Molecular identification and real-time quantitative PCR (qPCR) for rapid detection of Thelohanellus kitauei, a Myxozoan parasite causing intestinal giant cystic disease in the Israel carp.
Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.
Pathology Division, National Fisheries Research and Development Institute, Busan 619-705, Korea.
Jung Soo
JS
Seo
Eun Ji
EJ
Jeon
Moo Sang
MS
Kim
Sung Ho
SH
Woo
Jin Do
JD
Kim
Sung Hee
SH
Jung
Myoung Ae
MA
Park
Bo Young
BY
Jee
Jin Woo
JW
Kim
Yi-Cheong
YC
Kim
Eun Hye
EH
Lee
D000818:Animals; D002347:Carps; D017931:DNA Primers / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005393:Fish Diseases / Q000175:diagnosis* / Q000469:parasitology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D055764:Myxozoa / Q000235:genetics / Q000302:isolation & purification*; D010273:Parasitic Diseases, Animal / Q000175:diagnosis* / Q000469:parasitology; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D056910:Republic of Korea; D017422:Sequence Analysis, DNA; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods*
D017931:DNA Primers; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S