• 제목/요약/키워드: latex agglutination test

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Latex 응집반응을 이용한 동물의 톡소플라즈마병 진단용 kit 개발에 관한 연구 (Development of diagnostic kit(Test-MT) for the microplate latex agglutination test of toxoplasmosis in animal)

  • 서명득;주후돈;데이빗 마-스
    • 대한수의학회지
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    • 제35권3호
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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.

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Latex 응집반응을 이용한 동물의 톡소플라즈마병 진단액 개발에 관한 연구 (Development of antigen for the microplate latex agglutination test on toxoplasmosis in animals)

  • 서명득;이응구
    • 대한수의학회지
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    • 제33권4호
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    • pp.623-632
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    • 1993
  • This study was conducted to develop a sensitized latex-antigen for serodiagnosis of toxoplasmosis in animals. Tachyzoites of T gondii(RH-strain) harvested from mouse peritoneal cavity were purified through the filtraton of polycarbonate membrane(pore size, $3.0{{\mu}m}$, Costar Co.) and disrupted by ultrasonicator. The tachyzoite suspension was ultracentrifuged for 30 min at $60,000{\times}g(4{^{\circ}C})$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $0.8{{\mu}m}$ in diameter(Sigma) were used for the preparation of sensitized latex-antigen suspension. The several parameters including the preparation conditions, incubation buffer. serum dilution buffer and stability of agglutination reactions were evaluated and the results obtained were summarized as follows : 1. The antigen consisting of a water-lysate of T gondii tachyzoites was adsorbed onto polystyrene latex particles of $0.8{{\mu}m}$ in diameter by adding a latex suspension to an equal volume of diluted antigen solution and by incubating the mixture at $37{^{\circ}C}$ under different conditions. 2. The optimum incubation buffer used for the antigen sensitization was 0.1M Tris-HCl buffer(pH 8.0). 3. The optimum serum dilution buffer used for the latex agglutination test was 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300 mM NaCl. But 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300-600 mM NaCl, 0.5% BSA and 0.01% Tween-20 improved the agglutination pattems and cleared the background of microplate well without the effects on L.A titer. 4. The time required for antigen sensitization was 40 and 60 min in incubation buffer(pH 8.0) at $37{^{\circ}C}$. But the optimun time for antigen sensitization was min at $37{^{\circ}C}$. 5. The optimun quantity of antigen absorbed on latex particles for proper agglutination was the range of 20 to $32{\mu}g$ of latex particles. 6. The optimun concentration of the latex-antigen suspension for the proper agglutination reaction was determined as 0.2%(w/v). 7. The specificity, rapidity and simplicity of the latex-particle agglutination test suggested that it might be adaptable to large scale serum screening.

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Latex 응집반응 및 polymerase chain reaction을 이용한 돼지톡소플라즈마병 감염실태 조사 (Investigation of swine toxoplasmosis by Latex agglutination and polymerase chain reaction(PCR))

  • 심항섭;최경묵;전오숙;이수진;우종태;노기완
    • 한국동물위생학회지
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    • 제31권1호
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    • pp.87-91
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    • 2008
  • Between March and October 2007, a total of 516 blood samples from pigs in the Gyeonggi province were examined for seroprevalence against toxoplasmosis by latex agglutination test (LAT) and the detection of antigenic particles among seropositive samples by PCR. In the LAT, 118 (22.8%) were positive, and the unadjusted percentage of seroprevalence rates of breeding and fattening pigs were significant difference. Positive rate (14.1%) in the breeding pigs was much lower than that (27.8%) of the fattening pigs (p<0.001, Pearson's Chi-square test). The antibody detection rate of sows was lower than fattening pigs, i.e., 15.8% (25/158) and 26% (93/358), respectively (P=0.011, Pearson's Chi-square test). Among 118 seropositive samples by LAT, 68 (57.6%) were positive in PCR for the detection of the toxoplasma specific-DNA. There was a statistical difference in the positive PCR reaction between the raising pigs(63/93 67.7%) and sows (5/25, 20%) (P<0.01).

