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      Molecular Identification and Phylogenetic Analysis of Nuclear rDNA Sequences of Clonorchis sinensis Isolates From Human Fecal Samples in Heilongjiang Province, China

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          Abstract

          Studying the genetic diversity of parasite is important for understanding their biogeography and molecular epidemiology, as well as for establishing disease prevention and control strategies. Clonorchis sinensis is an important foodborne parasite worldwide. However, despite its epidemiological significance, the genetic diversity of C. sinensis has not been well studied from human in northeastern China. In this study, a total of 342 fecal specimens were collected from residents living in five villages in Heilongjiang Province and analyzed for the presence of C. sinensis by PCR amplification and sequencing of the internal transcribed spacer 1 (ITS1) and ITS2 regions of nuclear ribosomal DNA. 21.64% (74/342) of fecal samples were found to be positive for C. sinensis by PCR. The sequences of the ITS1 region in 34 of the 74 samples (45.95%) matched that of MK179278, Genetic polymorphisms were observed at six nucleotide sites. The ITS2 gene sequence of 37 of the 74 samples (50%) matched that of MK179281. In conclusion, a low degree of genetic diversity between C. sinensis isolates from China and different geographical regions was found at ITS loci. Despite this conservation, sequencing of the rDNA region has provided important data that will be useful for future studies addressing the molecular evolution, biology, medical implications and ecology of C. sinensis.

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          Most cited references35

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          Clonorchiasis.

          On Aug 21, 1875, James McConnell published in The Lancet his findings from a post-mortem examination of a 20-year-old Chinese man--undertaken at the Medical College Hospital in Calcutta, India--in whom he found Clonorchis sinensis in the bile ducts. Now, exactly 140 years later, we have a sound understanding of the lifecycle of this liver fluke, including key clinical, diagnostic, and epidemiological features. Developments in the so-called -omics sciences have not only advanced our knowledge of the biology and pathology of the parasite, but also led to the discovery of new diagnostic, drug, and vaccine targets. C sinensis infection is primarily related to liver and biliary disorders, especially cholangiocarcinoma. Clonorchiasis mainly occurs in east Asia, as a result of the region's social-ecological systems and deeply rooted cultural habit of consuming raw freshwater fish. The Kato-Katz technique, applied on fresh stool samples, is the most widely used diagnostic approach. Praziquantel is the treatment of choice and has been considered for preventive chemotherapy. Tribendimidine showed good safety and therapeutic profiles in phase 2 trials and warrants further investigation. Still today, the precise distribution, the exact number of infected people, subtle morbidities and pathogenesis, and the global burden of clonorchiasis are unknown. Integrated control strategies, consisting of preventive chemotherapy; information, education, and communication; environmental management; and capacity building through intersectoral collaboration should be advocated.
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            Clonorchiasis: an update.

            H-J Rim (2005)
            Clonorchis sinensis, the Chinese or oriental liver fluke, is an important human parasite and is widely distributed in southern Korea, China (including Taiwan), Japan, northern Vietnam and the far eastern part of Russia. Clonorchiasis occurs in all parts of the world where there are Asian immigrants from endemic areas. The human and animal reservoir hosts (dogs, pigs, cats and rats) acquire the infection from the ingestion of raw fish containing infectious metacercariae. The first intermediate snail hosts are mainly species of Parafossarulus and Bithynia. Numerous species of freshwater fish serve as the second intermediate hosts of C. sinensis. Extensive studies of clonorchiasis during several decades in Japan, Korea, China and other countries have shown much progress in proving its morphological features including ultrastructure, biology, pathogenesis, epidemiology, clinical manifestations and chemotherapy. The present review deals with mainly current results obtained on the epidemiological, pathological and clinical aspects, as well as control measures in endemic areas. As for the complications of clonorchiasis, formation of calculi in the intrahepatic biliary passages is one of the most characteristic pathological features. It is sometimes accompanied by suppurative cholangitis, cholecystitis, cholangiohepatitis and ultimately can cause cholangiocarcinoma. Experimental results on the relationship to the occurrence of cholangiocarcinoma are presented. Clinical diagnosis by radiological findings including cholangiography, sonography and computerized tomography as well as magnetic resonance imaging for biliary or pancreatic ducts are outlined. Current studies on immunology and molecular biology of C. sinensis were introduced. Praziquantel is the drug of choice for clonorchiasis. The most effective regimen is 25 mg kg(-1) three times daily (total dose, 75 mg kg(-1)) administered orally at 5- to 6-h intervals over a single day. Prevention and control measures are also discussed.
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              Current status and perspectives of Clonorchis sinensis and clonorchiasis: epidemiology, pathogenesis, omics, prevention and control

              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. Electronic supplementary material The online version of this article (doi:10.1186/s40249-016-0166-1) contains supplementary material, which is available to authorized users.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                28 January 2019
                2019
                : 10
                : 26
                Affiliations
                [1] 1Department of Parasitology, Harbin Medical University , Harbin, China
                [2] 2Department of Orthopaedics, The Fourth Affiliated Hospital of Harbin Medical University , Harbin, China
                Author notes

                Edited by: Shu Hui Chen, National Institutes of Health (NIH), United States

                Reviewed by: Kin-Ming (Clement) Tsui, Weill Cornell Medicine-Qatar, Qatar; Guo-Hua Liu, Hunan Agricultural University, China

                *Correspondence: Su Han, hshshs1982@ 123456126.com

                These authors have contributed equally to this work

                This article was submitted to Infectious Diseases, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2019.00026
                6360181
                c9d27dee-cb37-4605-9f09-fb551f286817
                Copyright © 2019 Zhang, Sun, Tang, Chen and Han.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 23 May 2018
                : 09 January 2019
                Page count
                Figures: 3, Tables: 3, Equations: 0, References: 42, Pages: 8, Words: 0
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                clonorchis sinensis,molecular identification,phylogenetic analysis,its1,its2,heilongjiang province

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