Genome-based detection methods of Macrobrachium rosenbergii nodavirus, a pathogen of the giant freshwater prawn, Macrobrachium rosenbergii dot-blot, in situ hybridization and RT-PCR.
The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral diseases as in this case early diagnosis is a critical factor in containing disease outbreaks. Three complementary genome-based methods were developed for the detection of Macrobrachium rosenbergii nodavirus (MrNV), i.e. dot-blot hybridization, in situ hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR). Detection limits were established for dot-blot hybridization and RT-PCR and are c. 7 fg and 8 pg of viral RNA, respectively. In situ hybridization indicated that infection was confined to the striated muscle tissue. As a result of its sensitivity, RT-PCR can be used for in-depth investigations to examine the extent of the viral infection and establish the onset of infection in hatcheries. The application of RT-PCR on samples collected from prawn farms in China showed the possible use of this method in routine health monitoring.
Sri Widada
J
J
Pathogens and Immunity, CNRS, Université Montpellier II, Place Eugène Bataillon, Montpellier, France. widada@crit.univ-montp2.fr
Durand
S
S
Cambournac
I
I
Qian
D
D
Shi
Z
Z
Dejonghe
E
E
Richard
V
V
Bonami
J R
JR
D000818:Animals; D017756:Aquaculture / Q000379:methods*; D002681:China; D017931:DNA Primers; D015151:Immunoblotting; D017403:In Situ Hybridization; D018482:Muscle, Skeletal / Q000821:virology; D030381:Nodaviridae / Q000235:genetics*; D033541:Palaemonidae / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers