RT-PCR and sequence analysis of Macrobrachium rosenbergii nodavirus: Indian isolate

Shekhar, M. S.; Azad, I.S.; Jithendran, K. P.

doi:10.1016/j.aquaculture.2005.07.026

Cited by 20 publications

(12 citation statements)

References 8 publications

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“…A large number of samples and reagents are required and large-scale apparatus are normally needed, thus limiting their practical applications. Several diagnostic technologies such as immunohistochemistry staining (Breton et al 1997;Zilberg and Munday 2000), cell line isolation (Chi et al 1999), In situ hybridization (Comps et al 1996) and a molecular diagnostic method (Shekhar et al 2006) have been developed for rapid diagnosis of aquaculture diseases. Among them, molecular diagnosis utilizing RT-PCR technique has been demonstrated with high sensitivity and specificity when incorporated with specific primer sets (Dhar et al 2002).…”

Section: Introductionmentioning

confidence: 99%

A microfluidic-based system using reverse transcription polymerase chain reactions for rapid detection of aquaculture diseases

Lien

Lee

Tsai

et al. 2009

Microfluid Nanofluid

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This study presents an integrated microfluidic system capable of automatically performing four reversetranscription polymerase chain reaction (RT-PCR) processes simultaneously for fast diagnosis of aquacultural diseases. This system integrates micro temperature control modules and a microfluidic control module. The micro temperature control modules have micro temperature sensors and array-type micro heaters that maintain precise and uniform temperature conditions for the RT-PCR processes. The microfluidic control module can automatically transport samples and reagents by using pneumatic micropumps, microvalves and microchannels. Moreover, by using random primers in the reverse-transcription (RT) process, the chip design is simplified and the consumption of RT products in the subsequent multiple polymerase chain reactions (PCR) is also minimized. After sample transport between the RT chamber and the PCR chambers is finished, the PCR process is then performed to amplify

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Section: Introductionmentioning

confidence: 99%

A microfluidic-based system using reverse transcription polymerase chain reactions for rapid detection of aquaculture diseases

Lien

Lee

Tsai

et al. 2009

Microfluid Nanofluid

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“…During the past few years, virus infection has also been found to cause muscle necrosis in cultured shrimp. An example is MrNV (Macrobrachium rosenbergii nodavirus) infection in freshwater prawns (Arcier et al 1999, Qian et al 2003, Bonami et al 2005, Shekhar et al 2006, causing a disease referred to as white tail disease. Another virus that infects marine shrimp is IMNV (infectious myonecrosis virus) (Poulos et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Development of in situ hybridization and RT-PCR assay for the detection of a nodavirus (PvNV) that causes muscle necrosis in Penaeus vannamei

Tang

Pantoja²,

Redman

et al. 2007

Dis. Aquat. Org.

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A nodavirus (tentatively named PvNV, Penaeus vannamei nodavirus) that causes muscle necrosis in P. vannamei was found in Belize in . From 2004, shrimp samples collected from Belize exhibited clinical signs, white, opaque lesions in the tails and histopathology similar to those of shrimps infected by infectious myonecrosis virus (IMNV). Histological examination revealed multifocal necrosis and hemocytic fibrosis in the skeletal muscle. In addition, basophilic, cytoplasmic inclusions were found in striated muscle, lymphoid organ and connective tissues. However, IMNV was not detected in these shrimps by either RT-PCR or in situ hybridization, suggesting that these lesions may be caused by another RNA virus. Thus, a cDNA library was constructed from total RNA extracted from hemolymph collected from infected shrimp. One clone (designated PvNV-4) with a 928 bp insert was sequenced and found to be similar (69% similarity when comparing the translated amino acid sequences) to the capsid protein gene of MrNV (Macrobrachium rosenbergii nodavirus). The insert of PvNV-4 was labeled with digoxigenin-11-deoxyuridine triphosphate (dUTP) and hybridized to tissue sections of P. vannamei with muscle necrosis collected in Belize and from laboratory bioassays. The samples were positive for PvNV infection. Positively reacting tissues included skeletal muscle, connective tissues, the lymphoid organ, and hemocytes in the heart and gills. In addition, we experimentally infected both P. vannamei and P. monodon with PvNV prepared from Belize samples. A nested RT-PCR assay developed from the PvNV-4 cloned sequence showed that both species are susceptible to PvNV infection. 75: 183-190, 2007 MrNV. To provide diagnostic tools, we have developed an in situ hybridization method and a nested RT-PCR assay which are specific for P. vannamei nodavirus (PvNV). MATERIALS AND METHODSShrimp, PvNV isolate, generation of inoculum and infected tissues. Shrimp taxonomy was according to Holthius (1980). The PvNV-infected tissues used in this study was obtained in 2005 in Belize from farmraised Penaeus vannamei that were exhibiting signs of muscle necrosis. The inoculum was made from the heads of these shrimp, which were stored frozen at -70°C at the University of Arizona. Soft tissues were minced, homogenized in TN buffer (20 mM Tris-HCl, 400 mM NaCl, pH 7.4), and clarified by centrifugation at 3000 × g for 30 min. The tissue homogenate was aliquoted and stored at -70°C. Prior to injection, the tissue homogenate was diluted (v/v 1:20) with 2% saline, filtered through a 0.45 µm membrane, and injected into the muscle of specific-pathogen-free (SPF) P. vannamei Kona stock, Oceanic Institute, Hawaii, Wyban et al. 1992). To generate the infected tissues for feeding experiments, the inoculated shrimp were maintained in seawater at 26 to 28°C for 4 wk. At termination, shrimp were collected and stored at -70°C. Samples of the cephalothoraces were fixed in Davidson's alcohol formalin acetic acid (AFA) fixative for 24 h, then changed to 70% ethanol ...

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“…The disease was first reported in Guadeloupe in 1997 (Arcier et al, 1999), then in Martinique (French West Indies) and in Dominican Republic (Sri Widada and Bonami, unpublished data), in the People's Republic of China and more recently, in India (Sahul Hameed et al, 2004a,b;Vijayan et al, 2005;Shekhar et al, 2006). A morphologically similar agent (MrNV-type called MMV for Macrobrachium muscle virus) was observed in Taiwan (Tung et al, 1999).…”

Section: Geographic Rangementioning

confidence: 91%

Viral diseases of the giant fresh water prawn Macrobrachium rosenbergii: A review

Bonami

Widada

2011

Journal of Invertebrate Pathology

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RT-PCR and sequence analysis of Macrobrachium rosenbergii nodavirus: Indian isolate

Cited by 20 publications

References 8 publications

A microfluidic-based system using reverse transcription polymerase chain reactions for rapid detection of aquaculture diseases

A microfluidic-based system using reverse transcription polymerase chain reactions for rapid detection of aquaculture diseases

Development of in situ hybridization and RT-PCR assay for the detection of a nodavirus (PvNV) that causes muscle necrosis in Penaeus vannamei

Viral diseases of the giant fresh water prawn Macrobrachium rosenbergii: A review

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