• Journal of Environmental Health Sciences
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• v.45 no.5
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• pp.511-519
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• 2019

Objectives: The standard method for the enumeration of environmental Legionella is culturing, which has several disadvantages, including long incubation and poor sensitivity. The purpose of this study is to demonstrate the usefulness of real-time PCR and to improve the standard method. Methods: In 200 environmental water samples, a real-time PCR and culture were conducted to detect and quantify Legionella. Using with the results of the survey, we compared the real-time PCR with the culture. Results: Each real-time PCR assay had 100% specificity and excellent sensitivity (5 GU/reaction). In the culture, 36 samples were positive and 164 samples were negative. Based on the results of the culture, real-time PCR showed a high negative predictive value of 99%, 35 samples were true positive, 105 samples were true negative, 59 samples were false positive and one sample was a false negative. Quantitative analysis of the two methods indicated a weak linear correlation ($r^2=0.29$, $r^2=0.61$, respectively). Conclusions: Although it is difficult to directly apply quantitative analysis results of real-time PCR in the enumeration of environmental Legionella, it can be used as a complementary means of culturing to rapidly screen negative samples and to improve the accuracy of diagnosis.

• Journal of Environmental Health Sciences
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• v.46 no.6
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• pp.710-718
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• 2020

Objectives: We analyzed water in waterscape facilities to investigate contamination levels of water-borne pathogens and four test items (pH, turbidity, residual chlorine, and Escherichia coli) at facilities including play fountains, splash parks, and artificial streams from June to October in Suwon City and in the whole of Gyeonggi-do. Methods: A total of 62 waterscape facility samples were collected from 36 sites and tested for pathogenic Escherichia coli and water-borne viruses that cause hand-foot-and-mouth disease, eye disease, and acute enteritis. Results: None of the water-borne pathogens were detected in waterscape facility samples collected from across Gyeonggi-do that were for pre-inspection for facility management. However, the results of samples from Suwon collected in hot weather and during the school vacation period showed five total inconsistencies in turbidity (four cases) and Escherichia coli (one case). Three out of the four inconsistent samples in turbidity were from the same facility which operated a sand filtration system due to its locational factors close to mountains. Conclusion: We suggest that the waterscape facilities in Gyeonggi-do are managed properly in the respect of microbial contamination and water quality.

• Parasites, Hosts and Diseases
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• v.36 no.3
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• pp.183-190
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• 1998

A cDNA expression library of Clonorchis sinensis adult worm was constructed, and screened out immunologically. One clone, pBCs31, was selected ill view of its predominant reactivity with an experimentally infected rabbit serum. Recombinant C. slnensis antigen iIi 28 kDa as a if-falactosidase fusion protein produced in EscherichiG coli was identified by immunoblot analysis. The cloned gene was composed of 16 copies or a 30 base pair repeat and an additional 320 bases. The deduced amino acid seqiLence of the tandem repeat was AQPPKSGDGG. On RNA slot blot analysis, C. sinensis adult worm RNA showed a positive reaction with the cloned gene Enzyme-linked immunosorbent assay using a purified recombinant antigen of pBCs31 showed higtl specificity for diagnosis of clonorchiasis.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.171-176
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• 1986

In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay (ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: 1. The specificity (95% confidence interval in negative sera of bovine fascioliasis; Mean+$2{\times}SD$ of absorbence) of the first (MW>150,000) and the second antigens(MW 120,000) were 93.7%, but those of others including crude antigen showed 100%. 2. The sensitivity(positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6%, 87.5% and 0%, respectively, but those of others showed all 100%. 3. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8. 43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. 4. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the fourth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. 5. The wheal size for bovine infected with F. hepatica was $2.46{\pm}0.15cm$ in the intradermal test antigen(saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that. of intradermal test antigen, whereas those of the fouth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

• Journal of Plant Biotechnology
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• v.8 no.1
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• pp.9-14
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• 2006

As an alternative method to produce low cost reagents for immunodiagnosis and protect the plants from viral disease, a gene encoding a single chain variable fragment(scFv) recombinant antibody targeted to the coat protein of cucumber mosaic virus (CMV) was expressed in Nacotiana tabacum. The source of the scFv recombinant antibody gene was from spleen tissue of an immunized mouse. The gene was initially cloned into the pCANTAB5E phagemid and expressed in E. coli. In the following study, the antibody gene was subcloned into the plant expression vector, pCAMBIA-1301 and introduced into tobacco leaf tissue via Agrobacterium tumefacients mediated transformation. After transformation, 56 out of 58 plants were shown to carry the desired anti-CMV scFv gene by PCR analysis. Overall, only 12.5% of the 56 putative transgenic plants were found to express the antibody to a detectable level.

