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Application of a loop-mediated isothermal amplification (LAMP) assay targeting <i>cox1</i> gene for the detection of <i>Clonorchis sinensis</i> in human fecal samples

By S. M. Mazidur Rahman (695443), Hyun Beom Song (644078), Yan Jin (143500), Jin-Kyoung Oh (342748), Min Kyung Lim (342750), Sung-Tae Hong (112337) and Min-Ho Choi (112334)

Abstract

<div><p>Background</p><p>Clonorchiasis 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.</p><p>Methodology/Principle findings</p><p>The loop-mediated isothermal amplification (LAMP) assay has been applied for the detection of <i>Clonorchis sinensis</i> DNA. Six primers targeting eight locations on the cytochrome c oxidase subunit 1 gene of <i>C</i>. <i>sinensis</i> were designed for species-specific amplification using the LAMP assay. The LAMP assay was sensitive enough to detect as little as 100 fg of <i>C</i>. <i>sinensis</i> 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 <i>Escherichia coli</i>. 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%.</p><p>Conclusions</p><p>To detect <i>C</i>. <i>sinensis</i> 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.</p></div

Topics: Microbiology, Molecular Biology, Biotechnology, Ecology, Cancer, Infectious Diseases, Virology, cox 1 gene, cytochrome c oxidase subunit 1 gene, Clonorchis sinensis DNA, PCR, samples Background Clonorchiasis, LAMP assay, bile stone formation, CI, sinensis genomic DNA
Year: 2017
DOI identifier: 10.1371/journal.pntd.0005995
OAI identifier: oai:figshare.com:article/5481952
Provided by: FigShare
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