Q: can you tell me when was this kit (antigen/antibody) developed for the first time by using which strains?
A: The polyclonal antibody for coating and monoclonal for detection used in the ELISA were developed at the USDA laboratory against a BlScV viral isolate from Oregon State, USDA, during the 1980s. The antibodies have been using for developing the ELISA at our labs and this ELISA kit has been using for detecting BlScV in plant tissues in many countries around the world for many years. The sensitivity is relatively high, and the virus can be detected at dilutions of 1:90-1:270 in ground infected plant tissues.
Q: Do they come in removable strips of eight wells or not? It works better for us if they are removable strips as we do not wait for batch testing
A: It contains a 96-well plate with 12 strips of 8 wells.
Q: I am interested in knowing the sensitivity capability of these tests.
A: Our ELISA for Blueberry Scorch Virus (BlScV) is not a quantality test. The sensitivity of this assay is only based on the sample dilution as shown in our webpage. At our lab, the virus can be detected at dilution of 1:30-1:270 in BIScV infected samples depending on the different plant samples.
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References
Aphid population dynamics on highbush blueberry in relation to the spread of Blueberry scorch virus in Piedmont (NW Italy)
In 2004, infections of Blueberry scorch virus (BIScV) were detected for the first time outside North America on Vaccinium corymbosum L. (Ericaceae) in Piedmont (NW Italy). In its original area, this vir-us is transmitted by Ericaphis scammelli (Mason) (Homoptera Aphididae) in a nonpersistent way. In 2005-2006, field surveys were fortnightly carried out to monitor aphid populations in a seriously infected highbush blueberry crop in Costigliole Saluzzo (province of Cuneo). Moreover, to evaluate the aphid transmission ability, during each survey from 10 to 100 specimens were transferred from surely infected plants to virus-free potted plants. In both years, E. scammelli was the predominant aphid in the investigated highbush blueberry crop. Infestation levels fluctuated from 3% to 64% of infested buds in 2005, and from 0.3% to 35% in 2006. In transmission trials, E. scammelli proved not to be a very efficient vector of BIScV because only 2/55 plants infested with aphids field-collected on infected plants were positive with the ELISA test.
Complete nucleotide sequence and molecular probing of Potato virus S genome
Complete genomes of three isolates of Potato virus S (PVS) were cloned and sequenced. The PVS ORF-1 was characterized for the first time. It encodes a putative replication protein (RPT) that shares the highest homology (about 52%) with that of Blueberry scorch virus (BIScV). ORF-1 motifs, characteristic for carlaviruses were found for methyltransferase (MTR), helicase (HEL) and RNA-dependent RNA polymerase (RdRp). The complete sequence of PVS genome enabled to develop an irnmunocapture RT-PCR probing of the PVS genome. Using this system, the sequence variability of 11 genome zones was examined for 34 PVS isolates including 15 PVS-CS variants that caused a systemic infection in Chenopodium quinoa. A broad variability between PVS isolates and diverse sequence variants was found. cDNA fragments covering the coat protein (CP) leader and CP-coding region (approx. 420 bp) were pooled for PVS-O and Chenopodium-systemic PVS isolates (PVS-CS) and corresponding cDNA libraries were screened for sequence variants. Both cDNA pools differred mainly in the 5'-end of the CP gene. Methionine at the position 17 in combination with serine at the position 34 were frequently associated with the CS character of PVS. In general, hydrophobic and polar amino acids were characteristic for the positions 17 and 34, respectively in PVS-CS isolates. Genome probing and evolutionary distances suggested that the PVS-CS isolates analyzed were close to the ordinary European isolates of ordinary strain of PVS (PVS-O) but distant to the original Andean strain of PVS (PVS-A).
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