Abstract
BackgroundClonorchiasis is prevalent in the Far East, and a major health problem in endemic areas. Infected persons may experience, if not treated, serious complications such as bile stone formation, pyogenic cholangitis, and even cholangiocarcinoma. Early diagnosis and treatment are important to prevent serious complications and, therefore, the simple and reliable diagnostic method is necessary to control clonorchiasis in endemic areas, where resources for the diagnosis are limited.Methodology/Principle findingsThe loop-mediated isothermal amplification (LAMP) assay has been applied for the detection of Clonorchis sinensis DNA. Six primers targeting eight locations on the cytochrome c oxidase subunit 1 gene of C. sinensis were designed for species-specific amplification using the LAMP assay. The LAMP assay was sensitive enough to detect as little as 100 fg of C. sinensis genomic DNA and the detection limit in 100 mg of stool was as low as one egg. The assay was highly specific because no cross-reactivity was observed with the DNA of other helminths, protozoa or Escherichia coli. Then, LAMP assay was applied to human fecal samples collected from an endemic area of clonorchiasis in Korea. Using samples showing consistent results by both Kato-Katz method and real-time PCR as reference standards, the LAMP assay showed 97.1% (95% CI, 90.1–99.2) of sensitivity and 100% (95% CI, 92.9–100) of specificity. In stool samples with more than 100 eggs per gram of feces, the sensitivity achieved 100%.ConclusionsTo detect C. sinensis in human fecal samples, the LAMP assay was applied and achieved high sensitivity and specificity. The LAMP assay can be utilized in field laboratories as a powerful tool for diagnosis and epidemiological survey of clonorchiasis.
Highlights
Clonorchiasis is an important human parasitic infection and is highly prevalent in eastern Asian countries, including China, Korea, and Vietnam [1, 2]
The loop-mediated isothermal amplification (LAMP) assay can be utilized in field laboratories as a powerful tool for diagnosis and epidemiological survey of clonorchiasis
Institutional review of this study was waivered because this study used anonymized stool samples that were randomly selected from the pool of stool samples of the residents of an endemic area of clonorchiasis in Korea, which had been obtained from the previous studies [28, 29]
Summary
Clonorchiasis is an important human parasitic infection and is highly prevalent in eastern Asian countries, including China, Korea, and Vietnam [1, 2]. In Korea, the Clonorchis sinensis egg positive rate in the general population is 1.9%, and approximately 1 million people are estimated to be infected [3]. The most important and serious complication of C. sinensis infection is cholangiocarcinoma, and the parasite has been classified as a group 1 biological carcinogen [6]. The specific diagnosis of C. sinensis is important for successful treatment and control of the infection. Diagnosis and treatment are important to prevent serious complications and, the simple and reliable diagnostic method is necessary to control clonorchiasis in endemic areas, where resources for the diagnosis are limited
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