Quantification of aquabirnaviruses isolated from different host species by real-time RT-PCR.
Real-time reverse transcription (RT)-PCR was developed to detect and quantify successfully aquatic birnaviruses (ABVs) from fish tissue and seawater samples. This method detected marine birnavirus (MABV) RNA in several samples, and the detection was specific for ABVs. Monitoring the MABV strain Y-6 RNA quantification by real-time RT-PCR showed replication kinetics of MABV after experimental infection in vitro. We found the quantity of ABVs isolated from different host species by using combined virus absorption in cell culture and real-time RT-PCR. Although all specimens showed no symptoms of viral infection, ABVs were detected regardless of host species. In conclusion, real-time RT-PCR was shown to be a sensitive and reliable tool to detect and quantify ABVs in cultured fish. This method is useful procedure to show details of horizontal or vertical infections by ABVs in the breeding water of aquatic animals.
Hirayama
Takeshi
T
Laboratory of Cell Structure and Function, Division of Marine Bioresource Science, Graduate School of Kuroshio Science, Kochi University, Nankoku, Kochi 783-8502, Japan.
Imajoh
Masayuki
M
Oshima
Syun-Ichirou
S
D000818:Animals; D019201:Aquabirnavirus / Q000254:growth & development / Q000302:isolation & purification*; D002460:Cell Line; D005399:Fishes / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012623:Seawater / Q000821:virology*; D012680:Sensitivity and Specificity; D014779:Virus Replication
D012367:RNA, Viral