• Korean Journal of Food Science and Technology
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• v.43 no.5
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• pp.618-623
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• 2011

Escherichia coli O157:H7 has been considered as a significant food-borne pathogen since its role in causing hemorrhagic colitis and hemolytic uremic syndrome in humans was recognized. In this study, we developed an immunochromatography (ICG) assay for the detection of E. coli O157:H7. E. coli O157:H7 monoclonal antibody (EC MAb) and colloidal gold were conjugated and its specificity was determined by the ICG treated with EC MAb and antimouse IgG at test and control lines, respectively. The detection limit of the ICG was $1{\times}10^5$ CFU/mL, and no crossreactivity was observed to other E. coli strains and major food-borne pathogens. To determine the minimum enrichment time for the ICG, meats and sprouts were inoculated with $1{\times}10$ CFU/100 ${\mu}L$ of E. coli O157:H7. After enrichment time of 10 and 2 h for meats and sprouts, respectively, up to $1{\times}10$ CFU/100 ${\mu}L$ of E. coli O157:H7 could be detected by ICG.

• Clinical and Experimental Pediatrics
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• v.51 no.4
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• pp.377-382
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• 2008

Purpose : The aim of this study is to assess the usefulness of skin test by an inactivated, 1/50 diluted solution of attenuated varicella vaccine in evaluating the immune status to varicella. Methods : Total 41 subjects (22 males, 19 females, aged 1-32 years) were enrolled from July to August, 2005. Past medical history including varicella infection, varicella vaccination were investigated through questionnaires. The skin test solution was prepared from solution of attenuated varicella vaccine(Oka strain) which was inactivated by exposure to room temperature for 10 days and diluted at 1/50 with normal saline. Skin test was done by injecting 0.1 mL of the solution intradermally into the volar surface of the right forearm and sterile normal saline was used as a control on the left forearm. Positive reaction was defined when the transverse diameter of the induration was 5 mm or more. Serum varicella zoster virus specific IgG antibody test by ELISA (enzyme-linked immunosorbent assay) was done. Results : In adults, the sensitivity of the varicella zoster virus skin test compared to ELISA was 94.7% and the positive predictive value was 100%. In children, both the positive predictive value and specificity were 100% but the sensitivity and the negative predictive value were 50% and 30.7% respectively. Children showed smaller skin test reactivity compared to adults. Conclusion : The varicella zoster virus skin test using inactivated, 1/50 diluted solution of attenuated varicella vaccine was proved as one of the useful tools for evaluating the immunity and susceptibility of the varicella zoster virus.

• The Korean Journal of Nuclear Medicine
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• v.34 no.3
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• pp.243-251
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• 2000

Purpose: This prospective study was performed to evaluate the usefulness of preoperative radioimmunoscintigraphy and intraoperative scintimetric examination (radioimmunoguided surgery: RIGS) using $^{99m}Tc-anti-CEA\;F(ab')_2$ fragment. Materials and Methods: Nineteen patients with rectal cancer underwent preoperative whole body planar scintigraphy at 4 hours after injection of $^{99m}Tc-anti-CEA\;F(ab')_2$ fragment and SPECT imaging at 18 hours. Surgical operation was performed at 24 hours after injection. During laparotomy, radioactivities from intraabdominal viscera were measured by gamma probe. The radioactivities from excised tumor and lymph nodes were also measured and compared with pathology. Results: All nineteen patients were confirmed to have adenocarcinomas in the rectum. Twenty-seven of 97 excised lymph node groups had metastasis and 2 patients had liver metastasis in pathology Preoperative radioimmunoscintigraphy detected primary tumors in 11 patients (sensitivity 55%) and it could not detect any lymph nodes or liver metastasis. All patients showed high radioactivity in the kidneys, liver, spleen, and major vessels in intraoperative measurement by gamma probe, and tumor activity was not discriminated from background activity However, radioactivity from excised tumor was higher than normal rectum (T/B ratio; $3.47{\pm}2.25$). When excised lymph node activity/background activity ratio >1.5 was considered as positive criteria of metastasis, sensitivity, specificity, positive and negative predictive values were 78.6%, 73.9%, 55.0% and 89.5%, respectively. Conclusion: Radioimmunoscintigraphy using $^{99m}Tc-anti-CEA\;F(ab')_2$ has no additional value for preoperative staging and use of early RIGS using $^{99m}Tc-anti-CEA\;F(ab')_2$ is inappropriate. For early RIGS using $^{99m}Tc$ labeled antibodies in rectal cancer patients, further development of more specific antibodies and methods to reduce background activity are needed.

