• Title/Summary/Keyword: IFAT

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Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 1. Indirect fluorescent antibody test and enzyme-linked immunosorbent assay for antibody detection of Babesia gibsoni infections in dogs (Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 1. Babesia gibsoni 진단을 위한 간접형광항체법(IFAT)과 효소표지면역검사법(ELISA))

  • Suh, Myung-deuk;Shin, Yong-seung
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.583-593
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    • 1997
  • Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

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Detection of Vibrio alginolyticus in Penaeus chinensis Using an Indirect fluorescent Antibody Technique

  • Heo, Moon-Soon;Lee, Sang-Hyeon;Son, Hong-Joo;Choon, Bok-Song;Lee, Je-Hee;Yeo, In-Okyy;Lee, Jung-Jae
    • Journal of Life Science
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    • v.10 no.1
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    • pp.24-27
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    • 2000
  • For the rapid diagnosis of vibriosis in penaeid shirmp, the indirect fluorescent antibody technique(IFAT) was established to detect Vibrio alginolyticus. The titers of the antisera used for this experiment were above 1280. Vibrio alginolyticus possesses the specific antigen, and also have antigens shared with other strains. When an V. alginolyticus-infected adult shirmp was tested by IFAT, V. alginolyticus was detected mainly in the muscle tissues near the injection point and the haemolymph but only few in other tissues. This result indicates that the pathogen bacteria could be detected by IFAT. Thus, it is suggested IFAT is more convenient and sensitive method than conventional plate method for the diagnosis of induced Vibrio infection in the penaeid shrimps.

Serodiagnosis of Extraintestinal Amebiasis: Retrospective Evaluation of the Diagnostic Performance of the Bordier® ELISA Kit

  • Beyls, Nicolas;Cognet, Odile;Stahl, Jean-Paul;Rogeaux, Olivier;Pelloux, Herve
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.71-74
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    • 2018
  • Soluble antigens from an axenic culture of Entamoeba histolytica were used to develop a commercial ELISA kit to quantify anti-E. histolytica antibodies in sera of patients with extraintestinal amebiasis in non-endemic settings. The diagnostic specificity and sensitivity of the test were assessed retrospectively using 131 human serum samples with amoebic serologic status available. They were selected according to their results in immunofluorescence (IFAT) and were separated in 2 sample categories: 64 sera with positive results by IFAT and 67 with negative results by IFAT. The sensitivity and specificity of the ELISA kit were assessed at 95.0% and 94.0% compared to the IFAT. The test can be useful to exclude a potential diagnosis of amebiasis and could be used as a screening method since ELISA is an automated technique.

Development of indirect fluorescent antibody test and the prevalence of the antibody titer for Neospora caninum of domestic animal in Korea (Neospora caninum 간접형광항체진단법 개발 및 국내 가축에서의 항체가 조사)

  • Cho, Young-mi;Kang, Seung-won;Choi, Eun-jin;Jeong, Woo-seog;Yoon, Yong-dhuk;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.595-599
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    • 1998
  • The present study was undertaken to develop the kit for indirect immunofluorescence antibody test(IFAT) and to investigate the prevalence of N caninum in cattle and dogs in Korea. The neo-antigen kit of IFAT developed in our laboratory was proved diagnostic efficacy by compared with the kit of veterinary medical research and development(VMRD). A survey of N caninum infections among cow and dogs in elevn areas of southern part of country was performed using a IFAT. The infection rate of 190 nationwide cattle was 8.4%(16/190) but was 75%(45/60) at P area in Chunnam province where the cattle showed the abortion repeatedly. Any of the dogs was not N caninum -positive in the Kangwon and Kyonggi areas.

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Detection of Edwardsiella tarda, the Pathogenic Bacteria in Freshwater Fishes by Means of the Indirect Fluorescent Antibody Technique (간접형광항체법을 이용한 담수양식어의 병원균 Edwardsiella tarda의 검출)

  • 류해진;조우영;이청산;허강준
    • Korean Journal of Veterinary Service
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    • v.16 no.2
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    • pp.111-119
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    • 1993
  • In this study, we carried out the rapid diagnostic system based on indirect fluorescent anti-body technique (IFAT) for detection of bacterial diseases in cultured freshwater fishes. 1. When the fishes were tested with graded dilution of Edwardsiella tarda FPC 470 bacteria detection from ten fishes Injected with $4.1{\times}10^3$colony forming unit(CFU) /ml, all of them were detected by IFAT but only two fishes were recognizable by the culture method in the tested fishes injected with $4.1{\times}10^3$CFU /ml. 2. The bacteria E. tarda could be detected by IFAT method from 1 to 48hrs after Injection in the tissues tested such as kidney, liver and spleen of the fishes, whereas detection by culture method could be recognized from 1 to 48hrs after injection In the kidney and spleen but it was not possible from preinjection to 1 hr in the liver. 3. Thus, IFAT proved to be more useful technique than plate culture method in the diagnosis of Edwardsiellosis in the freshwater fishes.

