Korean J Parasitol.
2012 Jun;50(2):103-111. 10.3347/kjp.2012.50.2.103.
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
- Affiliations
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- 1Pathology Division, National Fisheries Research and Development Institute (NFRDI), Busan 619-705, Korea. ehlee1234@nfrdi.go.kr
- 2Department of Aquatic Life Medicine, Pukyong National University, Busan 608-737, Korea.
- 3Aquatic Life Disease Control Division, NFRDI, Busan 619-705, Korea.
- 4New Strategy Research Center, NFRDI, Busan 619-705, Korea.
- KMID: 1384737
- DOI: http://doi.org/10.3347/kjp.2012.50.2.103
Abstract
- 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.
Keyword
MeSH Terms
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Animals
Carps
DNA Primers/genetics
DNA, Ribosomal/chemistry/genetics
Fish Diseases/*diagnosis/parasitology
Molecular Diagnostic Techniques/*methods
Molecular Sequence Data
Myxozoa/genetics/*isolation & purification
Parasitic Diseases, Animal/*diagnosis/parasitology
RNA, Ribosomal, 18S/genetics
Real-Time Polymerase Chain Reaction/*methods
Republic of Korea
Sequence Analysis, DNA
Time Factors
Veterinary Medicine/*methods
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