도축우 및 도축돈에서의 Toxoplasma gondii에 대한 항체조사 (A Studies on Toxoplasmosis Antibody from Slaughtered Pigs and Cattle)

  • 정기수;안상철;김진옥;김년수;장국현
    • 한국동물위생학회지
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    • 제17권1호
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    • pp.32-36
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    • 1994
  • This study was carried out to investigate antibodies against Toxoplasma gondii by Latex agglutination test from 101 slaugutered pigs and 109 cattle. In the species, 5(4.6%) out of 109 cattle and 4(4.0%) out of 101 pigs were seropositive for Toxoplasma gondii.

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경남지역에서의 Latex응집반응을 이용한 소 톡소플라즈마 항체분포 조사 (Server on the Distribution of Bovine Toxoplasma Antubodies by Latex Agglutination Test in Gyeongnam District)

  • 이병훈;류중운;황보원;변유성;조광제
    • 한국동물위생학회지
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    • 제18권3호
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    • pp.36-41
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    • 1995
  • This study was conducted to determine the serum antibodies against toxoplasma in bovine from gyeongnam district by latex agglutination(LA) test. LA test was carried out with Toxo-MT kit(Eikon chemical co). The result obtained were summerized as follow: 1. Positive rates of toxoplasma antibodies in 1,488 bovine sera was 5.0%(75 cases) by LA test. 2. The toxoplasma antibody detection rates against 823 korean cattle and 865 dairy cattle were 1.8%(11 cases) and 7.4%(64 cases) respectively. 3. In LA test serum antibody titers in 75 positive sera were shown as 54 cases(72.0% in 1:32, 9(12.0%) in 1:64, 7(9.4%) in 1:128, 3(3.40%) in 1:256, 1(1.3%) in 512 and 1(1.3%) in 1:2,048 respectively. 4. Positive rates of toxoplasma antibodies in cattls sera from each area were 0.2∼22.0%.

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세균성 뇌수막염에서 뇌척수액 Latex 응집 추적검사의 유용성 (Prognostic Value of Repeated Cerebrospinal Latex Agglutination Testing in Bacterial Meningitis)

  • 이수정;안소현;조수진;김혜순;이승주
    • Clinical and Experimental Pediatrics
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    • 제46권4호
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    • pp.345-350
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    • 2003
  • 목 적 : 세균성 뇌수막염은 높은 사망률과 후유증을 가져오는 심각한 소아에서의 감염성 질환 중 하나이다. 본 연구에서는 이미 알려진 뇌척수액 latex 응집검사의 진단적 가치 외에도 추적검사에서 예후 예측 인자로서 유용성을 알아보고자 하였다. 방 법 : 1997년 1월부터 2002년 6월까지 세균성 뇌수막염으로 진단받고 이화의대 목동병원에서 입원치료 받았던 25명의 환아들중 뇌척수액 세균배양검사와 latex 응집검사가 모두 양성이었던 19명을 대상으로 하였다. 의무기록에서 임상증상, 뇌척수액 소견, 방사선학적 소견 그리고 신경학적 후유증에 대해 조사하였다. 뇌척수액 추적검사는 항생제 치료 후 3, 7, 14, 21일 그리고 28일에 시행하였고, 3일째 음성이 된 환아들을 I군으로, 3일 이후에도 지속적으로 양성으로 남아있는 환아들을 II군으로 나누었다. 결 과 : 대상 환아 19명의 평균 나이는 $10.6{\pm}12.3$개월(2-33개월)이었고, 원인균으로는 H. influenzae type b 11명(57.9%), Group B Streptococcus 4명(21.1%), S. pneumoniae 3명(15.7%), 그리고 E. coli 1명(5.3%)이였다. I군은 10명(52.6%), II군은 9명(47.4%)이였다. 뇌척수액 추적검사에서 세균배양검사는 3일째 모두 음성화되었고, latex 응집 추적검사는 3일째 9명(47.4%)에서 양성이었고, 7일째 31.6%, 14일째 26.3%, 21일째 10.5%에서 양성이었고, 28일째에 모두 음성이 되었다. I군에서는 2명(20.0%)에서 경막하삼출 소견을 보였다. II군에서는 9명(100.0%) 모두에서 경막하 삼출을 보였고, 이 중에서 44.4%에서 뇌수종 또는 뇌연화 소견을 보였다. 신경학적 후유증은 II군에서 감각신경성난청 1명, 경련성 질환 1명 그리고 우측반신마비 1명이 발생했고, I군에서는 발생하지 않았다. 결 론 : 세균성 뇌수막염 치료 시 뇌척수액 latex 응집 추적검사를 실시하는 것은 치료 초기에 신경학적 예후를 예측하는데 유용하였다.

