Different reactions obtained using the same DNA detection reagents for Thai and Korean hepatopancreatic parvovirus of penaeid shrimp

Abstract

Hepatopancreatic parvovirus (HPV) can cause stunted growth and death in penaeid shrimp including Penaeus monodon. We used PCR primers and a commercial DNA probe designed from HPV of Penaeus chinensis (HPVchin) to examine HPV-infected Thai P. monodon (HPVmon). We found that the PCR primers produced a 732 bp DNA amplicon rather than the 350 bp amplicon obtained with HPVchin template and that the DNA probe gave weak to variable in situ DNA hybridization results. In addition, hybridization to PCR products from HPVmon was weak compared with hybridization with PCR products from HPVchin. By contrast, the 732 bp amplicon hybridized strongly with HPVmon-infected cells by in situ hybridization but not with uninfected shrimp tissue or other shrimp viruses, thus confirming its origin from HPVmon. Cloning, sequencing and analysis of the 732 bp amplicon showed that 696 bp (excluding the primer sequences) contained 47% GC content and had only 78% homology to 701 aligned bases from a 3350 bp DNA fragment of HPVchin from GenBank. These results explain why the reagents based on HPVchin gave a different PCR product and weak hybridization results with HPVmon, and they show that multiple primers or degenerate primers may be necessary for general detection of HPV varieties. Together with previously published information on the estimated total genome sizes for HPVchin (approximately 4 kb) and HPVmon (approximately 6 kb), these data support the contention that HPVchin and HPVmon are different varieties or species, in spite of their similar histopathology.

Detection of hepatopancreatic parvovirus (HPV) in wild shrimp from India by nested polymerase chain reaction (PCR)

“…3). This was perhaps not surprising since it was previously reported that HPVmon shared 77% sequence identity to HPV from P. chinensis (HPVchin) (Phromjai et al 2001, Roekring et al 2002. These constitute sequence variations in HPV detected in different hosts from different geographical locations and it raises the question as to whether these differences are related to host species or regional origin.…”

Complete nucleotide sequence analysis of a Korean strain of hepatopancreatic parvovirus (HPV) from Fenneropenaeus chinensis

“…Heavy infection may result in stunted growth, reduced production, and death [3,[14][15][16]. HPV particles, which can be used as sources for virus particles or virus-associated DNA isolation, are associated with hepatopancreatic tissue, pereiopods, and fecal matter of infected shrimp [11,[14][15][16][17][18]. In contrast, another small shrimp-infecting parvovirus, infectious hypodermal, and hepatopoietic necrosis virus (IHHNV), is similar to HPV in size, shape, and genome organization, but infects multiple organs of ectodermal and mesodermal origin and not the midgut [19].…”

Generation of monoclonal antibodies specific to hepatopancreatic parvovirus (HPV) from Penaeus monodon

“…Detection was traditionally dependent upon the histological demonstration of basophilic intranuclear inclusions in tubular epithelial cells of the hepatopancreas (HP) by hematoxylin and eosin (H&E) or Giemsa staining (Lightner 1996). DNA probes and PCR primers have been developed and proven to be powerful tools for diagnosis of shrimp HPV infection (Sukhumsirichart et al 1999, Pantoja & Lightner 2000, Phromjai et al 2001. Under optimized conditions, DNA approximately equivalent to that in 300 viral particles could be detected (Pantoja & Lighner 2000).…”

“…For example, HPV in P. chinensis from Korea (Bonami et al 1995), P. monodon from Thailand (Sukhumsirichart et al 1999) and Macrobrachium rosenbergii from Malaysia (Lightner et al 1994) are different. PCR primers designed for HPV from P. chinensis were not particularly efficient for PCR amplification of HPV DNA from P. monodon (Phromjai et al 2001) and probes designed from the same source did not hybridize with DNA of HPV from M. rosenbergii (Lightner et al 1994).…”