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Min-ho Choi - One of the best experts on this subject based on the ideXlab platform.
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Specificity of the loop-mediated isothermal amplification (LAMP) assay for the detection of Clonorchis sinensis genomic DNA.
2017Co-Authors: S. Mazidur M. Rahman, Hyun Beom Song, Yan Jin, Min Kyung Lim, Min-ho ChoiAbstract:(A) Naked eye detection of LAMP products using SYBR Green I. A green color indicates a positive reaction, and an orange color indicates a negative reaction. (B) Fluorescence of LAMP products after using SYBR Green I followed by detection under UV light. (C) Agarose gel electrophoresis of LAMP products followed by ethidium bromide staining and detection under UV light. Values in the left are in base pairs. Lane 1, Clonorchis sinensis; lane 2, Metagonimus yokogawai; lane 3, Opisthorchis viverrini; lane 4, Fasciola gigantica; lane 5, Spirometra erinacei; lane 6, Diphyllobothrium latum; lane 7, Ascaris lumbricoides; lane 8, Ascaris suum; lane 9, Necator americanus; lane 10, Trichuris trichiura; lane 11, Cryptosporidium parvum; lane 12, Entamoeba histolytica; lane 13, Giardia lamblia; lane 14, Escherichia coli; lane 15, non-template control; M, molecular marker.
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Amplification of Clonorchis sinensis DNA in 120 stool samples by loop-mediated isothermal amplification (LAMP) according to the infection intensity presented as eggs per gram of feces (EPG).
2017Co-Authors: S. Mazidur M. Rahman, Sungtae Hong, Hyun Beom Song, Yan Jin, Min Kyung Lim, Min-ho ChoiAbstract:Amplification of Clonorchis sinensis DNA in 120 stool samples by loop-mediated isothermal amplification (LAMP) according to the infection intensity presented as eggs per gram of feces (EPG).
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Results of the loop-mediated isothermal amplification (LAMP) assay for the detection of Clonorchis sinensis DNA in 120 stool samples compared with the results diagnosed by Kato-Katz (KK) and real-time PCR.
2017Co-Authors: S. Mazidur M. Rahman, Sungtae Hong, Hyun Beom Song, Yan Jin, Min Kyung Lim, Min-ho ChoiAbstract:Results of the loop-mediated isothermal amplification (LAMP) assay for the detection of Clonorchis sinensis DNA in 120 stool samples compared with the results diagnosed by Kato-Katz (KK) and real-time PCR.
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in vitro maintenance of Clonorchis sinensis adult worms
Korean Journal of Parasitology, 2012Co-Authors: Md Hafiz Uddin, Youngmee Bae, Min-ho Choi, Sungtae HongAbstract:Clonorchis sinensis is a biological carcinogen inducing human cholangiocarcinoma, and clonorchiasis is one of the important endemic infectious diseases in East Asia. The present study investigated survival longevity of C. sinensis adult worms in various in vitro conditions to find the best way of keeping the worms longer. The worms were maintained in 0.85% NaCl, 1×PBS, 1×Locke's solution, RPMI-1640, DMEM, and IMDM media, and in 1×Locke's solution with different supplements. All of the worms died within 3 and 7 days in 0.85% NaCl and 1×PBS, respectively, but survived up to 57 days in 1×Locke's solution. The worms lived for 106 days in DMEM, and 114 days in both RPMI-1640 and IMDM media. The survival rate in RPMI-1640 medium was the highest (50%) compared to that in DMEM (20±10%) and in IMDM (33.3±25.2%) after 3 months. The 1×Locke's solution with 0.005% bovine bile supplement showed increased duration of maximum survival from 42 days to 70 days. Higher concentration of bile supplements than 0.005% or addition of glucose were disadvantageous for the worm survival. The worms died rapidly in solutions containing L-aspartic acid, L-glutamic acid, and adenine compared to L-arginine, L-serine, and L-tryptophan. In conclusion, the 1×Locke's solution best supports the worms alive among inorganic solutions for 57 days, and the RPMI-1640 medium maintains living C. sinensis adults better and longer up to 114 days in vitro than other media.
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early detection and estimation of infection burden by real time pcr in rats experimentally infected with Clonorchis sinensis
Parasitology Research, 2011Co-Authors: S. Mazidur M. Rahman, Sungtae Hong, Youngmee Bae, Min-ho ChoiAbstract: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.
Sungjong Hong - One of the best experts on this subject based on the ideXlab platform.
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Bile acids drive chemotaxis of Clonorchis sinensis juveniles to the bile duct
2018Co-Authors: Won Gi Yoo, Tae Im Kim, Jin-ho Song, Sungjong HongAbstract:Clonorchiasis is a neglected tropical disease caused by Chinese liver fluke, Clonorchis sinensis infection. C. sinensis is a biological carcinogen causing cholangiocarcinoma in humans. In the mammalian host, C. sinensis newly excysted juveniles (CsNEJs) migrate from the duodenum into the bile duct. Bile drives the chemotactic behavior of CsNEJs. Little is known about which components of bile induce the chemotaxis. We designed a chemotaxis assay panel and measured the chemotactic behavior of CsNEJs in response to bile or bile acids. The CsNEJs migrated toward 0.1–1% bile but away from 5–10% bile. The CsNEJs showed strong chemoattraction to cholic acid ≥25 mM, but chemorepulsion to lithocholic acid ≥0.25 mM. To the CsNEJs, mixture of cholic acid and lithocholic acid was chemoattractive at a ratio greater than 25:1 but chemorepulsive at one smaller than that. Regarding migration in the mammalian hosts, high concentration of lithocholic acid in the gallbladder bile may repel CsNEJs from entering it. However, bile in the hepatic bile duct has a chemoattractive strength of cholic acid but a trace amount of lithocholic acid. Collectively, our results explain why the CsNEJs migrate principally to the hepatic bile ducts, bypassing the gallbladder.
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Characterization of a novel organic solute transporter homologue from Clonorchis sinensis
2018Co-Authors: Won Gi Yoo, Jhang Ho Pak, Woonmok Sohn, Fuhong Dai, Ji-yun Lee, Sungjong HongAbstract:Clonorchis sinensis is a liver fluke that can dwell in the bile ducts of mammals. Bile acid transporters function to maintain the homeostasis of bile acids in C. sinensis, as they induce physiological changes or have harmful effects on C. sinensis survival. The organic solute transporter (OST) transports mainly bile acid and belongs to the SLC51 subfamily of solute carrier transporters. OST plays a critical role in the recirculation of bile acids in higher animals. In this study, we cloned full-length cDNA of the 480-amino acid OST from C. sinensis (CsOST). Genomic analysis revealed 11 exons and nine introns. The CsOST protein had a ‘Solute_trans_a’ domain with 67% homology to Schistosoma japonicum OST. For further analysis, the CsOST protein sequence was split into the ordered domain (CsOST-N) at the N-terminus and disordered domain (CsOST-C) at the C-terminus. The tertiary structure of each domain was built using a threading-based method and determined by manual comparison. In a phylogenetic tree, the CsOST-N domain belonged to the OSTα and CsOST-C to the OSTβ clade. These two domains were more highly conserved with the OST α- and β-subunits at the structure level than at sequence level. These findings suggested that CsOST comprised the OST α- and β-subunits. CsOST was localized in the oral and ventral suckers and in the mesenchymal tissues abundant around the intestine, vitelline glands, uterus, and testes. This study provides fundamental data for the further understanding of homologues in other flukes.
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oxidative stress mediated mouse liver lesions caused by Clonorchis sinensis infection
International Journal for Parasitology, 2016Co-Authors: Sejung Maeng, Sungjong Hong, Tae Im Kim, Tongsoo Kim, Woonmok Sohn, Hye Won Lee, Qudsia Bashir, Taejin Lee, Jhang Ho PakAbstract:Abstract Clonorchis sinensis is a high-risk pathogenic helminth that strongly provokes inflammation, epithelial hyperplasia, periductal fibrosis, and even cholangiocarcinoma in chronically infected individuals. Chronic inflammation is associated with an increased risk of various cancers due to the disruption of redox homeostasis. Accordingly, the present study was conducted to examine the time course relationship between histopathological changes and the appearance of oxidative stress markers, including lipid peroxidation, enzymes involved in lipid peroxidation, and mutagenic DNA adducts in the livers of mice infected with C. sinensis , as well as proinflammatory cytokines in infected mouse sera. Histopathological phenotypes such as bile duct epithelial hyperplasia, periductal fibrosis, edema and inflammatory infiltration increased in infected livers in a time-dependent manner. Intense immunoreactivity of lipid peroxidation products (4-hydroxy-2-nonenal; malondialdehyde), cyclooxygenase-2, 5-lipoxygenase and 8-oxo-7,8-dihydro-2′-deoxyguanosine were concomitantly observed in these injured regions. We also found elevated expressions of cyclooxygenase-2 and 5-lipoxygenase in C. sinensis excretory-secretory product-treated cholangiocarcinoma cells. Moreover, the levels of proinflammatory cytokines such as TNF-α, ILβ-1 and IL-6 were differentially upregulated in infected sera. With regard to oxidative stress-mediated carcinogenesis, our findings suggest that C. sinensis infestation may disrupt host redox homeostasis, creating a damaging environment that favors the development of advanced hepatobiliary diseases such as clonorchiasis-associated cholangiocarcinoma.
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developmental transcriptomic features of the carcinogenic liver fluke Clonorchis sinensis
PLOS Neglected Tropical Diseases, 2011Co-Authors: Won Gi Yoo, Pyo Yun Cho, Tae Im Kim, Tongsoo Kim, Shinhyeong Cho, Dae Won Kim, Sanghaeng Choi, Hongseog Park, Sungjong HongAbstract:Clonorchis sinensis is the causative agent of the life-threatening disease endemic to China, Korea, and Vietnam. It is estimated that about 15 million people are infected with this fluke. C. sinensis provokes inflammation, epithelial hyperplasia, and periductal fibrosis in bile ducts, and may cause cholangiocarcinoma in chronically infected individuals. Accumulation of a large amount of biological information about the adult stage of this liver fluke in recent years has advanced our understanding of the pathological interplay between this parasite and its hosts. However, no developmental gene expression profiles of C. sinensis have been published. In this study, we generated gene expression profiles of three developmental stages of C. sinensis by analyzing expressed sequence tags (ESTs). Complementary DNA libraries were constructed from the adult, metacercaria, and egg developmental stages of C. sinensis. A total of 52,745 ESTs were generated and assembled into 12,830 C. sinensis assembled EST sequences, and then these assemblies were further categorized into groups according to biological functions and developmental stages. Most of the genes that were differentially expressed in the different stages were consistent with the biological and physical features of the particular developmental stage; high energy metabolism, motility and reproduction genes were differentially expressed in adults, minimal metabolism and final host adaptation genes were differentially expressed in metacercariae, and embryonic genes were differentially expressed in eggs. The higher expression of glucose transporters, proteases, and antioxidant enzymes in the adults accounts for active uptake of nutrients and defense against host immune attacks. The types of ion channels present in C. sinensis are consistent with its parasitic nature and phylogenetic placement in the tree of life. We anticipate that the transcriptomic information on essential regulators of development, bile chemotaxis, and physico-metabolic pathways in C. sinensis that presented in this study will guide further studies to identify novel drug targets and diagnostic antigens.
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multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis
Parasitology Research, 2011Co-Authors: Jung Guk Shin, Sungtae Hong, Pyo Yun Cho, Tae Im Kim, Sungjong HongAbstract:Antigenic proteins from Clonorchis sinensis have been previously purified and evaluated for their antigenicity to enable the serodiagnosis of clonorchiasis. Though they were of high specificity, molecularly defined proteins were reported to be less sensitive as single antigens than crude antigen. To resolve this issue, 11 clones were selected by immunoscreening an adult C. sinensis cDNA library using infected human sera. Mixed antigens were prepared using recombinant proteins of positive clones and investigated for antigenicity by immunoblotting against C. sinensis- and helminth-infected patient sera. A mixed antigen of recombinant 28 and 26 kDa glutathion S-transferases (Cs28GST and Cs26GST) produced 76% sensitivity and 95% specificity. Furthermore, a triple mix of recombinant Cs26GST and Cs28GST with vitelline precursor protein pushed up the sensitivity to 87% and maintained specificity at 95%. It is proposed that multiple antigen mixes should be further studied to develop rapid serodiagnostic test kits for the serodiagnosis of human clonorchiasis.
Yan Huang - One of the best experts on this subject based on the ideXlab platform.
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clone expression and plasminogen binding property of three fructose 1 6 bisphosphate aldolases from Clonorchis sinensis
Tropical Biomedicine, 2020Co-Authors: Jiannan Feng, Chi Liang, Juanjuan Zhou, Zhizhi Xie, Yan HuangAbstract:Fructose-1,6-bisphosphate aldolase (FbA), a well characterized glycometabolism enzyme, has been found to participate in other important processes besides the classic catalysis. To understand the important functions of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis (CsFbAs, CsFbA-1/2/3) in host-parasite interplay, the open reading frames of CsFbAs were cloned into pET30a (+) vector and the resulting recombinant plasmids were transformed into Escherichia coli BL21 (DE3) for expression of the proteins. Purified recombinant CsFbAs proteins (rCsFbAs) were approximately 45.0 kDa on 12% SDS-PAGE and could be probed with each rat anti-rCsFbAs sera by western blotting analysis. ELISA and ligand blot overlay indicated that rCsFbAs of 45.0 kDa as well as native CsFbAs of 39.5 kDa from total worm extracts and excretory-secretory products of Clonorchis sinensis (CsESPs) could bind to human plasminogen, and the binding could be efficiently inhibited by lysine analog e-aminocaproic acid. Our results suggested that as both the components of CsESPs and the plasminogen binding proteins, three CsFbAs might be involved in preventing the formation of the blood clot so that Clonorchis sinensis could acquire enough nutrients from host tissue for their successful survival and colonization in the host. Our work will provide us with new information about the biological function of three CsFbAs and their roles in hostparasite interplay.
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Clonorchis sinensis granulin identification immunolocalization and function in promoting the metastasis of cholangiocarcinoma and hepatocellular carcinoma
Parasites & Vectors, 2017Co-Authors: Caiqin Wang, Yanli Tian, Yinjuan Wu, Tingjin Chen, Zhiyue Lv, Mei Shang, Ye Li, Lu Zhao, Xin Tang, Yan HuangAbstract:Background Long-term infections by Clonorchis sinensis are associated with cholangitis, cholecystitis, liver fibrosis, cirrhosis, and even liver cancer. Molecules from the worm play vital roles in disease progress. In the present study, we identified and explored molecular characterization of C. sinensis granulin (CsGRN), a growth factor-like protein from C. sinensis excretory/secretory products (CsESPs).
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current status and perspectives of Clonorchis sinensis and clonorchiasis epidemiology pathogenesis omics prevention and control
Infectious Diseases of Poverty, 2016Co-Authors: Zeli Tang, Yan HuangAbstract:Clonorchiasis, caused by Clonorchis sinensis (C. sinensis), is an important food-borne parasitic disease and one of the most common zoonoses. Currently, it is estimated that more than 200 million people are at risk of C. sinensis infection, and over 15 million are infected worldwide. C. sinensis infection is closely related to cholangiocarcinoma (CCA), fibrosis and other human hepatobiliary diseases; thus, clonorchiasis is a serious public health problem in endemic areas. This article reviews the current knowledge regarding the epidemiology, disease burden and treatment of clonorchiasis as well as summarizes the techniques for detecting C. sinensis infection in humans and intermediate hosts and vaccine development against clonorchiasis. Newer data regarding the pathogenesis of clonorchiasis and the genome, transcriptome and secretome of C. sinensis are collected, thus providing perspectives for future studies. These advances in research will aid the development of innovative strategies for the prevention and control of clonorchiasis.
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molecular identification immunolocalization and characterization of Clonorchis sinensis triosephosphate isomerase
Parasitology Research, 2015Co-Authors: Juanjuan Zhou, Yan Huang, Chenhui Zhou, Hua Liao, Chi LiangAbstract:Clonorchis sinensis triosephosphate isomerase (CsTIM) is a key regulatory enzyme of glycolysis and gluconeogenesis, which catalyzes the interconversion of glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. In this study, the biochemical characterizations of CsTIM have been examined. A full-length complementary DNA (cDNA; Cs105350) sequence encoding CsTIM was obtained from our C. sinensis cDNA library. The open reading frame of CsTIM contains 759 bp which encodes 252 amino acids. The amino acid sequence of CsTIM shares 60–65 % identity with other species. Western blot analysis displayed that recombinant CsTIM (rCsTIM) can be probed by anti-rCsTIM rat serum and anti-C. sinensis excretory/secretory products (anti-CsESPs) rat serum. Quantitative reverse transcription (RT)-PCR and western blotting analysis revealed that CsTIM messenger RNA (mRNA) and protein were differentially expressed in development cycle stages of the parasite, including adult worm, metacercaria, excysted metacercaria, and egg. In addition, immunolocalization assay showed that CsTIM was located in the seminal vesicle, eggs, and testicle. Moreover, rCsTIM exhibited active enzyme activity in catalytic reactions. The Michaelis constant (Km) of rCsTIM was 0.33 mM, when using glyceraldehyde 3-phosphate as the substrate. The optimal temperature and pH of CsTIM were 37 °C and 7.5–9.5, respectively. Collectively, these results suggest that CsTIM is an important protein involved in glycometabolism, and CsTIM possibly take part in many biological functions in the growth and development of C. sinensis.
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rnai mediated silencing of enolase confirms its biological importance in Clonorchis sinensis
Parasitology Research, 2014Co-Authors: Xiaoyun Wang, Yanli Tian, Wenjun Chen, Yan HuangAbstract:Clonorchis sinensis (C. sinensis) infection is still a common public health problem in freshwater fish consumption areas in Asian countries. More molecular evidence are required to speed up the prevention strategies to control this kind of infectious disease. In the present study, to confirm the biological importance of Csenolase followed by our previous observations of the key metabolic enzyme, we explored the RNA silence effect of the Csenolase-derived RNA interference (RNAi) in C. sinensis. The extramembranous region aa105–226 was selected as the target sequence of RNA silence. Csenolase-derived double strand RNA (dsRNA-Csenolase, 366 bp) was synthetized and delivered into C. sinensis by soaking approach. The penetration of dsRNA into adult worms and metacercariae was tracked using fluorescently labeled RNA. Western blotting and qRT-PCR experiments were performed to determine dsRNA-Csenolase-silencing effect. Our results showed that, after incubating for 120 h, dsRNA-Csenolase could effectively target and downregulate the expression of Csenolase in both adult worms (P < 0.001) and metacercariae (P < 0.01), resulting in a remarkable killing effect on C. sinensis adult worms (P < 0.01). Fluorescent Cy3-labeled dsRNA was mostly deposited in the uterus and vitellarium of adult worm and in the cyst wall of metacercaria. The present study is the first report of RNAi trials in C. sinensis, allowing further applications in identifying functional genes in C. sinensis.
Sungtae Hong - One of the best experts on this subject based on the ideXlab platform.
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Amplification of Clonorchis sinensis DNA in 120 stool samples by loop-mediated isothermal amplification (LAMP) according to the infection intensity presented as eggs per gram of feces (EPG).
2017Co-Authors: S. Mazidur M. Rahman, Sungtae Hong, Hyun Beom Song, Yan Jin, Min Kyung Lim, Min-ho ChoiAbstract:Amplification of Clonorchis sinensis DNA in 120 stool samples by loop-mediated isothermal amplification (LAMP) according to the infection intensity presented as eggs per gram of feces (EPG).
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Results of the loop-mediated isothermal amplification (LAMP) assay for the detection of Clonorchis sinensis DNA in 120 stool samples compared with the results diagnosed by Kato-Katz (KK) and real-time PCR.
2017Co-Authors: S. Mazidur M. Rahman, Sungtae Hong, Hyun Beom Song, Yan Jin, Min Kyung Lim, Min-ho ChoiAbstract:Results of the loop-mediated isothermal amplification (LAMP) assay for the detection of Clonorchis sinensis DNA in 120 stool samples compared with the results diagnosed by Kato-Katz (KK) and real-time PCR.
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in vitro maintenance of Clonorchis sinensis adult worms
Korean Journal of Parasitology, 2012Co-Authors: Md Hafiz Uddin, Youngmee Bae, Min-ho Choi, Sungtae HongAbstract:Clonorchis sinensis is a biological carcinogen inducing human cholangiocarcinoma, and clonorchiasis is one of the important endemic infectious diseases in East Asia. The present study investigated survival longevity of C. sinensis adult worms in various in vitro conditions to find the best way of keeping the worms longer. The worms were maintained in 0.85% NaCl, 1×PBS, 1×Locke's solution, RPMI-1640, DMEM, and IMDM media, and in 1×Locke's solution with different supplements. All of the worms died within 3 and 7 days in 0.85% NaCl and 1×PBS, respectively, but survived up to 57 days in 1×Locke's solution. The worms lived for 106 days in DMEM, and 114 days in both RPMI-1640 and IMDM media. The survival rate in RPMI-1640 medium was the highest (50%) compared to that in DMEM (20±10%) and in IMDM (33.3±25.2%) after 3 months. The 1×Locke's solution with 0.005% bovine bile supplement showed increased duration of maximum survival from 42 days to 70 days. Higher concentration of bile supplements than 0.005% or addition of glucose were disadvantageous for the worm survival. The worms died rapidly in solutions containing L-aspartic acid, L-glutamic acid, and adenine compared to L-arginine, L-serine, and L-tryptophan. In conclusion, the 1×Locke's solution best supports the worms alive among inorganic solutions for 57 days, and the RPMI-1640 medium maintains living C. sinensis adults better and longer up to 114 days in vitro than other media.
