virus from CiLV-C infected citrus leaf tissues. Successful detection
of the virus in three diagnostic assays formats provides flexibility to diagnosticians who can use either
ELISA or DBIA for screening large numbers of samples, and IC-RT-PCR for rapid, sensitive confirmation
testing
Citrus leprosis virus C (CiLV-C) causes damage in citrus production in the South and CentralAmerica. Sinceclosely related types of citrus viruses have recently been described monoclonal antibodies (MAbs) areneeded for accurate and sensitive diagnosis of CiLV-C. In this study, MAbs to the expressed coat proteinof CiLV-C were produced for serological detection of CiLV-C in crude extracts of infected tissues in doubleantibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA), dot blot immunosorbent assays(DBIA) and immuonocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) procedures.Monoclonal antibodies were developed in mice to the purified expressed coat protein of CiLV-C. Thepublished standard protocols of DAS-ELISA, DBIA and IC-RT-PCR were followed for the detection of coatprotein p29 of CiLV-C in the crude extracts of CiLV-C infected tissues. Two monoclonal antibodies, designatedG10 and C11, were identified from four potential candidates for the specific and sensitive detectionof coat protein p29 of CiLV-C in the crude citrus extracts of CiLV-C infected tissues in DAS-ELISA, whereasG10 was also selected based on performance for use in the DBIA and IC-RT-PCR diagnostic assays. Sensitivityanalysis comparing the three methods for detection of coat protein p29 of CiLV-C determined thatIC-RT-PCR was more sensitive than DAS-ELISA and DBIA. The creation of MAbs to CiLV-C allows for thesensitive and accurate detection of the virus from CiLV-C infected citrus leaf tissues. Successful detectionof the virus in three diagnostic assays formats provides flexibility to diagnosticians who can use eitherELISA or DBIA for screening large numbers of samples, and IC-RT-PCR for rapid, sensitive confirmationtesting
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