Articles
COMPARISON OF SEROLOGICAL AND MOLECULAR DETECTION METHODS FOR TESTING INDIVIDUAL AND COMPOSITE SAMPLES USING PPV-M AND PPV-T ISOLATES
Article number
1063_24
Pages
173 – 176
Language
English
Abstract
Sharka disease of stone fruit trees caused by Plum pox virus (PPV) was first described in 1968 in a limited area of Turkey, but during the last decade the disease has progressively spread to a large part of the country.
Although PPV-Rec and -D strains were found in Turkey, the most common PPV strains were detected as PPV-M and PPV-T. In this study, DAS-ELISA (5B-IVIA/AMR) monoclonal antibody) and Spot Real-time RT-PCR techniques have been evaluated in order to determine the best sampling time and ratio of PPV infected samples in non-infected-infected plant mixtures for detection of PPV-T and PPV-M strains.
Dormant buds in winter and fresh leaves in spring from PPV-infected trees were used for testing in 2012. Six repetitions were performed by single (3 leaves or buds from infected plant) or composite plant mixture samples (3 leaves or buds from infected plant + 3 leaves from healthy plant, and the other composite samples, i.e., 3+6 to 3+27). All combinations and all repetitions of composite leaf samples of both strains were detected as positive in Spot Real-time RT-PCR. However, in DAS-ELISA, the number of PPV positive samples decreased for T and M strain in 6th composite (3 infected+12 healthy leaves) and in 9th composite (3 infected+21 healthy leaves) in spring, respectively.
At least 3 repetitions in all combinations of composite samples for PPV-T and -M were found positive in dormant season by Spot Real-time RT-PCR whereas it was negative only in the last composite sample (3 infected+
27 healthy buds) of PPV-T by DAS-ELISA.
Although PPV-Rec and -D strains were found in Turkey, the most common PPV strains were detected as PPV-M and PPV-T. In this study, DAS-ELISA (5B-IVIA/AMR) monoclonal antibody) and Spot Real-time RT-PCR techniques have been evaluated in order to determine the best sampling time and ratio of PPV infected samples in non-infected-infected plant mixtures for detection of PPV-T and PPV-M strains.
Dormant buds in winter and fresh leaves in spring from PPV-infected trees were used for testing in 2012. Six repetitions were performed by single (3 leaves or buds from infected plant) or composite plant mixture samples (3 leaves or buds from infected plant + 3 leaves from healthy plant, and the other composite samples, i.e., 3+6 to 3+27). All combinations and all repetitions of composite leaf samples of both strains were detected as positive in Spot Real-time RT-PCR. However, in DAS-ELISA, the number of PPV positive samples decreased for T and M strain in 6th composite (3 infected+12 healthy leaves) and in 9th composite (3 infected+21 healthy leaves) in spring, respectively.
At least 3 repetitions in all combinations of composite samples for PPV-T and -M were found positive in dormant season by Spot Real-time RT-PCR whereas it was negative only in the last composite sample (3 infected+
27 healthy buds) of PPV-T by DAS-ELISA.
Publication
Authors
M. Gazel , C. Ulubaş Serçe, K. Çağlayan
Keywords
Sharka, apricot, plum, Turkey
Online Articles (25)