ELISA 및 간접 latex 응집반응검사에 의한 임산부의 항 톡소포자충 항체가 (Toxoplasma antibody titers by ELISA and indirect latex agglutination test in pregnant women)

  • 류재숙;민득영
    • Parasites, Hosts and Diseases
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    • 제34권4호
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    • pp.233-238
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    • 1996
  • 이 연구에서는 경기도 양평군 및 광주군에 사는 임산부 899명을 대상으로 IgG-ELISA와 간접 latex 응집반응검사를 시행하여 톡소포자충에 대한 항체가를 측정하였다. IgG-ELISA에서는 0.25 이상을 양성기준으로 하였을 때 음성대조군 218명 중 4명이 양성(1.8%)인 반면 임산부에서는 39 명이 양성으로 검출되어 4.3%의 양성율을 보였다. 간접 latex 응집반응검사는 수의과학연구소에서 만든 킷트(LAT)를 사용하였는데 1:64 희석배수 이상을 양성으로 하였을 때 음성대조군은 모두 음성반응을 보였고 임산부에서는 7명(0.8%)이 양성을 보였다. 임산부중에서 1.8 이상의 반응을 보인 80명을 대상으로 일본제품인 Toxotest-MF를 적용시키고 1:32 이상을 양성의 기준으로 하였을 때 임산부 8명에서 양성반응을 보였다. LAT와 Toxotest-MT의 두 반응간의 일치율은 0.94(${\kappa}-index$ = 0.632. p < 0.01)로 높은 일치율(fair to good agreement)을 보였으므로 LAT는 톡소포자충증의 예비진단에 이용될 수 있을 것으로 생각된다.

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강남성모병원과 제주의료원 내원 환자의 간접 Latex 응집 반응에 의한 Toxoplasma 항체가 (Toxoplasma antibody titers by indirect latex agglutination test in patients of Kangnam St. Mary's Hospital and Cheju Medical Center)

  • 최원영;남호우
    • Parasites, Hosts and Diseases
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    • 제27권3호
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    • pp.171-176
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    • 1989
  • 강남성모병원의 일반 내원 환자 1,019명, 내원 환자 중 천식으로 진단된 1,030명, 그리고 제주의료원에 내원 한 환자 780면 등 총 2,829명에 대하여 Toxoplasma 항체가를 측정하기 위해 간접 latex 응집반응을 실시하였다. 간접 latex 응집반응의 양성 항체가를 1 : 32로 볼 때, 강남성모병원 일반 내원 환자에서는 19명이 양성으로 1.86%(남자 550명 중 12면으로 2.18%, 여자 469명 중 7명으로 1.49%), 천식 혹은 알레르기 환자에서는 11명이 양성으로 1.07%(남자 470명 중 4명으로 0.85%, 여자 560명 중 7명으로 1.25%)이었으며, 제주의료원 내원환자 에서는 45명으로 5.77%(남자 530명 중 33명으로 6.23%, 여자 250명 중 12명으로 4.80%)이었다. 연령적 분포로는 강남성모병원 일반 환자와 제주의료원 내원 환자의 남자에서 50대에 높은 빈도를 보였고, 전체 여자에서는 일반적으로 30대, 40대 및 60대에서 높은 빈도를 보였다. 양성 항체가를 보인 75명의 항체가별 분포로는 1 32가 32명으로 42.7%, 1 : 64가 21명으로 28.0%, 1 : 128이 8명으로 10.7%, 1 256이 6명으로 8.0%, 1 : 512가 3명으로 4.0%, 그리고 1 : 2,048 혹은 그 이상의 항체가를 보유한 사람은 5명으로 6.7%였다.

