• Korean Journal of Veterinary Research
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• v.9 no.2
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• pp.55-59
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• 1969

For the establishment of simpler and more reliable methods for the diagnosis of Brucellosis, diagnostic antigen for Milk Ring Test (M.R.T.) was prepared according to working document recommended by Ministry of Agriculture U.S.A., and applied in the field for the test of its's applicability. The results are summarized as follows. 1. Of fifty two dairy cattle ranches in the vicinity of Seoul, five ranges were shown positive reaction to M.R.T. 2. Even though one volume of positive milk was mixed with five to ten volumes of negative milk, the positive reactions were also observed. 3. Antigenicity of the antigen was maintained without deteriolation for 13 months at $5^{\circ}C$ and three months at both $22^{\circ}C$ and $37^{\circ}C$. 4. Three positive cows to M.R.T. were also shown suspective reaction by Serum Agglutination Test.

• Parasites, Hosts and Diseases
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• v.51 no.6
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• pp.751-754
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• 2013

Neurognathostomiasis is a severe form of human gnathostomiasis which can lead to disease and death. Diagnosis of neurognathostomiasis is made presumptively by using clinical manifestations. Immunoblotting, which recognizes antigenic components of molecular mass 21 kDa and 24 kDa in larval extracts of Gnathostoma spinigerum (Gs 21/24), has high sensitivity and specificity for diagnosis of neurognathostomiasis. However, only very small amounts of the Gs 21/24 antigens can be prepared from parasites harvested from natural or experimental animals. To overcome this problem, we recently produced a recombinant matrix metalloproteinase (rMMP) protein from G. spinigerum. In this study, we evaluated this rMMP alongside the Gs 21/24 antigens for serodiagnosis of human neurognathostomiasis. We studied sera from 40 patients from Srinagarind Hospital, Khon Kaen University, Thailand, with clinical criteria consistent with those of neurognathostomiasis, and sera from 30 healthy control adults from Thailand. All sera were tested for specific IgG antibodies against both G. spinigerum crude larval extract and rMMP protein using immunoblot analysis. The sensitivity and specificity for both antigenic preparations were all 100%. These results show that G. spinigerum rMMP protein can be used as an alternative diagnostic antigen, in place of larval extract, for serodiagnosis of neurognathostomiasis.

• Parasites, Hosts and Diseases
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• v.24 no.1
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• pp.49-54
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• 1986

To evaluate the immature eggs of Paragonimus westermani as a source of diagnostic antigen, about a million eggs which were excreted by 104 adult worms were collected; their saline extract(soluble egg antigen; PwSEA) was prepared. The specific IgG and IgM antibody levels were observed in experimental dog paragonimiasis by micro ESISA, using PwSEA as well as whole worm extract of 12 week-old P. westermani(PwWWE). The protein composition of the PwSEA was observed by disc-PAGE. The results could be summarized as follows: 1. Specific IgG antibody to PwSEA begant to increase on 8 weeks after the experimental infection; it maintained its high level until the observation period of 13 weeks. The levels of IgM antibody to PwSEA however, did not show any significant change. 2. Specific IgG antibody to PwWWE began to increase earlier from 2 weeks after the infection and continued to increase until the observation period of 13 weeks. Its level was much higher than that to PwSEA. Specific IgM antibody to PwWWE increased temporarily during 2-8 weeks after the infection. 3. By disc-PAGE, PwSEA showed 2 protein bands of very low motility. The bands of PwSEA corresponded to the frist and second bands in the electrophoretic pattern of PwWWE of the 12 week-old worms. The above results indicated that the PwSEA induced antibody production in dog paragonimiasis, but its antigenicity was weaker than PwWWE to be used as a diagnostic antigen.

• BMB Reports
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• v.38 no.6
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• pp.683-689
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• 2005

Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.

• Journal of Yeungnam Medical Science
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• v.21 no.2
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• pp.191-197
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• 2004

Background: This study was performed to reconsider the efficacy of transrectal ultrasonography (TRUS) in diagnosing prostate cancer by analyzing the results of a digital rectal examination (DRE), serum prostate-specific antigen (PSA) and a transrectal ultrasonography in patients with prostate specific antigen levels of 10 ng/ml or less. Materials and Methods: One-hundred and eighty one men with PSA levels of 10 ng/ml or less, who had a TRUS-guided tissue biopsy performed, were included in this study. The detection rate of prostate cancer was compared according to the TRUS result and the presence or absence of nodularity and the consistency of the prostate on DRE. Results: In a total 181 patients, there were 73 patients with PSA levels of 4 ng/ml or less and 4 of them had prostate cancer. Thre were 108 patients with PSA levels of 4-10 ng/ml and 18 of them were prostate cancer. TRUS was performed in 152 patients and 16 out of 58 patients diagnosed with prostate cancer, 3 out of 39 diagnosed with suspicious prostate cancer, and 2 out of 55 patients diagnosed as having no prostate cancer were found to have prostate cancer. In 40 patients, a nodule was palpated on DRE and 8 of them were found to have prostate cancer. Five out of 19 patients with a stony hard consistency, 3 of 12 with a firm to hard consisency, 12 of 129 with a firm consistency, 0 of 13 with a soft to firm consistency, and 2 of 8 with a soft consistency were prostate cancer. In the prostate cancer patients, there were 4 patients with PSA levels of 4 ng/ml or less and all these patients were diagnosed with prostate cancer or suspicious prostate cancer on TRUS but the nodule was not palpated in all patients. Two were soft and 2 were firm consistency on DRE. Conclusion: In patients with serum PSA levels of 10 ng/ml or less, TRUS is a more useful supporting method than DRE and a more active application of TRUS may lead to an early diagnosis and pertinent treatment of prostate cancer.

