A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss).
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol was developed for detection of infectious hematopoietic necrosis virus (IHNV) RNA in rainbow trout (Oncorhynchus mykiss). A set of four primers, two outer and two inner primers for the RT-LAMP and the LAMP assay, were designed based on the sequence of G-protein of IHNV. Time and temperature conditions were optimized for 60 min at 63 degrees C for both RT-LAMP and LAMP protocols. The detection limit was found to be similar for both RT-LAMP and LAMP. When the sensitivity of RT-LAMP and LAMP were compared with conventional nested PCR, a10-fold higher sensitivity was seen for the LAMP protocols.
Gunimaladevi
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United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan.
Kono
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Lapatra
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Sakai
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D000818:Animals; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D005393:Fish Diseases / Q000821:virology*; D029142:Infectious hematopoietic necrosis virus / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D017686:Oncorhynchus mykiss / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D018353:Rhabdoviridae Infections / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral