A yellow head virus gene probe: nucleotide sequence and application for in situ hybridization

Diseases of Aquatic Organisms

Donald V. Lightner¹  Kathy F.-J. Tang¹ 

关键词: Penaeid shrimp;    Yellow head virus;    In situ hybridization;    RT-PCR;   

DOI  :  10.3354/dao035165

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: A portion of the genome of yellow head virus (YHV) of penaeid shrimp was cloned and the cDNA fragment (1161 bp) was designated clone 3-27. The fragment was labeled with digoxigenin and hybridized in situ to tissue sections of YHV-infectedPenaeus vannamei. Positively reacting tissues included those of the lymphoid organ, cuticular epithelium, and gills. In addition, connective tissue of hepatopancreas, heart, antennal gland, hematopoietic organ, nerve tract, midgut cecum and musclereacted to the probe. The probe was highly specific since it hybridized only to tissues from YHV-infected shrimp. It did not react to those of uninfected shrimp or shrimp infected with WSSV (white spot syndrome virus), IHHNV (infectious hypodermal andhematopoietic necrosis virus), or TSV (Taura syndrome virus). The clone was sequenced, and primers were synthesized for rapid detection of YHV in hemolymph using RT-PCR (reverse transcription-polymerase chain reaction). The strand that constituted theviral sequence in the cDNA was also determined via RT-PCR and in situ hybridization with a single-stranded RNA (ssRNA) probe.

【 授权许可】

Unknown