• Parasites, Hosts and Diseases
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• v.55 no.6
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• pp.623-630
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• 2017

Plasmodium lactate dehydrogenase (pLDH) is a strong target antigen for the determination of infection with Plasmodium species specifically. However, a more effective antibody is needed because of the low sensitivity of the current antibody in many immunological diagnostic assays. In this study, recombinant Plasmodium vivax LDH (PvLDH) was experimentally constructed and expressed as a native antigen to develop an effective P. vivax-specific monoclonal antibody (mAb). Two mAbs (2CF5 and 1G10) were tested using ELISA and immunofluorescence assays (IFA), as both demonstrated reactivity against pLDH antigen. Of the 2 antibodies, 2CF5 was not able to detect P. falciparum, suggesting that it might possess P. vivax-specificity. The detection limit for a pair of 2 mAbs-linked sandwich ELISA was 31.3 ng/ml of the recombinant antigen. The P. vivax-specific performance of mAbs-linked ELISA was confirmed by in vitro-cultured P. falciparum and P. vivax-infected patient blood samples. In conclusion, the 2 new antibodies possessed the potential to detect P. vivax and will be useful in immunoassay.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.87-94
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• 1997

Clonorchis sinearis is a liver fluke which is the most prevalent helminth of humans in Korea. The better diagnostic measure of clonorchiasis is required for its nationwide control program. The present study observed antigenic bands of C. sinensis and reacting immunoglobulins in serum of infected residents. Adult C. sinensis were recovered from experimentally infected rabbits and soluble crude extract of the worms was used as the antigen after supplementation of E-64, a cysteine proteinase inhibitor. SDS- PAGE of the crude antigen resolved more than 20 protein bands between 200 and 14 kDa. The sera of infected humans collected at an endemic village showed specific IgG and IgE antibodies but little IgM and IgA antibodies. The protein bands of 94, 80, 72, 68, 52, 47, 43, 37, 34, and 28-25 kDa strongly reacted with serum Ig(GMA) or IgG antibody and 64, 62, 52, 47,44, 34,28, and 26 kDa bands reacted with serum IgE antibody. However, the 94, 80, 72, 68, 64, 62, 52, 47, and 40 kDa bands of C. sinensis antigen were found non specific. The protein bands of 43, 34, and 28-25 kDa of C. sinensis are primary target molecules of further analysis.

• Parasites, Hosts and Diseases
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• v.50 no.3
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• pp.233-238
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• 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.

• Korean Journal of Veterinary Service
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• v.38 no.1
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• pp.37-42
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• 2015

The aim of the present study was to compare the non-invasive methods for the diagnosis of H. felis with HpSA kit-based detection method and H. felis-specific PCR assay with dog's stool samples without sacrifice. Male Beagle dogs (n=6) were infected with H. felis ATCC 49179 ($1.0{\times}10^9CFU/dog$) by intra-gastric inoculation two times at 3-day intervals, and the stool specimens of dogs were collected 1, 3, 5, 7, 14, 21 days after infection to submit to HpSA test and H. felis-specific PCR. As the results, the sensitivity of the HpSA and the PCR analysis was 50.0%, 83.3% respectively. Although HpSA test is less sensitive, it could be used for rapid, cheap and easy screening assay for H. felis infection in dog and cats. We suggest that the H. pylori stool antigen kit, HpSA, is useful and effective for monitoring H. felis infection. If HpSA test would be made with H. felis antibodies in the future, its sensitivity could be increased. Also, PCR assay could be successfully used to detect the H. felis in stools. Applying the H. pylori stool antigen kit and PCR assay may be the recommended non-invasive strategy to identify H. felis in dog and cats.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.583-593
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• 1995

The present study was conducted to develop a toxoplasma latex agglutination test antigen(Test-MT) and evaluate the toxoplasma latex agglutination(LA) test using a newly-made "Test-MT kit" by comparing with the Toxo-MT kit(Eiken chemical co, Tokyo). Also, the specifity and sensitivity test were made by comparing with IFA test and IgG-ELISA. Tachyzoite suspensions of Toxoplasma gondii(RH strain) were ultracentrifuged for 30min at $60,000{\times}g(4^{\circ}C)$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $1.0{\mu}m$ in diameter(Polyscience co) were used for the preparation of sensitized latex-antigen supension(Test-MT). The frequency distribution of LA titers in Test-MT showed two peaks at <1:32 and 1:128. The borderline titer for positive test in Test-MT was determined to be 1:64. But the frequency distribution of LA tites in Toxo-MT showed two peaks at <1:16 and 1:64. The positive borderline was determined to be 1:32. Agreement of reactions between Test-MT and Toxo-MT kit by LA test was shown 92.5% in bovine sera and 97.0% in swine sera, respectively. From the results obtained here it was determined that the sensitized latex-antigen, Test-MT kit, for the microtiter agglutination test prepared as same as by the procedure described in the previous paper(Suh and Lee, 1993) was useful as a highly specific, sensitive and stable immunotiteration reagent for serodiagnosis of toxoplasma infection in animal sera.