Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.
Adult ; Antibodies, Helminth/*blood ; Antigens, Helminth/*blood ; Echinococcosis/*diagnosis ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Sensitivity and Specificity

Specific serum IgE levels of Clonorchis sinensis in infected humans were measured by avidin-biotin ELISA, and allergens from C. sinensis were identified by immunoblot and autoradiography. Then, allergens fractionated by Sephadex G-200 gel filtration were analyzed, and cross-reactive allergenic components of C. sinensis reacted with paragonimiasis sera were revealed. Fourteen out of 15 C. sinensis egg-positives were found to be serum IgE positive (absorbance > 0.27). Of 14 IgE-reacting allergen bands visualized, major allergens of 66, 61.5, 45, 37, 28.5, 23.5 and 15.5 KD were recognized by more than 50% of the sera of infected humans. The considerable individual variations of IgE immune responses to C. sinensis allergenic components were also noticed. C. sinensis extract was separated into 5 fractions by Sephadex G-200 gel filtration. Seventy-four KD allergen was recognized in the first fraction, 50, 45, 37, 29.5 and 28.5 KD in the third, and 15.5 KD in the fourth. Cross-reactive allergens with sera of paragonimiasis cases were identified as 66, 45, 28.5, 13 and 7.5 KD.
Allergens/*immunology ; Animal ; Antibodies, Helminth/*blood ; Antigens, Helminth/*immunology ; Clonorchiasis/*immunology ; Clonorchis sinensis/*immunology ; Cross Reactions ; Human ; Immunoglobulin E/*blood ; Support, Non-U.S. Gov't

BACKGROUNDRecently congenital infection with Schistosoma japonicum (S. japonicum) has been demonstrated in pigs, rabbits, mice and dogs. We explored the rabbit as an animal model for the congenital infection of schistosomiasis japonica and assessed the effect of a congenital S. japonicum infection on the resistance of rabbit kittens to a postnatal challenge infection.

METHODSSixteen pregnant New Zealand white rabbits were infected with a single dose of S. japonicum cercariae. The exposed animals were divided into three groups according to the gestation age at the time of infection. Diagnosis of prenatally acquired S. japonicum infection in the rabbit kittens was primarily based on serological tests in combination with parasitological and histopathological findings. Congenitally infected kittens were challenged percutaneously with 100 S. japonicum cercariae to assess the effect of a congenital S. japonicum infection on kitten resistance to a postnatal challenge infection.

RESULTSThe overall prevalence of congenital infection in offspring of infected mothers was 20% (12/60). The congenital infection rate in group L (late gestation) was much higher than in group E (early gestation) and group M (mid-gestation) (P <0.05). After a postnatal challenge infection, prenatally infected kittens had a 54.66% worm reduction rate, 41.45% egg reduction rate, and 51.76% granuloma size reduction rate compared to naïve kittens.

CONCLUSIONSThis study demonstrates the possibility of congenital infection of S. japonicum in rabbits and the resistance of congenitally infected kittens to a postnatal challenge infection. These results have important implications not only for epidemiological investigations, but also in designing government control programs for schistosomiasis.

Animals ; Antibodies, Helminth ; blood ; Female ; Immunoglobulin M ; blood ; Infectious Disease Transmission, Vertical ; Male ; Pregnancy ; Rabbits ; Schistosomiasis japonica ; congenital ; immunology ; parasitology

The aim of the present study was to confirm observations on the vertical transmission of Schistosoma japonicum in the rabbit. S. japonicum-infected pregnant rabbits were used in this study. Perfusion of mother rabbits was done 9 weeks after infection in order to obtain worm burdens in relation to their initial cercarial dose. Anti-schistosoma specific IgM antibodies in serum samples collected from rabbit kittens were detected by ELISA. Our results showed that gestation period lasted the normal 29-31 days. All the exposed mother rabbits became infected with S. japonicum. Positive IgM antibody OD values were detected in 12 out of the 60 kittens examined (20.0%). In group C and A, 40.0% and 17.9% of the kitten were congenitally infected, respectively. 18.1% of the kittens born to mothers infected with a single dose of 200 cercariae per rabbit were positives; this is not significantly different from that obtained for the 600 dose group (22.2%). Three randomly selected IgM+ kittens harbored between one and two adult worms. The livers of these kittens displayed granulomatous lesions. It is concluded that congenital S. japonicum infection does occur in the rabbit and is affected by the mother stage of pregnancy and to a lesser extent by its infection load.
Antibodies, Helminth/blood ; *Disease Transmission, Vertical ; Immunoglobulin M/blood ; Schistosoma japonicum/immunology ; Schistosoma japonicum/isolation & purification ; Schistosomiasis japonica/*transmission

OBJECTIVETo discuss the optimal immunization dose by observing the immunoprotective effects of different doses of recombinant Schistosoma japonicum (Chinese strain) signaling protein 14-3-3 (rSj14-3-3).

METHODSSj14-3-3 gene was amplified by reverse transcriptase PCR (RT-PCR), subcloned into prokaryotic expression vector pET28a, then transformed into E.coli to express by inducing. Purified rSj14-3-3 was prepared through SDS polyacrylamide gel electrophoresis (SDS-PAGE), electroelution, dialysis, then BALB/c mice were divided into 5 groups and immunized in rSj14-3-3 protein followed by challenging infection (the 1st, 2nd, and 3rd groups were immunized in 50 microg, 100 microg and 300 microg antigen, respectively. The 4th, 5th groups were immunized in Freund's adjuvant and normal saline controls). After 6 weeks of challenging infection, the mice were killed and the worm and egg reduction rates were calculated. And the mice sera in different time were taken to examine the specific anti-Sj14-3-3 IgG.

RESULTSrSj14-3-3 protein was expressed successfully. After immunizing and challenging, worm reduction was found to be 28.20% in the 1st group, 43.10% in the 2nd group, 40.00% in the 3rd group, respectively. Number of eggs in liver tissue was reduced by 41.80%, 57.50%, 55.70%, respectively. Compared the results of the tested groups to the controls, the differences were of significance by t-test (worm reduction rate: t = 6.8 in the 1st group, t = 8.7 in the 2nd group, t = 7.3 in the 3rd group, P < 0.01 in all tested groups. Egg reduction rate at the group's number above: t = 11.23, t = 11.54, t = 7.99, P < 0.01 in all tested groups). As compared the results between the tested groups by chi(2), the differences were of significance between the 1st and the 2nd groups (worm reduction rate: chi(2) = 8.96, P < 0.05; egg reduction rate: chi(2) = 15.69, P < 0.05), between the 1st and the 3rd groups, the differences were also of significance (worm reduction rate: chi(2) = 6.52, P < 0.05; egg reduction rate: chi(2) = 12.52, P < 0.05). The difference was not of significance between the 2nd and the 3rd groups (worm reduction rate: chi(2) = 1.20, P > 0.05; egg reduction rate: chi(2) = 0.93, P > 0.05). In all tested groups, total anti-Sj14-3-3 specific IgG rose markedly. IgG(1) and IgG(2a) subtypes were high, but IgG(2b) and IgG(3) were near the background in four subtypes tested.

CONCLUSIONImmunoprotection of rSj14-3-3 should have some relations with immunization dose, and the protection obtained from immunizing mice by using 100 microg antigen was the best.

14-3-3 Proteins ; administration & dosage ; immunology ; Animals ; Antibodies, Helminth ; immunology ; Antibody Formation ; Antigens, Helminth ; blood ; Female ; Helminth Proteins ; immunology ; Immunoglobulin G ; blood ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; Schistosoma japonicum ; genetics ; immunology ; Signal Transduction ; Vaccination

A 15-year-old boy, who had recently arrived back from a trip to Cambodia for a missionary camp, presented with several serpiginous thread-like skin lesions that began as small papules on the left upper extremities 2 weeks before his visit to Hospital. The skin lesions were pruritic and erythematous, and had migrated to the chest and abdomen. The histopathological findings showed only lymphocytic and eosinophilic infiltrations in the dermis of the biopsied skin lesion. The patient's serum reacted strongly to the Ancylostoma caninum antigen by an ELISA method. Therefore, he was diagnosed with cutaneous larva migrans by A. caninum. After the oral administration of albendazole and ivermectin, the skin lesions resolved without recurrence. This is the first reported case of a cutaneous larva migrans caused by Ancylostoma canimum diagnosed serologically using ELISA in Korea.
Adolescent ; Ancylostoma/*immunology ; Animals ; Antibodies, Helminth/*blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Larva Migrans/*diagnosis/parasitology ; Male