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Comparative evaluation of indirect immunofluorescent antibody test with enzyme-linked immunosorbent assay in serodiagnosis of human neurocysticercosls (뇌낭미충증의 혈청학적 진단에 있어서 간접 형광항체 반응 및 효소연결성 면역흡착 검사의 비교 평가)

  • Eom, Gi-Seon;Jo, Seung-Yeol;Im, Han-Jong
    • Parasites, Hosts and Diseases
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    • v.26 no.1
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    • pp.27-32
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    • 1988
  • The applicability of indirect immunoftuorescent antibody test (IFAT) was compared with enzyme-linked immunosorbent assay (ELISA) in sera from 163 cases of confirmed neurocysticercosis, 101 other neurologic and parasitic diseases and 100 normal controls. As antigen, frozen sections of a Taenia solium metacestode from a human brain was used in IFAT and cystic fluid was used in ELISA. For the detection of specific IgG antibody, IFAT was equally sensitive (89.6%) and specific (85.1%) as ELISA. The antibody titers by IFAT were correspondingly increased with mean absorbance of ELISA. The corresponding rate of positivity in the two techniques was 90.8%. Except for the difficulty in detecting antibodies in cerebrospinal fluid (CSF), IFAT was concluded to be very useful for the serodiagnosis of human neurocysticercosis.

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Seroprevalence of Antibodies to Neospora caninum in Korean Native Cattle Raised in Kangwon Province (강원도 사육 한우에서 Neospora caninum에 대한 항체양성률 조사)

  • Hwang, Eui-kyung
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.283-288
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    • 2003
  • This survey was carried out to investigate the seroprevalence of antibodies to Neospora caninum in Korean native cattle (KNC) raised in Kangwon province in Korea. A total of 867 sera collected from KNC were tested for N. caninum antibodies using an indirect immunofluorescent antibody test (IFAT). One hundred and fifty five (17.9%) KNC were positive by IFAT. Seroprevalence of cows was 19.6% (44/224) and seroprevalence of boars was 17.3% (111/643). Among the seroprevalences of cattle according to the raised areas, five counties or cities, Hwacheon was 33.3% (1/3), Wonju was 30.8% (4/13), Chuncheon was 25.8% (24/93), Hongcheon 18.3% (22/120) and Wheongsung was 16.6%(104/628). It was proved that KNC raised in Kangwon provinces exposed extensively and seriously to N. caninum.

Diagnosis of Viral Infection of Pensaeide (보리새우류의 바이러스 감염증의 진단)

  • 허문수;정초록;장경립
    • Journal of Life Science
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    • v.9 no.4
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    • pp.453-459
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    • 1999
  • Baculovirus(WSBV) was isolated from infected Penaeide was collected from shrimp farm at southern sea of Korea from 1993 to 1995. The Infectious virus was purified and used for diagnosis of infected shrimp. Anti-viral serum were used for immunological detection as enzyme linked immunoabsorbent assay(ELISA) and indirect fluorescent antibody technique(IFAT). In IFAT, stomach, lymphoid organ and antenae gland of infected shrimp showed fluorescent reaction. In ELISA, tissues of spontaneously infected shrimp appeared higher O.D. values than in artificial infected shrimp. Primer set was constructed from sequence of 420bp of cloned Baculovirus(WSBV) genome. Specific band for infected shrimp was detected in Polymerase chain reaction(PCR)

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Seroprevalence of antibodies to Neospora caninum in dairy cattle raised in Kangwon province (강원도 사육 젖소의 네오스포라포자충(Neospora caninum)에 대한 항체양성률 조사)

  • Hwang, Eui-Kyung
    • Korean Journal of Veterinary Research
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    • v.50 no.1
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    • pp.19-24
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    • 2010
  • This survey was carried out to investigate the seroprevalence of antibodies to Neospora (N.) caninum in dairy cattle raised in Kangwon province in Korea. A total of 349 sera collected from dairy cattle were tested for N. caninum antibodies using an indirect immunofluorescent antibody test (IFAT). Ninety six (27.5%) dairy cattle were positive by IFAT. Seroprevalence of cows was 28.9% (91/315) and seroprevalence of boars was 14.7% (5/34). The seroprevalences of cows were increased according to the ages from 19.6% in cows less than 2 year-old to 50.0% in cows more than 7 year-old. Among the seroprevalences of cattle according to the raised areas, five counties or cities, Wheongsung was 34.6% (27/78), Wonju was 29.3% (46/157), Hongcheon was 28.9% (13/45), Chuncheon was 15.0% (3/20) and Cheolwon was 13.3% (2/15). It was proved that dairy cattle raised in Kangwon provinces exposed extensively and seriously to N. caninum.

Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교)

  • 우영대;문희주;배형준
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.141-149
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    • 2000
  • The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

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