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경남 중부지역에서의 Latex응집반응을 이용한 돼지 톡스플라즈마병 항체분포 조사 (Survey on the Distrributions of Swine Toxoplasma Antibodies by Latex Agglutination Test in Gyeongnam Central Area)

  • 이병훈;황보훈;변유성;이순선;김차용;서명득
    • 한국동물위생학회지
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    • 제15권2호
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    • pp.174-183
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    • 1992
  • This study was conducted to determine the serum antibodies against toxoplasma in swine from breeding-pig farm, pig farm and abattoir by latex agglutination(LA) test. LA test was carried out with commercial Toxo-MT kit (Eiken chemical co.). The results obtained were summerized as follows : 1. The cut-off titer of positive and negative reactions by Toxo-MT antigen used in this experiment was determined as the serum dilution of 1 ; 32. 2. positive rates of toxoplasma antibodies in 823 swine sera were 17.0%(140 cases) by LA test. 3. The toxoplasma antibody detection rates against 194 swine sera in breeding-pig farm, 273 swine sera in pig farm and 356 swine in abattoir were 46.9%(91 cases), 8.4%(23 cases) and 7.3% (26 cases) , respectively. 4. In LA test serum antibody titers in 823 test sera were shown as 51 cases (36.4%) in 1 : 32, 40(28.6%) in 1;64, 17(12.1%) in 1:128, 14(10.0%) in 1:256, 10(7.1%) in 1:512, 5(3.6%) in 1:1,024, and 3(2.1%) in 1 : 2,048. 5. Positive rates of toxoplasma antibodies in swine sera from each breeding-pig farm were 20.0∼61.9%.

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Latex 응집반응에 의한 경남중부지역 돼지의 톡소플라즈마병 항체조사 (Prevalence of toxoplasma antibody in swine by microplate latex agglutination test in Kyeongnam district)

  • 이병훈;김차용;서명득
    • 대한수의학회지
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    • 제33권2호
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    • pp.263-268
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    • 1993
  • This study was conducted to detect the serum antibody of toxoplasma in swine from breeding-pig, rearing-pig farm and slaughtered pig in abattior by latex agglutination(LA) test. The perfomance of LA test was carried out with commercial Toxo-MT kit(Eiken Chemical Co.)by Tsubota and Ozawa's method. The cut-off titer of positive and negative reactions by Toxo-MT antigen used in this experiment was determined as the serum titer of 1 : 32. Positive rate of toxoplasma antibody from the total of 823 serum samples by LA test was 17.0%(140 cases). And positive rates of toxoplasma antibody against serum samples of 194 from breeding-pig farm, 273 from rearing-pig farm and 356 from abattior were 91 cases(46. 9%), 23 cases(8.4%) and 26 cases(7.3%), respectively. The distributions of serum antibody titers in 823 test sera by LA test were shown 51 cases(36.3%) in 1:32, 40(28.6%) in 1:64, 17(12.1%) in 1:128, 14(10.0%) in 1:256, 3(2.1%) in 1:512, 5(3.6%) in 1:1024 and 3(2.1%) in 1:2048. The ranges of positive rate from the sera in each group of breeding-pig farms were 20~61.9%.

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