• Korean Journal of Veterinary Service
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• v.30 no.2
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• pp.205-209
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• 2007

A protein chip based on surface plasmon resonance (SPR) imaging was developed for measuring classical swine fever virus (CSFV) antibody using a recombinant gp55 protein as an antigen. The diagnostic potential of SPR imaging for detecting antibodies to the CSFV gp55 protein was compared with that of a enzyme -linked immunosorbent assay (ELISA) using 70 pig sera. There was a strong positive correlation between the SPR imaging and ELISA (n=70, r=0.916, p<0.01). Therefore, the SPR imaging, which is a label-free and high-through put method, is expected to be a valuable tool in the serodiagnosis of CSFV.

• Korean Journal of Microbiology
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• v.29 no.6
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• pp.345-352
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• 1991

A serine proteinase of molecular weight 60 kd was purified from culture supernatant of P. aeruginosa using DEAE-Trisacryl M ion-exchange and AcA 54 gel filtration column chromatography, and the properties of serine proteinase were characterized. By means of SDS-polyacrylamide gel electrophoresis, the molecular weight of the enzyme was 55 kd. The optimal pH for the activity of purified enzyme was 7.5. The activity of the purified enzyme was completely inhibited by Di-isopropylfluorophosphate(DFP) and N-.alpha.-p-tosyl-L-lysine choloromethyl detone(TLCK) but not by other proteinase inhibitors such as E-64, pepstatin A, 1, 10-phenanthroline. The purified enzyme was capable of degrading type I and type IV collagen. Antisera obtained from hymans infected with Pseudomonas aeruginosa reacted to the purified serine proteinase in immunoblots. These results indicate that the purified enzyme is trypsin-like serine proteinase and this enzyme of P. aeruginosa may play an important role in tissue damage as a spreading factor and may be useful for serodiagnosis of Pseudomonas infections.

• Journal of fish pathology
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• v.21 no.3
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• pp.175-180
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• 2008

Lymphocystis is a viral disease of fish primarily in marine and brackishwaters. Here we report the cloning, expression, and the serological applications of the lymphocystis disease virus (LCDV) major capsid protein (MCP). The MCP gene was amplified by PCR from the genomic DNA of LCDV isolated from Schlegel's black rockfish, Sebastes schlegeli, and expressed in E. coli. Mouse antisera raised against the purified recombinant MCP (rMCP) reacted with the viral MCP in an immunofluorescence assay, indicating that this rMCP would be useful for serological studies of field samples.

• The Journal of the Korean Society for Microbiology
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• v.20 no.1
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• pp.79-89
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• 1985

It is important to discriminate between tuberculosis and tuberculosis-like disease by Mycobacteria other than tuberculosis in the serodiagnosis of tuberculosis. But because common antigens share among Mycobacteria, their antigenicities to human are similar. Therefore degree of cross-reactivity of antibody in the sera of patients with tuberculosis between M. tuberculosis and Mycobacteria other than tuberculosis should be checked to increase the specificity in the serodiagnosis of tuberculosis. The activity levels of IgG antibody in the sera of 106 patients confirmed as active pulmonary tuberculosis and 30 normal healthy control person to the pressate extract antigen (TE, BE, AE, and FE antigen) from M. tuberculosis, M. bovis, M. avium, and M. fortuitum were measured by enzyme-linked immunosorbent assay and the crossreactivity of IgG antibody with mycobacterial species was analysed. The results were as follows; 1. The activity level(O.D. at 492nm) of IgG to TE antigen in sera of patients with pulmonary tuberculosis was $0.228{\pm}0.167$ in minimal tuberculosis; moderately advanced, $0.556{\pm}0.616$; far advanced, $1.116{\pm}0.651$ and $0.315{\pm}0.245$ in miliary tuberculosis. 2. The activity level (O.D. at 492nm) of IgG to BE antigen in sera of patients with pulmonary tuberculosis was $0.190{\pm}0.162$ in minimal tuberculosis; moderately advanced, $0.337{\pm}0.361$; far advanced, $0.713[\pm}0.460$ and $0.204{\pm}0.162$ in miliary tuberculosis. 3. The activity level (O.D. at 492nm) of IgG to AE antigen in sera of patients with pulmonary tuberculosis was $0.165{\pm}0.114$ in minimal tuberculosis; moderately advanced, $0.392{\pm}0.494$; far advenced, $0.751{\pm}0.512$ and $0.233{\pm}0.191$ in miliary tuberculosis. 4. The activity level (O.D. at 492nm) of IgG to FE antigen in sera of patients with pulmonary tuberculosis was $0.280{\pm}0.227$ in minimal tuberculosis; moderately advanced, $0.460{\pm}0.564$ ; far advanced, $0.845{\pm}0.573$ and $0.257{\pm}0.103$ in miliary tuberculosis. 5. The activity level (O.D. at 492nm) of IgG in sera of healthy control person was $0.126{\pm}0.084$ to TE antigen. $0.105{\pm}0.041$ to BE antigen, $0.103{\pm}0.052$ to AE antigen, and $0.095{\pm}0.061$ to FE antigen. 6. Degree of correlation(r) in activity level of IgG between TE antigen and BE antigen was 0.905 ; between TE antigen and AE antigen, 0.760; between TE antigen and FE antigen, 0.790, and between AE antigen and FE antigen, 0.945. 7. As O.D. above 0.200 was determined positive for the serodiagnosis of pulmonary tuberculosis, the sensitivity and specificity in ELISA using TE antigen were 80% and 87% respectively, whereas in the case of using BE antigen, 66% and 100%; in the case of using AE antigen, 62% and 100%, and in the case of using FE antigen, 72% and 93%, respecitively.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2001-2006
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• 2014

