• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.359-365
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• 1998

This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

• Parasites, Hosts and Diseases
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• v.35 no.4
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• pp.291-294
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• 1997

On July and August 1997 two 15-year-old mates of a football team of Tongjin Middle School in northern Kyon99i-do, Korea were consecutively diagnosed as vivax malaria by peripheral blood smear. They had no histories of travelling abroad or drug abuse. Thry witnessed that othev mates in the tram were ill of fever in the same period. A small survey was therefore undertaken to determine whether vivax malaria was outbroken locally. A total of 57 students of the team living together in a dormitory was examined for history of fever. presence of splenomegaly, blood smear and anti-p. uiuax antibody test by immunofluorescent antibody test (IFAT) . Except for the above two patients, only one case rrvraled a marginal tiler of IFAT. No other positive findings of vivax malaria were found. In the results of this local survey. no move cases of vivax malaria were revealed except the two sporadic cases.

• The Journal of Korean Society of Virology
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• v.30 no.1
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• pp.11-18
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• 2000

Since $Hantavax^{TM}$, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after primary basic vaccination with $Hantavax^{TM}$. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees at one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result $Hantavax^{TM}$ has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.

• Journal of fish pathology
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• v.36 no.2
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• pp.389-394
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• 2023

Infectious hematopoietic necrosis virus (IHNV) is s significant viral pathogen affecting cultured rainbow trout (Oncorhynchus mykiss) in Korea. In this study, five monoclonal antibodies (mAbs) (IHNV-1, 2, 3, 4, and 5) were produced using purified IHNV. Reactivities of these mAbs were analyzed by western blot (WB), enzyme-linked immunosorbent assay (ELISA), and indirect fluorescent antibody test (IFAT). These mAbs recognized glycoprotein (69 kDa, IHNV-1), nucleocapsid protein (39 kDa, IHNV-3, 4, and 5), or phosphoprotein (27 kDa, IHNV-2) of IHNV by WB analysis. ELISA results indicated that these five mAbs were specific to IHNV without showing any cross-reactivity against other fish viruses (hirame rhabdovirus, infectious pancreatic necrosis virus, and viral hemorrhagic septicemia virus). IFAT demonstrated specific fluorescence signals of IHNV-infected epithelioma papulosum cyprini (EPC) cells, whereas no reactivity of normal EPC cells was observed. These mAbs can be very useful for immuno-diagnosis of IHNV infection.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2001.09a
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• pp.36-36
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• 2001

원충성 질병인 Neospora caninum은 소와 개를 비롯한 여러 동물에서 유ㆍ사산 및 신경증상을 유발하는 질병으로 전 세계적으로 그 피해가 증가하고 있으며, 최근국내에서도 젖소농가를 중심으로 발생보고가 있으나, 한우에 대해서는 거의 역학적조사가 되어있지 않은 바, 본 연구에서는 도축되는 한우, 유ㆍ사산 및 기형송아지를 중심으로 간접형광 항체검사법(IFAT)을 이용한 혈청학적 조사 및 면역조직화학적 방법에 의한 Neospora caninum의 동정을 실시하였다. (중략)

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1151-1160
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• 1999

This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

• Journal of agricultural medicine and community health
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• v.13 no.1
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• pp.41-59
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• 1988

In many of the helminthic infections, diagnosis is accomplished by the demonstration of the eggs or, occasionally the adult worms or their parts. Diagnosis can be made by the identification of larval stage obtained from stool or surgically extracted materials too. However some kinds of parasitic disease can not be diagnosed by above mentioned procedure alone. Brain cysticercosis, ectopic paragonimiasis, Capillaria hepatica infection in liver is a good example. In such a case, immunologic method would be helpful for the decision of physician. In this paper, immunologic tools such as indirect hemagglutination test, indirect fluorescent antibody test, circumoval precipitation test, ELISA, western blot were applied for the diagnosis of Clonorchis sinenisis, Cysticercosis and C. hepatica a infection and their efficacy was evaluated. The results obtained were as follows ; 1) In the diagnosis of clonorchiasis, ELISA revealed sensitiveity of 83.3%, but cross reaction against antibody of Paragonimus westermani and Taenia species were observed. For the identification of cross reaction and species specific band of Ag-Ab reaction, western blot was applied. 59Kd relative molecular weight and 21Kd band were identified as a Clonorchis sinensis specific band. OD values of ELISA performed with sera of 18 months after praziquantel treatment decreased to half level compared to that of before treatment. Negative conversion rate of ELISA after 18 months of treatment was 60%. 2) In the diagnosis of cysticercosis, IFAT disclosed 95.8%(23/24) of sensitivity and reaction was most strongly occurred in inner membrane. ELISA revealed 90.0% (36/40) of sensitivity, but cross reaction was observed in both technique. In western blot, 91, 63 and 21Kd Mw bands were identified as a strongly positive band. Among them 63Kd band showed positive reaction against almost all sera of cysticercosis patient. 3) Circumoval precipitation, ELISA, IFAT, showed 85.0% of sensitivity in the diagnosis of C. hepatica infection in rat. The antigenic localities were inner membrane of sectioned egg antigen on the prectipitates around the mucoid plugs which were induced by circumoval precipitation reaction. Sera from rats infected with 2000eggs were collected periodically to observe the changing patterns of antibody titers by IFAT and ELISA, which showed that high titers were detected at weeks 3 and 5, then gradually declined through weeks 9until to negatively converted at weeks 13.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.1-7
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• 2014

Plasmodium vivax reemerged in the Republic of Korea (ROK) in 1993, and is likely to continue to affect public health. The purpose of this study was to measure levels of anti-P. vivax antibodies using indirect fluorescent antibody test (IFAT) in border areas of ROK, to determine the seroprevalence of malaria (2003-2005) and to plan effective control strategies. Blood samples of the inhabitants in Gimpo-si, Paju-si, and Yeoncheon-gun (Gyeonggi-do), and Cheorwon-gun (Gangwon-do) were collected and kept in Korea Centers for Disease Control and Prevention (KCDC). Out of a total of 1,774 serum samples tested, the overall seropositivity was 0.94% (n=17). The seropositivity was the highest in Paju-si (1.9%, 7/372), followed by Gimpo-si (1.4%, 6/425), Yeoncheon-gun (0.67%, 3/451), and Cheorwon-gun (0.19%, 1/526). The annual parasite incidence (API) in these areas gradually decreased from 2003 to 2005 (1.69, 1.09, and 0.80 in 2003, 2004, and 2005, respectively). The highest API was found in Yeoncheon-gun, followed by Cheorwon-gun, Paju-si, and Gimpo-si. The API ranking in these areas did not change over the 3 years. The seropositivity of Gimpo-si showed a strong linear relationship with the API of 2005 (r=0.9983, P=0.036). Seropositivity data obtained using IFAT may be useful for understanding malaria prevalence of relevant years, predicting future transmission of malaria, and for establishing and evaluating malaria control programs in affected areas.

• Parasites, Hosts and Diseases
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• v.50 no.2
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• pp.99-102
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• 2012

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels ($AI{\leq}50$), and 76 out of 82 (92.7%) sera in chronic phase of infection showed high avidity index (AI>60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.49-57
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• 1997

We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.