A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.
Animals ; Antibodies, Helminth/diagnostic use/isolation & purification ; Antibodies, Monoclonal/diagnostic use/isolation & purification ; Antigens, Helminth/*blood/*urine ; Diagnostic Tests, Routine/*methods ; Humans ; Immunoassay/methods ; Parasitology/*methods ; *Point-of-Care Systems ; Schistosoma mansoni/immunology/*isolation & purification ; Schistosomiasis mansoni/*diagnosis ; Sensitivity and Specificity

To obtain peptides mimicking epitope of a protective McAb SSjl4 specific to Schistosoma japonicum and investigate their immuno-protection effects. A phage random 12 peptide library was screened using purified McAb SSj14, 33 clones were picked up for specificity identification by ELISA. The epitope of each positive clones were detected by the sequencing analysis technique. The antigenicity of three positive clones (P1, P2 and P11) and their mixture cock-tail were further confirmed by Western-blotting, and their protective efficiency were evaluated by mice vaccination experiment. IL-12 level between the vaccinated mice and control mice were compared. 30 positive phage clones were obtained, which represented 11 different epitopes respectively, there were a similar sequence "H-N/Q-X-S-P/F-X-X-L-A-T" among all of the epitopes. Western-blotting showed that all of the three tested clones were recognized by McAb SSj14. Significant adult worm reduction (13.84% to approximately 52.83%), liver tissue egg reduction (34.17% to approximately 65.47%) as well as fecal egg reduction (28.89% to approximately 73.78%) were observed in mice vaccinated with phages of P1, P2, P11 and mixture of three clones when compared with those of the blank control group, among them, the mice vaccinated with the mixture of phage clones got higher protection than any of the mice injected with only one kind of clone phages. At the same time, the IL-12 level in serum of vaccinated mice was found higher than those of the blank control one, this suggest that IL-12 may correlate with the protective efficiency induced by the clone phages. The study provides a new way for developing an effective vaccine against S. japonicum.
Animals ; Antibodies, Helminth ; immunology ; Antibodies, Monoclonal ; immunology ; Antigens, Helminth ; immunology ; Epitopes ; immunology ; Interleukin-12 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Mimicry ; Peptide Library ; Schistosoma japonicum ; immunology ; Schistosomiasis japonica ; immunology ; prevention & control ; Vaccination

BACKGROUNDA monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).

METHODSA phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.

RESULTSSix positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07 x 10(6) individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.

CONCLUSIONSThe results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.

Animals ; Antibodies, Helminth ; immunology ; Antibodies, Monoclonal ; immunology ; Antigens, Helminth ; blood ; Base Sequence ; Immunoglobulin Fragments ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Peptide Library ; Rabbits ; Schistosomiasis japonica ; diagnosis ; Sensitivity and Specificity ; Serologic Tests

A total 7 outbreaks of trichinellosis have occurred in Korea, mostly as a result of consumption of raw wild boar (Sus scrofa) meat. Since only 1 serological survey on wild boars had yet been performed in Korea, the present study aimed to estimate the prevalence of trichinellosis in wild boars and some species of rodents by artificial digestion and serological examinations in Yanggu-gun, Gangwon-do, the endemic area of trichinellosis. Both the wild boar and rodent muscle samples revealed no Trichinella larvae by direct examination and artificial digestion method. However, serological examinations revealed that 4 wild boar sera samples out of 118 (3.4%) were positive to Trichinella antigen. Although the recovery of Trichinella larvae ended in a failure, it is proved for the first time that the sylvatic cycle of Trichinella has been maintained in wild boars of Gangwon-do, Korea.
Animals ; Antibodies, Helminth/*blood ; Antigens, Helminth/blood ; Female ; Male ; Republic of Korea/epidemiology ; Seroepidemiologic Studies ; Sus scrofa ; Swine ; Swine Diseases/*blood/diagnosis/epidemiology/parasitology ; Trichinella/classification/genetics/immunology/*isolation & purification