• Journal of Food Hygiene and Safety
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• v.18 no.3
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• pp.118-124
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• 2003

For the rapid detection of various pathogenic microorganisms from food sample, various kinds of kits have been developed and commercially available in the markets. With the advantages of speed, accuracy and easiness, the market of these kits has gradually increased for the QC and QA field of food company as well as testing facilities or laboratories. In this study, the characteristics such as the detection limit and the sensitivity of immunochromatographic type of rapid detection kit (Donga Co, Korea, D-kit) for E. coli 0157:H7 developed by monoclonal antibody were examined and also the possibility of application of the kit to food samples was evaluated. The reference kits used for comparison study were Reveal E. coli 0157:H7 (Neogen Co., USA, R-kit) and VIP EHEC kit (Biocontrol Inc., USA, V-kit) occupying major market share. In the detection limit test with the E. coli 0157:H7 reference, both R-kit and D-kit showed a distinct positive reaction in $10^4$/ml and weak positive reaction in $10^3$/ml, whereas V-kit showed a same reaction in 105/ml. Also, it was identified that the culture treated with heat showed more sensitivity than no heat treated culture. The sensitivity test was conducted against 22 isolates of E. coli 0157:H7, 7 strains of non-O157:H7 verotoxin-producing E. coli, 40 strains of E. coli with different O and H antigen type, and 38 strains of non-E. coli Enterobacteriaceae, and all of the test strains except three were showed exactly three were showed exactly the same reaction against three kinds of the tested kits. All the three kinds of kits showed a positive reaction against E. coli O157:H19, E. coli O148:H18 and Salmonella galinarium. We suppose that there might be a similarity in serological property between these three strains and O157:H7. From the test results, it can be concluded that there is (was) no difference between the D-kit developed in this study and R-kit or V-kit based on the detection limit and sensitivity.

• Tuberculosis and Respiratory Diseases
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• v.49 no.5
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• pp.558-567
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• 2000

Background : Recently, serologic techniques for tuberculosis have been developed and some of them, which are focusing on detection of serum antibodies mainly directed against specific 38-kDa Mycobacterium tuberculosis, have already been introduced into the markel. In this study, diagnostic significance of a new serologic test(ELISA kit) for pulmonary tuberculosis was evaluated. Method : Serologic test with newly developed ELISA kit was performed upon 474 individuals, who include 333 active pulmonary tuberculosis patients, 80 healthy cases, and 61 tuberculosis contact cases. This serologic test was based on the ELISA technique and designed to detect antibodies to mixed complex antigens including 38-kDa, which were developed by Erume Biotech Co., Seoul. Active pulmonary tuberculosis was diagnosed by sputum AFB smear and culture methods. Results : The seropositivities using this ELISA kit were 82.1% and 73.6% in smear-positive and negative groups among active pulmonary tuberculosis, respectively. And, it also showed that seronegativities were 97.5% and 85.2% in healthy and contact groups, respectively. As a whole, the results of our study using the ELISA kit as a diagnostic method for pulmonary tuberculosis showed 80.0% sensitivity for active pulmonary tuberculosis, 97.5% specificity, 96.1% positive predictive value, and 65.0% negative predictive value when the prevalence of tuberuclosis in the samples was 60.1%. Conclusion : Our results reveal that the detection of antibody its reaction with 38-kDa antigen of M. tuberculosis is not sufficient to be accepted as single diagnostic method for pulmonary tuberculosis. However, they suggest that ELISA kit may be considered as an adjunctive test to standard diagnostic techniques of pulmonary tuberculosis.

• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.339-353
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• 1994

Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.