Detection of koi herpesvirus in common carp, Cyprinus carpio L., by loop-mediated isothermal amplification.
Loop-mediated isothermal amplification (LAMP) is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. In this study, using the LAMP method, a protocol for koi herpes virus (KHV) detection in common carp was designed. A set of four primers, two inner and two outer, were designed based on the sequence of the thymidine kinase (tk) gene of KHV. Time and temperature conditions for detection of KHV were optimized for 60 min at 65 degrees C. The detection limit using LAMP was found to be similar to that by polymerase chain reaction. In this study, we have developed a highly sensitive and rapid diagnostic procedure for detection of KHV infection in common carp.
United Graduate School of Agriculture Sciences, Kagoshima University, Kagoshima, Japan.
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I
Gunimaladevi
T
T
Kono
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MN
Venugopal
M
M
Sakai
D000818:Animals; D001483:Base Sequence; D002347:Carps; D017931:DNA Primers; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology*; D006564:Herpesviridae / Q000235:genetics*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary; D013696:Temperature; D013937:Thymidine Kinase / Q000235:genetics; D013997:Time Factors
D017931:DNA Primers; D013937:Thymidine Kinase