• Parasites, Hosts and Diseases
• /
• 제50권3호
• /
• pp.233-238
• /
• 2012

The precise diagnosis of the acute toxoplasmosis in pregnant women and immunocompromsied patients has critical importance. Most of the commercially available assays use the whole Toxoplasma soluble extract as the antigen. However, the assays currently available for the detection of specific anti-Toxoplasma antibodies may vary in their abilities to detect serum immunoglobulins, due to the lack of a purified standardized antigen. The aim of this study was production and evaluation of the usefulness of the recombinant Toxoplasma gondii GRA7 antigen for the serodiagnosis of Toxoplasma gondii IgM and IgG by ELISA. A total of 70 T. gondii IgM positive sera, 74 T. gondii IgG positive sera, and 60 sera from subjects who were not infected with T. gondii were examined. These sera were shown different absorbance values in ELISA test. To control the specificity of the rGRA7 other parasitic diseases, for example, echinococcosis, malaria, leishmaniasis, fascioliasis, and strongyloidiasis were tested of which none showed positive results. Sensitivity and specificity of the generated recombinant IgG ELISA in comparison with commercial ELISA (com ELISA) were 89% and 90%, and the sensitivity and specificity of the generated recombinant IgM ELISA were 96% and 90%, respectively. The results obtained here show that this antigen is useful for diagnostic purposes.

• 대한두경부종양학회지
• /
• 제15권1호
• /
• pp.3-8
• /
• 1999

Objective: The expression of Ki67, a proliferation marker, and p27, a cyclin dependent kinases(CDKs) inhibitor, has been studied in various human neoplasms. This study was carried out to determine whether these markers are useful in distinguishing benign from malignant lesions of the thyroid or predicting biologic behavior of malignant lesions. Material and Methods: Using immunohistochemical techniques with monoclonal antibodies to Ki67 and p27, we analyzed the expression of Ki67 and p27 in various thyroid disorders(25 follicular adenomas, 47 follicular carcinomas, 16 papillary carcinomas, 20 adenomatous goiters and 40 normal thyroid tissues). The labeling indices(LIs) were determined by counting cells expressing these markers in 1000 cells per immunostained slide. Results: Neoplastic thyroid diseases showed higher expression of Ki67 and lower expression of p27 than non-neoplastic diseases(p<0.05). The expression of p27 was significantly different between follicular adenomas($LI=55.4{\pm}5.7$) and follicular carcinomas($LI=23.2{\pm}10.2$). There was, however, no significant correlation between the degree of Ki67 and p27labeling indices and types of carcinoma or clinical aggressiveness of diseases. Conclusion: The degree of Ki67 and p27 expression was useful in distinguishing between benign from malignant thyroid lesions, particulary between follicular adenoma and follicular carcinoma, but was not directly proportional to the tumor aggressiveness.

• Parasites, Hosts and Diseases
• /
• 제39권1호
• /
• pp.57-66
• /
• 2001

In the course of immunoscreening of Clonorchis sinensis cDNA library, a cDNA CsRP12 containing a tandem repeat was isolated. The cDNA CsRP 12 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 CsRP 12-2 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 you. These results indicate that CsPRA is equivalent to a natural protein and a useful antigenic protein for serodiagnosis of human clonorchiasis.

• Parasites, Hosts and Diseases
• /
• 제47권2호
• /
• pp.153-157
• /
• 2009

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.

• Parasites, Hosts and Diseases
• /
• 제56권3호
• /
• pp.237-245
• /
• 2018

Toxoplasma gondii can infect all the vertebrates including human, and leads to serious toxoplasmosis and considerable veterinary problems. T. gondii heat shock protein 60 (HSP60) is associated with the activation of antigen presenting cells by inducing initial immune responses and releasing inflammatory cytokines. It might be a potential DNA vaccine candidate for this parasite. A pVAX-HSP60 DNA vaccine was constructed and immune responses was evaluated in Kunming mice in this study. Our data indicated that the innate and adaptive immune responses was elicited by successive immunizations with pVAX-HSP60 DNA, showing apparent increases of CD3e+CD4+ and CD3e+CD8a+ T cells in spleen tissues of the HSP60 DNA-immunized mice ($24.70{\pm}1.23%$ and $10.90{\pm}0.89%$, P<0.05) and higher levels of specific antibodies in sera. Furthermore, the survival period of the immunized mice ($10.53{\pm}4.78day$) were significantly prolonged during the acute T. gondii infection. Decrease of brain cysts was significant in the experimental group during the chronic infection (P<0.01). Taken together, TgHSP60 DNA can be as a vaccine candidate to prevent the acute and chronic T. gondii infections.

• 한국동물위생학회지
• /
• 제36권4호
• /
• pp.333-340
• /
• 2013

Canine influenza virus (CIV) is an emerging pathogen that causes severe and acute respiratory disease in dogs. In 2006, the H3N2 CIV was first identified in dogs from Guangdong province in China. The nine isolates were grouped together with the canine H3N2 viruses isolated from dogs and cats in Korea. The possible interspecies transmission of influenza A virus is very important. We carried out a serological retrospective study using invited canine serum. The hospital invited 123 dogs, first vaccination group were revealed with CIV antibody positive rate of 81.8%. the second vaccination group were detected a positive rate of 91.2%. Antibody generation rate was higher in 3~10 years dogs. Protective antibody titers were detected from 2 weeks to 12 months. thereafter below the protective antibody. The results indicate that H3N2 CIV may have been consistently circulating in dog populations. Recently. These findings showed that H3N2 CIV has the capacity to replicate in and transmit among cohoused dogs and underscore the need for continued public health surveillance. Considering the result continuous management and prevention system against CIV is required at the concentrated animal care centers. The importance of CIV surveillance in this region for understanding the genesis of this virus, and it is important to remain aware of the potential of H3N2 CIV to be transmitted from dogs to the human population.

• 대한수의학회지
• /
• 제40권1호
• /
• pp.81-85
• /
• 2000

Coxiella burnetii (C burnetii) is the causative agent of Q fever in animal and human. The distribution of the disease has been documented around world. In this study we developed the competitive enzyme linked immunosorbent assay(cELISA) and compared it with indirect immunofluorescent assay(IFA). A monoclonal antibody(Mab) against C burnetii and a peroxidase-conjugated anti-mouse IgM were used as an indicator system competing against antibody in animal serum or as an indicater of the absence of antibody. Sera were considered antibody positive when the percentage inhibition index(PI index) is upper than 30. PI index is calculated as 100-[sample OD/Mab OD)${\times}100$]. Among 162 bovine serum samples, 23 samples were antibody positive both in cELISA and IFA. And 156 samples showed same results. From goat with experimentally induced infection with C burnetii the antibody was detected 20 days early in cELISA compared to IFA. On the basis of present findings, it was demonstrated that cELISA is a reliable diagnostic method for The detection of specific antibodies against C burnetii infection.

• 대한핵의학회지
• /
• 제28권1호
• /
• pp.106-111
• /
• 1994

2-iminothiolane is known to bind $NH_2$ group of lysine in the protein and deliver SH group, which can be used to label protein with $^{99m}Tc$. In this study, we looked for the best reaction condition in which 2-iminothiolane is conjugated to human polyclonal IgG and labeling condition with $^{99m}Tc$-glucoheptonate. Labeling yield was measured with TSK G4000SW column and HPLC or precipitation with 10% TCA (trichloroacetic acid) and 1% HSA. In vivo distribution was investigated with Staphylococcal abscess bearing rats. With decreasing glucoheptonate, the labeling yield decreased. Without 2-iminothiolane, $^{99m}Tc$-glucoheptonate was bound to IgG, which seemed to be direct labeling. With increasing 2-iminothiolane upto 20 times higher than IgG, the labeling yield increased, and plateau was seen with higher molar excess of 2-iminothiolane. Polymer formation was not observed. The pH for the conjugation of 2-iminothiolane and IgG was best around 6.4. $^{99m}Tc$-2-iminothiolane-IgG showed faster blood clearance, higher renal activity and lower hepatic and splenic activity than $^{99m}Tc$-DTPA-IgG. The biodistribution of $^{99m}Tc$-2-iminothiolane-IgG with higher molar excess of 2-iminothiolane was not different from that with lower molar excess. Labeling antibodies with $^{99m}Tc$ using 2-iminothiolane can afford a possible route to simple labeling and wide clinical use of the immunoscintigraphy.

• 대한미생물학회지
• /
• 제35권4호
• /
• pp.317-324
• /
• 2000

Candida albicans is one of the most frequently isolated fungal pathogens in human. Recently, the prevalence of candida infection has markedly increased, partially due to the increase of immunocompromised hosts. Proposed virulence factors of the pathogenic Candida are the ability to form hyphae to adhere to epithelial cell surfaces, and to secrete acid proteinases and phospholipases. We measured the relative cell surface hydrophobicity (CSH) and the ability of proteinase production (PROT), phospholipase production (PLase), adherence to host epithelium (ADH), and hyphal transition (Germ). The relative risk of virulence factors was analyzed by lethality test in murine model of hematogeneously disseminated candidal infection. According to Cox's proportional hazard analysis, the statistically significant virulence factors were PROT, ADH, and CSH. PROT was the highest risk factor of them. To evaluate the applicability for the diagnosis and treatment of Candidiasis, we examined the protective effect of the active and passive immunizations with the materials purified from virulence factors and antibodies to them in Candia-infected mice model. The mean survival times of active and passive immunized groups were slightly longer than those of non-immunized groups.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제29권1호
• /
• pp.10-23
• /
• 2007

Hallmarks of clinical behaviors of adenoid cystic carcinoma(ACC) of salivary glands are the delayed onset of vascular metastasis and poor responses to classical chemotherapeutic agents. Poor prognoses from salivary ACC are caused by lung metastases that are resistant to conventional therapy. Therefore, cellular and molecular characteristics that influence the dissemination of metastatic cells are important for the design of more effective treatment of salivary ACC. Tumor angiogenesis has been known to be essential for the distant metastasis of malignant cells. So, we determined expressions of angiogenic proteins in benign (pleomorphic adenoma) and malignant (ACC, mucoepidermoid carcinoma) tumors of salivary glands and compared each other and to those in oral squamous cell carcinoma. Using surgical specimens, we performed immunohistochemical assays with anti-vascular endothelial growth factor (VEGF), VEGF receptor-2 (VEGFR-2), phosphorylated VEGFR-2 (pVEGFR-2), matrix metalloproteinase (MMP)-9, and interleukin (IL)-8 antibodies. Most angiogenic factors were overexpressed in malignant salivary tumors than in pleomorphic adenoma which is benign nature. Moreover, ACC demonstrated more expression of VEGFR-2 than that of squamous cell carcinoma which used as control. Conclusively, these data show those angiogenic factors produced by salivary gland tumors may affect the propagation and metastasis of malignant cells of salivary tumors, and could be used as biomarkers for the malignant transformation of salivary gland tumors. Prospectively, although further studies will be needed, these biomarkers related to angiogenesis can be molecular targets for the therapy of salivary ACC, which has propensity for delayed vascular metastasis.