Detection of fish nocardiosis by loop-mediated isothermal amplification.
OBJECTIVE
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 detecting Nocardia seriolae which is a causative agent of fish nocardiosis, was designed.


RESULTS
A set of four primers, two inner and two outer, were designed based on the sequence of the 16S-23S ribosomal RNA internal transcribed spacer region of N. seriolae. Time and temperature conditions for detection of N. seriolae were optimized for 60 min at 65 degrees C. Other fish pathogen was not amplified by this LAMP system. The detection of N. seriola using LAMP was found to be more sensitive than that by polymerase chain reaction.


CONCLUSIONS
LAMP is a highly sensitive and rapid diagnostic procedure for detection of N. seriolae.


CONCLUSIONS
LAMP is a useful diagnostic method for fish nocardiosis.
Itano
T
T
Ehime Prefectural Fish Disease Control Center, Sakashizu, Uwajima, Ehime, Japan.
Kawakami
H
H
Kono
T
T
Sakai
M
M
D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000032:analysis*; D005393:Fish Diseases / Q000175:diagnosis*; D005399:Fishes / Q000469:parasitology; D005516:Food Microbiology; D005818:Genetic Engineering; D008969:Molecular Sequence Data; D009615:Nocardia / Q000235:genetics*; D009617:Nocardia Infections / Q000175:diagnosis*; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA; D012336:RNA, Ribosomal, 16S; D013997:Time Factors
D016054:DNA, Protozoan; C034477:RNA primers; D012336:RNA, Ribosomal, 16S; D012313:RNA