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Clonorchis sinensis and clonorchiasis, an update
Parasitology international, 2011Co-Authors: Sungtae Hong, Yueyi FangAbstract:Clonorchis sinensis is the most common human liver fluke in East Asia. Several studies proved its carcinogenesis in humans and it was reclassified as a group 1 biological carcinogen in 2009. It is still actively transmitted in endemic areas of Korea, China, Russia, and Vietnam. Currently it is estimated that more than 200 million people are at risk of infection, 15–20 million people are infected and 1.5–2 million show symptoms or complications. Several molecules and genes of the fluke have been identified and characterized. Studies on its oncogenesis and omics-based findings have been especially encouraging. Diagnosis of its infection depends mainly on detection of eggs in feces but other methods have been developed. ELISA using crude extract antigen is now popular for its diagnosis. Diagnosis by detecting DNAs from eggs in feces has been developed using PCR, real-time PCR, and LAMP, which have been found sensitive and specific. Imaging diagnosis has been studied in depth and is widely used. Any evidence of clonorchiasis, such as eggs, DNAs, or images, may lead to recommendations of chemotherapy in endemic areas. Praziquantel is the major chemotherapeutic agent for clonorchiasis and recently tribendimidine was found effective and is now under investigation as a promising chemotherapeutic alternative. Sustainable control programs which include mass chemotherapy with praziquantel and education for prevention of re-infection may reduce its morbidity and eliminate its infections in endemic areas.
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multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis
Parasitology Research, 2011Co-Authors: Jung Guk Shin, Sungtae Hong, Pyo Yun Cho, Tae Im Kim, Sungjong HongAbstract:Antigenic proteins from Clonorchis sinensis have been previously purified and evaluated for their antigenicity to enable the serodiagnosis of clonorchiasis. Though they were of high specificity, molecularly defined proteins were reported to be less sensitive as single antigens than crude antigen. To resolve this issue, 11 clones were selected by immunoscreening an adult C. sinensis cDNA library using infected human sera. Mixed antigens were prepared using recombinant proteins of positive clones and investigated for antigenicity by immunoblotting against C. sinensis- and helminth-infected patient sera. A mixed antigen of recombinant 28 and 26 kDa glutathion S-transferases (Cs28GST and Cs26GST) produced 76% sensitivity and 95% specificity. Furthermore, a triple mix of recombinant Cs26GST and Cs28GST with vitelline precursor protein pushed up the sensitivity to 87% and maintained specificity at 95%. It is proposed that multiple antigen mixes should be further studied to develop rapid serodiagnostic test kits for the serodiagnosis of human clonorchiasis.
X Q Zhu - One of the best experts on this subject based on the ideXlab platform.
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sensitive and rapid detection of Clonorchis sinensis infection in fish by loop mediated isothermal amplification lamp
Parasitology Research, 2010Co-Authors: X Q Cai, Y H Wang, D Y Qiu, G X Liu, A Lin, J D Tang, Renli Zhang, X Q ZhuAbstract:The fish-borne clonorchiasis caused by the oriental liver fluke Clonorchis sinensis is endemic in a number of countries with over 35 million people being infected globally. Rapid and accurate detection of C. sinensis in its intermediate host fish is important for the control and prevention of clonorchiasis in areas where the disease is endemic. In the present study, we established a loop-mediated isothermal amplification (LAMP) approach for the sensitive and rapid detection of C. sinensis metacercariae in fish. The specificity and sensitivity of primers designed from the C. sinensis cathepsins B3 gene were evaluated, and specific amplification products were obtained with C. sinensis, while no amplification products were detected with DNA of related trematodes, demonstrating the specificity of the assay. The LAMP assay was proved to be 100 times more sensitive than a conventional polymerase chain reaction for detection of C. sinensis. The established LAMP assay provides a useful tool for the rapid and sensitive detection of C. sinensis in fish, which has important implications for the effective control of human clonorchiasis.