Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Reexamination Certificate
1998-04-16
2001-02-20
Myers, Carla J. (Department: 1655)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
C435S006120, C435S091200, C435S235100, C536S024320, C536S024330, C536S023720, C536S025400, C424S204100
Reexamination Certificate
active
06190862
ABSTRACT:
FIELD OF INVENTION
This invention relates to the identification, purification and detection of a new infectious viral agent in arthropods, especially shrimps. The virus is named as WSBV (Baculovirus associated with white spot syndrome).
BACKGROUND OF THE INVENTION
Recently, disease outbreaks have caused mass mortality among cultured penaeid shrimps in Asian countries. Since 1992, outbreaks of a new disease leading to serious mortality among populations of cultured kuruma shrimp (Penaeus japonicus) have occurred in northern Taiwan. The disease is characterized by obvious white spots on the carapace, appendages and the inside surface of the body, and cumulative mortality reaches 100% within 2-7 days. The diseased shrimps also display signs of lethargy and reddish coloration of the hepatopancreas. In 1993, white spot syndrome (W.S.S.) in cultured giant tiger prawn (P. monodon) and redtail prawn (P. penicillatus) was observed. Serious damage to penaeid shrimp production by W.S.S. in Taiwan has been reported (Tung et al. personal communication).
An epizootiological survey of kuruma shrimp in Japan reports similar findings (Nakano et al., Fish Pathology, 29 (2):135-139, 1994). According to the evidence from electron microscopy and the results of challenge tests with the filtrate from diseased shrimp lymphoid organs, the causative agent was a virus that was temporarily designated RV-PJ, a rod-shaped nuclear virus of Penaeus japonicus (Inouye et al., Fish Pathology, 29 (2):149-158, 1994; Takahashi et al., Fish Pathology, 29 (2):121-125, 1994).
To date, the prevalence of baculoviruses in cultured penaeid shrimps has been well documented (Lightner et al., Aquaculture, 32: 209-233, 1983). Among these penaeid baculoviruses, monodon baculovirus (MBV), baculoviral mid-gut necrosis virus (BMNV) and Baculovirus penaei (BP) were considered to be the most important because they have on occasions caused serious losses in infected shrimp populations (Couch, Nature 247 (5438): 22-231, 1974; J. Invertebr. Pathol., 24:311-331, 1974; Lightner & Redman, J. Invertebr. Pathology, 38: 299-302, 1981; Lightner et al. (1983), supra; Lightner et al., 1987; Sano et al., Fish Pathology, 15: 185-191, 1981).
Baculovirus-like viral particles were observed in the spontaneously diseased penaeid shrimp with W.S.S. (Tung et al., personal communication). This virus may possibly be the main causative agent for the W.S.S. that has occurred in Taiwan in penaeid shrimps. To prevent the spread of W.S.S. in shrimps, thereby rescuing the financial losses caused by this viral disease, it is necessary to identify the actual causative agent and then to develop a diagnostic method that is easy, accurate and not time-consuming in the detection of W.S.S. without sacrificing the whole subjects tested.
SUMMARY OF THE INVENTION
This invention is based on the finding of a new causative agent responsible for the incidence of white spot syndrome in penaeid shrimps. The causative agent has been isolated and purified and found to be a non-occluded rod-shaped virus particle, which is enveloped, 330±20 nm in length and 87±7 nm in diameter. This virus is determined to be a member of genus NOB (Non-Occluded Baculovirus) of the subfamily Nudibaculovirinae of Baculoviridae and the present isolate is designated as PMNOBIII, and as WSBV (Baculovirus associated with White Spot syndrome) to indicate PmNOBIII related agents. A WSBV genomic DNA library was constructed and based upon the sequence of one of the cloned WSBV DNA fragments, a WSBV specific primer set for PCR to detect the WSBV infection in penaeid shrimps has been developed. By PCR with the WSBV specific primer set, it was demonstrated that the causative agents of white spot syndrome in different shrimp species are in fact closely related. The results of the present invention provide an effective diagnostic tool for screening WSBV infection in animal host organisms, in particular shrimps, which tool may be extremely important in preventing the further spread of this viral disease. An easy, sensitive and specific ready-to-use diagnostic product, which includes primers established based on the nucleotide sequence of a unique genomic DNA clone derived from WSBV, can be developed for the detection of the presence of WSBV and to halt the further spread of this viral disease. The WSBV (also called PmNOBIII) described herein was deposited under the terms of the Budapest Treaty on Jan. 11, 1996, with the China Center for Type Culture Collection, Wuhan University, Luo Jia Shan, Wuhan, Hubei, 430072, People's Republic of China, where the deposit was given accession number CCTCC-V96001.
Features and advantages of the present invention will become apparent in the following detailed description of the preferred embodiments, with reference to the accompanying drawings, of which:
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Kou Guang-Hsiung
Lo Chu-Fang
Wang Chung-Hsiung
Fish & Richardson P.C.
Myers Carla J.
National Science Council
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