Early detection and estimation of infection burden by real-time PCR in rats experimentally infected with Clonorchis sinensis

• Published in: Parasitology research (1987) Vol. 109; no. 2; pp. 297 - 303
• Main Authors: Rahman, S. M. Mazidur, Bae, Young Mee, Hong, Sung-Tae, Choi, Min-Ho
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.08.2011; Springer
• Subjects: Animals; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Clonorchiasis - diagnosis; Clonorchiasis - parasitology; Clonorchis sinensis; Clonorchis sinensis - enzymology; Clonorchis sinensis - genetics; Clonorchis sinensis - isolation & purification; Cytochrome c; Cytochrome oxidase; Disease Models, Animal; DNA; Electron Transport Complex IV - genetics; Feces - parasitology; Fundamental and applied biological sciences. Psychology; General aspects; General aspects and techniques. Study of several systematic groups. Models; Genes; Health aspects; Immunology; Invertebrates; Male; Medical Microbiology; Microbiology; Original Paper; Parasitology - methods; Polymerase Chain Reaction - methods; Rats; Rats, Sprague-Dawley; Rodent Diseases - diagnosis; Rodent Diseases - parasitology; Sensitivity and Specificity; Time Factors; Worm Recovery; Internal Transcribe Spacer; Worm Burden; Fecal Examination; Single Worm; Rat; Rodentia; Plathelmintha; Parasite; Trematoda; Infection; Polymerase chain reaction; Vertebrata; Mammalia; Helmintha; Clonorchis sinensis; Invertebrata; Detection
• ISSN: 0932-0113; 1432-1955
• DOI: 10.1007/s00436-011-2253-3

A real-time PCR assay targeting the cytochrome c oxidase gene of Clonorchis sinensis was developed and its diagnostic value was investigated in experimentally infected rats. The PCR assay amplified as little as 0.1 pg of C. sinensis genomic DNA and a single egg from experimentally spiked negative feces. The assay could detect C. sinensis DNA in feces 2 weeks post-infection (PI), whereas the Kato–Katz method and formalin–ether concentration techniques could detect the eggs in the feces from 3 weeks PI. Inverse correlations between eggs per gram of feces (EPG) and Ct values were observed 4, 6, and 8 weeks PI. Worm burden was positively correlated with EPG but negatively correlated with Ct values in different groups at 8 weeks PI. Fecal real-time PCR provides a potential tool for the diagnosis of clonorchiosis before eggs can be detected by conventional methods, and may be used to estimate the worm burden in the host.