Rapid diagnosis of Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD) in salmonid fish by a novel DNA amplification method, loop-mediated isothermal amplification (LAMP).
A new molecular diagnostic assay was developed for detection of Tetracapsuloides bryosalmonae the causative agent of proliferative kidney disease (PKD) in salmonid fish using a loop-mediated isothermal amplification method (LAMP). The PKD-LAMP assay amplifies the T. bryosalmonae DNA extracted from infected kidney, under constant temperature of 65 degrees C within 1 h. The required equipment for DNA amplification is only a water bath. The amplification products were detected visually by using SYBR green I dye, which turns green in the presence of amplified products and remains orange in its absence, and by electrophoresis without any difference in the sensitivity of both methods. The developed PKD-LAMP assay demonstrated an exceptionally higher sensitivity than the conventional PCR. PKD-LAMP assay was found to be 100-fold more sensitive than the PCR assay. The developed assay is simple, rapid, cost-effective, specific and highly sensitive. The assay is also characterized by its field applicability, as it does not require the use of sophisticated equipment or skilled personnel.
El-Matbouli
Mansour
M
Institute of zoology, fish biology and fish diseases, Faculty of veterinary medicine, University of Munich, Kaulbachstr. 37, 80539 Munich, Germany. El-Matbouli@lmu.de
Soliman
Hatem
H
D000818:Animals; D052160:Benzothiazoles; D016054:DNA, Protozoan / Q000032:analysis*; D003959:Diamines; D056890:Eukaryota / Q000235:genetics / Q000302:isolation & purification*; D005393:Fish Diseases / Q000175:diagnosis* / Q000469:parasitology; D007668:Kidney / Q000469:parasitology; D007674:Kidney Diseases / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D008099:Liver / Q000469:parasitology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals; D011529:Protozoan Infections, Animal / Q000175:diagnosis*; D011804:Quinolines; D012487:Salmonidae; D012680:Sensitivity and Specificity; D013154:Spleen / Q000469:parasitology
D052160:Benzothiazoles; D016054:DNA, Protozoan; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I