The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was found to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplex protein originating from various organs of adult C. sinensis, and that it is composed of several 7 and 8 kDa proteins.
Animals ; Antigens, Helminth/immunology/*isolation & purification/metabolism ; Clonorchiasis/immunology ; Clonorchis sinensis/anatomy & histology/*immunology/metabolism ; Helminth Proteins/immunology/*isolation & purification/metabolism ; Human ; Mice ; Mice, Inbred BALB C ; Molecular Weight ; Support, Non-U.S. Gov't

In the course of immunoscreening of Clonorchis sinensis cDNA library, a cDNA CsRP12 containing a tandem repeat was isolated. The cDNA CsRP12 encodes two putative peptides of open reading frames (ORFs) 1 and 2 (CsRP12-1 and -2). The repetitive region is composed of 15 repeats of 10 amino acids. Of the two putative peptides, CsRP12-1 was proline-rich and found to have homologues in several organisms. Recombinant proteins of the putative peptides were bacterially produced and purified by an affinity chromatography. Recombinant CsRP12-1 protein was recognized by sera of clonorchiasis patients and experimental rabbits, but recombinant CsRP12-2 was not. One of the putative peptide, CsRP12-1, is designated CsPRA, proline-rich antigen of C. sinensis. Both the C-termini of CsRP12-1 and -2 were bacterially produced and analysed to show no antigenicity. Recombinant CsPRA protein showed high sensitivity and specificity. In experimental rabbits, IgG antibodies to CsPRA was produced between 4 and 8 weeks after the infection and decreased thereafter over one year. These results indicate that CsPRA is equivalent to a natural protein and a useful antigenic protein for serodiagnosis of human clonorchiasis.
Amino Acid Sequence ; Animals ; Antigens, Helminth/*genetics/isolation & purification ; Base Sequence ; *Cloning, Molecular ; Clonorchis sinensis/genetics/*immunology ; DNA, Helminth ; Gene Library ; Human ; Molecular Sequence Data ; Rabbits ; Recombinant Proteins ; *Repetitive Sequences, Nucleic Acid ; Support, Non-U.S. Gov't

Seroprevalence of the IgG antibodies for Clonorchis sinensis, Paragonimus westermani, Taenia solium metacestode (cysticercus), and Spirometra erinacei plerocercoid (sparganum) was measured using enzyme-linked immunosorbent assay (ELISA) in sera of patients in Korea from 1993 to 2006. A total of 74,448 specimens referred nationwide from 121 hospitals revealed an IgG positive rate of 7.6% for the 4 parasites. The IgG positive rate (18.7%) for the 4 parasites in 1993 decreased gradually to 6.6% in 2006. Individual positive rate decreased from 5.2% (1993) to 1.6% (2006) for C. sinensis, from 2.8% (1993) to 1.1% (2006) for P. westermani, from 8.3% (1993) to 2.2% (2006) for cysticercus, and from 2.6% (1993) to 1.6% (2006) for sparganum. The positive rate was highest (21.2%) in the group of patients who ranged in age from 50-59 yr old, and in the group that was referred from the Seoul area (55.9%). In conclusion, our results suggest that tissue invading parasitic infections should always be included in differential diagnosis for patients with eosinophilia associated lesions of the central nervous system, liver, and lungs in Korea.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Animals ; Antibodies, Helminth/*blood ; Child ; Child, Preschool ; Clonorchiasis/diagnosis/*epidemiology ; Clonorchis sinensis/immunology/isolation & purification ; Cysticercosis/diagnosis/*epidemiology ; Cysticercus/immunology/isolation & purification ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Eosinophilia/immunology ; Female ; Humans ; Immunoglobulin G/blood ; Infant ; Male ; Middle Aged ; Paragonimiasis/diagnosis/*epidemiology ; Paragonimus westermani/immunology/isolation & purification ; Republic of Korea/epidemiology ; Seroepidemiologic Studies ; Sparganosis/diagnosis/*epidemiology ; Sparganum/immunology/isolation & purification

The prevalence of liver and intestinal fluke infections was determined by surveying inhabitants of Hengxuan, Fusui, and Shanglin villages which were known to be endemic for liver flukes in Guangxi, China in May 2010. A total of 718 people were examined for helminth eggs by the Kato-Katz thick smear technique, ultrasonography, immunoaffinity chromatography, and DNA sequencing. The overall egg positive rate was found to be 59.6% (28.0-70.6%) that included mixed infections with liver and intestinal flukes. Cases showing higher than 20,000 eggs per gram of feces (EPG) were detected between 1.3% and 16.2%. Ultrasonographic findings exhibited overall 28.2% (72 of 255 cases) dilatation rate of the intrahepatic bile duct. Clonorchis sinensis infection was detected serologically in 88.3% (38 of 43 cases) among C. sinensis egg positive subjects by the immunoaffinity chromatography using a specific antigen for C. sinensis. For differential diagnosis of the liver and intestinal flukes, more precise PCR and nucleotide sequencing for copro-DNA were performed for 46 egg positive cases. Mixed infections with C. sinensis and Metagonimus yokogawai were detected in 8 of 46 egg positive cases, whereas 29 specimens were positive for Haplorchis taichui. Ultrasonographic findings and immunoaffinity chromatography results showed usefulness, even in a limited way, in figuring out of the liver fluke endemicity.
Animals ; China/epidemiology ; Chromatography, Affinity ; Clonorchiasis/*epidemiology/parasitology/ultrasonography ; Clonorchis sinensis/genetics/immunology/*isolation & purification ; Coinfection ; DNA, Helminth/chemistry/genetics ; Feces/parasitology ; Female ; Heterophyidae/genetics/*isolation & purification ; Humans ; Intestines/*parasitology ; Liver/*parasitology ; Parasite Egg Count ; Polymerase Chain Reaction ; Prevalence ; Sequence Analysis, DNA ; Trematode Infections/*epidemiology/parasitology/ultrasonography

The main causes of biliary obstruction are stones and cancers. Fascioliasis is a very rare case which causes biliary obstruction. Fascioliasis is a zoonosis caused by Fasciola hepatica which infects herbivores like sheep and cattle. F. hepatica lives in the biliary system or the liver parenchyma of a host. In Korea, the occurrence of this infection in human is very rare and only few cases have been reported. A 32-year-old male presented with upper abdominal pain and jaundice. His laboratory finding revealed elevated liver transaminases. Abdomen CT scan showed mild left intrahepatic bile duct dilatation. On ERCP, adult F. hepatica worms were found and were thus removed. Concurrently, clonorchiasis was diagnosed by stool exam and serologic enzyme-linked immunosorbent assay test. Clonorchiasis was treated with praziquantel. Herein, we report a case of intrahepatic bile duct dilatation due to F. hepatica infection with concurrent Clonorchis sinensis infestation.
Adult ; Animals ; Anthelmintics/therapeutic use ; Benzimidazoles/therapeutic use ; Bile Ducts, Intrahepatic ; Cholangiopancreatography, Endoscopic Retrograde ; Clonorchiasis/complications/*diagnosis/drug therapy ; Clonorchis sinensis/immunology/isolation & purification ; Enzyme-Linked Immunosorbent Assay ; Fasciola/isolation & purification ; Fascioliasis/complications/*diagnosis/parasitology ; Humans ; Liver/enzymology ; Male ; Praziquantel/therapeutic use ; Tomography, X-Ray Computed ; Transaminases/metabolism