Detection of white spot syndrome virus in shrimp by loop-mediated isothermal amplification.
Loop-mediated isothermal amplification (LAMP) method is a novel, sensitive and rapid technique which can be applied for disease diagnosis in aquaculture. Using the LAMP method, a highly specific and sensitive diagnostic system for white spot syndrome virus (WSSV) detection was designed. A set of four primers, two outer and two inner primers, were designed from WSSV genome DNA. Reaction time and temperatures were optimized for 60 min at 65 degrees C, respectively. The detection limit (DL) using the LAMP method was up to 1 fg, when compared to 10 fg by nested polymerase chain reaction (PCR). Thus, standardized LAMP procedure was used to detect the presence of WSSV in the heart, stomach and lymphoid organ from infected shrimp. The study has developed a diagnostic procedure which is a rapid and highly sensitive for WSSV detection in shrimp.
Kono
Tomoya
T
Faculty of Agriculture, Miyazaki University, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
Savan
Ram
R
Sakai
Masahiro
M
Itami
Toshiaki
T
D000818:Animals; D017756:Aquaculture; D017931:DNA Primers; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D033561:Penaeidae / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013696:Temperature
D017931:DNA Primers; D004279:DNA, Viral