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Title: Identification of sacbrood virus disease in honeybee, Apis mellifera L. by using ELISA and RT-PCR techniques
Authors: Rana, Rakesh
Rana, B S
Kaushal, Nimisha
Kumar, Deep
Kaundal, Priyanka
Rana, Kiran
Khan, M Azhar
Gwande, S J
Sharma, H K
Keywords: Amplicons;Antiserum;Apis mellifera L;ELISA;Nucleotides;Primers;Prepupae;RT-PCR;SBV
Issue Date: Jul-2011
Publisher: NISCAIR-CSIR, India
Abstract: Studies were conducted on the identification of sacbrood virus (SBV) through ELISA and RT-PCR techniques in diseased prepupae of Apis mellifera L. collected from Himachal Pradesh, India. The ELISA kit was prepared from antiserum specific to Thai SBV (TSBV). SBV in the crude samples was detected at 1:1000 titre of the kit. Five pairs of oligonucleotide primers were used to target SBV genome through RT-PCR technique. Presence of SBV was confirmed with 3 primers, viz., Sb1 f/r, Sb3 f/r and Sb5 f/r. Amplification of the genome produced distinct bands/amplicons of expected molecular sizes, i.e., approximately 487, 267 and 597 bp. The amplicon Sb3 was sequenced which yielded a fragment of 822 nucleotides length. The nucleotide sequence was submitted to GenBank database, NCBI, USA under accession number EU232663. Phylogenetically, present SBV was found closer to Nepal-1, Nepal-4, followed by India-2, Nepal-2 and Germany-7 strains.
Page(s): 274-284
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.10(3) [July 2011]

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