The objective of this study was to investigate the diagnostic significance of EBV antibody combined detection for nasopharyngeal carcinoma (NPC) in a high incidence region of southern China. Two hundred and eleven untreated NPC patients, 203 non-NPC ENT patients, and 210 healthy controls were recruited for the study. The titers of VCA/IgA and EA/IgA were assessed by immunoenzyme assay, and the levels of Rta/IgG and EBNA1/IgA were determined by enzyme-linked immunosorbent assay. The levels of VCA/IgA, EA/IgA, Rta/IgG and EBNA1/IgA demonstrated no association with gender or age (p>0.05). The receiver operating characteristic curve and the area under the curve were used to evaluate the diagnostic value. The sensitivity of VCA/IgA (98.1%) and the specificity of EA/IgA (98.5%) were the highest. When a logistic regression model was used to combine the results from multiple antibodies to increase the accuracy, the combination of VCA/IgA+Rta/IgG, whose area under the curve was 0.99, had the highest diagnostic efficiency, and its sensitivity, specificity and Youden index were 94.8%, 98.0% and 0.93 respectively. The data suggest that the combination of VCA/IgA+Rta/IgG may be most suitable for NPC serodiagnosis.

• Parasites, Hosts and Diseases
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• v.31 no.2
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• pp.141-148
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• 1993

ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting un-n westemanl specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchlasls cases (12.5%),3 of 26 cystlcercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-Inhibition test using a R westermcni specific Mab, 100% of patagonimlasls cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-Inhlbltlon test using a Mab showed higher specificity in comparison with macro-ELISA for serodlgnosis of human paragonimlasis.

• The Journal of the Korean Society for Microbiology
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• v.21 no.2
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• pp.181-189
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• 1986

To apply ELISA to serodiagnosis of leptospirosis with killed whole cells from Leptospira interrogans serovars mwogolo (Mwogolo), copenhageni (M-20), WH-20, autumnalis (Akiyami A), cynopteri (3522 C), australis (Bacillico) and Leptospira biflexa serovar patoc (patoc 1), sensitivity and specificity was evaluated. The reactivity of IgM and IgG antibody in the sera from patients with leptospirosis, hemorrhagic fever with renal syndrome and other febrile disease and normal healthy control to the killed whole cells was analysed. The results were summarized as follows. 1. The reactivity (absorbance at 492mn) of IgM and IgG to L. mwogolo antigen in the sera of pattients with leptospirosis were $1.414{\pm}0.370$, $1.242{\pm}9.554$ respectively: hemorrhagic fever with renal syndrome, $0.329{\pm}0.131$, $0.239{\pm}0.126$; other febrile disease, $0.196{\pm}0.071$, $0355{\pm}0.141$; normal healthy control, $0.136{\pm}0.016$, $0.208{\pm}0.077$. 2. The reactivity of IgM and IgG to L. copenhageni, WH-20, L. autumnalis, L. cynopteri and L. anstralis antigens were similar to that to L. mwogolo antigen, but that to L. biflexa antigen was not discriminated among above disease. 3. Correlation coefficient between the MAT titer and ELISA OD (IgM) to the above antigens was in the range of 0.071-0.518. 4. As absorbance above 0.60 was determined positive for the diagnosis of leptospirosis, the sensitivity and specificity of IgG was 25-89% and 91-96% respectively. And those of IgM was 98-100% and 89-100% except L. biflexa (29%) respectively.

• Parasites, Hosts and Diseases
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• v.52 no.5
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• pp.493-499
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• 2014

The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of Toxoplasma gondii tachyzoites and assessed it by testing 154 positive, 100 negative, and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21, and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as the positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II $IgG^{(R)}$ kit recently recommended as a confirmatory test (96.7% of concordance).

• Parasites, Hosts and Diseases
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• v.51 no.4
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• pp.467-470
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• 2013

The present study reports a human case of cutaneous gnathostomiasis with recurrent migratory nodule and persistent eosinophilia in China. A 52-year-old woman from Henan Province, central China, presented with recurrent migratory reddish swelling and subcutaneous nodule in the left upper arm and on the back for 3 months. Blood examination showed eosinophila (21.2%), and anti-sparganum antibodies were positive. Skin biopsy of the lesion and histopathological examinations revealed dermal infiltrates of eosinophils but did not show any parasites. Thus, the patient was first diagnosed as sparganosis; however, new migratory swellings occurred after treatment with praziquantel for 3 days. On further inquiring, she recalled having eaten undercooked eels and specific antibodies to the larvae of Gnathostoma spinigerum were detected. The patient was definitely diagnosed as cutaneous gnathostomiasis caused by Gnathostoma sp. and treated with albendazole (1,000 mg/day) for 15 days, and the subsequent papule and blister developed after the treatment. After 1 month, laboratory findings indicated a reduced eosinophil count (3.3%). At her final follow-up 18 months later, the patient had no further symptoms and anti-Gnathostoma antibodies became negative. Conclusively, the present study is the first report on a human case of cutaneous gnathostomiasis in Henan Province, China, based on the past history (eating undercooked eels), clinical manifestations (migratory subcutaneous nodule and persistent eosinophilia), and a serological finding (positive for specific anti-Gnathostoma antibodies).

• Parasites, Hosts and Diseases
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• v.51 no.2
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• pp.177-182
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• 2013

The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27- (KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.