• Proceedings of the Korea Contents Association Conference
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• 2015.05a
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• pp.225-226
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• 2015

뎅기 바이러스는 전 세계적으로 빠르게 확산되고 있는 모기 매개 바이러스이다. 현재까지 4가지의 혈청형이 알려져 있는 뎅기 바이러스는 뎅기열, 뎅기 출혈열과 뎅기 쇼크증후군 등을 유발한다. 현재 뎅기 바이러스가 전 세계적으로 확산되고 있음에도 불구하고 확실한 치료제나 백신이 없기 때문에 초기 진단이 매우 중요하다. 아열대기후로 진입하는 우리나라에서도 흰줄숲모기가 자주 발견되기 때문에 이 질병으로부터 결코 자유롭지가 않다. 바이러스 감염을 진단하기 위한 진단키트를 개발하기 위해서는 타깃 유전자 부위 선정이 매우 중요하기 때문에 본 연구에서는 먼저 생명정보학을 기반으로 뎅기 바이러스만을 특이적으로 검출할 수 있는 항월결정부위를 예측하고 진단키트에 적용하고자 하였다. 4가지 유형의 뎅기 바이러스 유전자들을 찾고, ProtScale Tool 프로그램으로 친수성(hydrophilicity), 접근성(acc -essibility), 유연성(flexibility), 회전(${\beta}-turns$) 등의 특성을 분석하여 항원결정부위를 선정하였다. 이 항원결정부위를 이용하여 단일클론항체를 제작하였으며 향후 진단키트로의 적용 가능성을 확인하였다.

• International Journal of Industrial Entomology and Biomaterials
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• v.4 no.2
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• pp.169-173
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• 2002

A practical technology for silkworm disease management at farmer\\`s level was suggested and test verified for its efficacy and adaptability. The technology consisted of disinfection of rearing house and appliances, early disease diagnosis, personal and rearing hygiene and silkworm body and rearing seat disinfection. Besides, the supportive techniques associated in disease management are cited. The validation trials of the technology involving 845 farmers crops with 147,530 disease free layings (dfls) of CSR bivoltine hybrids confirmed the effectiveness of the technology which resulted an average yield of 68.98 kg cocoons/100 dfls.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2635-2640
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• 2014

Sera of cancer patients may contain antibodies that react with a unique group of autologous cellular antigens called tumor-associated antigens (TAAs). The present study aimed to determine whether a mini-array of multiple TAAs would enhance antibody detection and be a useful approach in esophageal cancer detection and diagnosis. Our mini-array of multiple TAAs consisted of eleven antigens, p53, pl6, Impl, CyclinB1, C-myc, RalA, p62, Survivin, Koc, CyclinD1 and CyclinE full-length recombinant proteins. Enzyme-linked immunosorbent assays (ELISA) were used to detect autoantibodies against eleven selected TAAs in 174 sera from patients with esophageal cancer, as well as 242 sera from normal individuals. In addition, positive results of ELISA were confirmed by Western blotting. In a parallel screening trial, with the successive addition of antigen to a final total of eleven TAAs, there was a stepwise increase in positive antibody reactions. The eleven TAAs were the best parallel combination, and the sensitivity and specificity in diagnosing esophageal cancer was 75.3% and 81.0%, respectively. The positive and negative predictive values were 74.0% and 82.0%, respectively, indicating that the parallel assay of eleven TAAs raised the diagnostic precision significantly. In addition, the levels of antibodies to seven antigens, comprising p53, Impl, C-myc, RalA, p62, Survivin, and CyclinD1, were significantly different in various stages of esophageal cancer, which showed that autoantibodies may be involved in the pathogenesis and progression of esophageal cancer. All in all, this study further supports our previous hypothesis that a combination of antibodies might acquire higher sensitivity for the diagnosis of certain types of cancer. A customized mini-array of multiple carefully-selected TAAs is able to enhance autoantibody detection in the immunodiagnosis of esophageal cancer and autoantibodies to TAAs might be reference indicators of clinical stage.

• Archives of Pharmacal Research
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• v.10 no.1
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• pp.18-24
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• 1987

Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

• Parasites, Hosts and Diseases
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• v.38 no.4
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• pp.237-244
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• 2000

Although oncospheres of Taenia saginata asiatica can develop into cysticerci in immunodeficiency, immunosuppressed, and normal mice, no detailed information on the development features of these cysticerci from SCID mice is available. In the present study, the tumor-like cyst was found in the subcutaneous tissues of each of 10 SCID mice after 38-244 days inoculation with 39,000 oncospheres of T. s. asiatica. These cysts weighed 2.0-9.6 gm and were 1.5-4.3 cm in diameter. The number of cysticerci were collected from these cysts ranged from 125 to 1,794 and the cysticercus recovery rate from 0.3% to 4.6%. All cysticerci were viable with a diameter of 1-6 mm and 9 abnormal ones each with 2 evaginated protoscoleces were also found. The mean length and width of scolex, protoscolex, and bladder were $477{\;}{\times}{\;}558,{\;}756{\;}{\times}{\;}727,{\;}and{\;}1,586{\;}{\times}{\;}1,615{\;}$\mu\textrm{m}$, respectively. The diameters of suckers and rostellum were $220{\mu\textrm{m}}{\;}and{\;}70\mu\textrm{m}$, respectively All cysticerci had two rows of rostellar hooks. These findings suggest that the SCID mouse model can be employed as a tool for long-term maintenance of the biological materials for advanced studies of immunodiagnosis, vaccine development, and evaluation of cestocidal drugs which would be most benefit for the good health of the livestocks.