• Title/Summary/Keyword: Latex agglutination

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

  • Suh, Myung-deuk;Lee, Eung-goo
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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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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A Detection Kit for Aeromonas hydrophila Using Antibody Sensitized Latex

  • Shin, En-Joo;Lee, Soon-Deuk;Lee, Kyung-Won;Lee, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.595-598
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    • 2000
  • Aeromonas hydrophila is a pathogen to fish as well as human. It is a food-borne disease, and causes severe mortality in fish, and sometimes severe septicemia in human. In this study, a rapid detection method using latex agglutination has been developed for A. hydrophila. Polyclonal antibodies were raised against membrane and whole cells of three isolates from rainbow trout. Among these, latex particles coated with antibodies raised against whole cells of isolate No. 2 showed the best sensitivity. With latex particles coated with this antibody, we could detect $5{\times}10^4$ CFU of A. hydrophila in 5 min. The cross-reactivity with bacteria constituting the normal intestinal microflora and other pathogens for rainbow trout was insignificant. This latex agglutination assay method produced positive reaction with all clinical isolates of A. hydrophila which were identified by species-specific PCR for 16S rRNA in A. hydrophila.

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

  • Lee, Soo Jeong;Ahn, So Hyun;Cho, Su Jin;Kim, Hae Soon;Lee, Seung Joo
    • Clinical and Experimental Pediatrics
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    • v.46 no.4
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    • pp.345-350
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    • 2003
  • Purpose : Bacterial meningitis is a serious infection of childhood associated with a significant morbidity and mortality. Repeated cerebrospinal fluid(CSF) examination is a useful prognostic indicator and a delayed sterilization is associated with a higher incidence of neurologic abnormalities. In this study we tried to determine the prognostic value of repeated CSF latex agglutination testing. Methods : We retrospectively evaluated 19 patients admitted to Ewha Womans University Mokdong Hospital for bacterial meningitis from January 1997 to June 2002. Bacterial meningitis was confirmed by a positive CSF culture and a positive CSF latex agglutination test. Repeated CSF examinations were done at three, seven, 14, 21 and 28 days after antibiotics therapy. Neuroradiologic studies were performed. Results : The mean age was $10.6{\pm}12.3months$(range; two to 33 months). The male to female ratio was 2.8 : 1. The causative organisms were Haemophilus influenzae type b 57.9%, Group B Streptococcus 21.1%, Streptococcus pneumoniae 15.7% and Escherichia coli 5.3%. Three days after the initiation of antibiotics therapy, repeated CSF latex agglutination tests persisted as positive in nine (47.4%) out of 19 cases, but all CSF cultures became negative. In those cases with negative latex agglutination tests three days after antibiotics therapy, neuroradiologic findings were completely normal. But, in cases with positive latex agglutination tests three days after antibiotics therapy, neuroradiologic abnormalities such as cerebral infarction, encephalomalasia occurred in 44.4%. Conclusion : Repeated CSF latex agglutination testing was valuable as a prognostic factor in bacterial meningitis. Neuroradiologic abnormalities may occur in cases with delayed clearance of CSF latex agglutination tests more often than in cases with negative latex agglutination tests three days after antibiotics therapy.

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

  • Suh, Myung-deuk;Joo, Hoo-don;Maass, Daivd
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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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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Investigation of swine toxoplasmosis by Latex agglutination and polymerase chain reaction(PCR) (Latex 응집반응 및 polymerase chain reaction을 이용한 돼지톡소플라즈마병 감염실태 조사)

  • Shim, Hang-Sub;Choi, Gyeong-Muk;Jeon, Oh-Sook;Lee, Su-Jin;Woo, Jong-Tae;Ro, Ki-Woan
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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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).

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

  • 정기수;안상철;김진옥;김년수;장국현
    • Korean Journal of Veterinary Service
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    • v.17 no.1
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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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Toxoplasma antibody titers by ELISA and indirect latex agglutination test in pregnant women (ELISA 및 간접 latex 응집반응검사에 의한 임산부의 항 톡소포자충 항체가)

  • 류재숙;민득영
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.233-238
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    • 1996
  • The seroepidemiologic studies on antral-ToxopIasma antibody titers were carried out using ELISA and indirect latex agglutination test. Among 899 sera prepared from pregnant women, 39 cases (4.3%) revealed positive reaction and 218 sera from middle school students showed 4 positive reaction (1.8%) by ELISA. By LAT (newly established by National Veterinary Research Institute, Korea), the sera of 7 pregnant women (0.8%) showed positive reaction. When 80 sera showing ≥ 1 :8 by LAT were used for comparing the results obtained from LAT and Toxotest-MT (Eikon Chemical Co., Japans, 7 cases and 8 sera were positive, respectively. All of 11 sera of proven toxoplasmosis patients showed positive reaction in both tests. Overall proportion of agreement between LAT kit and Toxotest-MT was 0.94 (K-index : 0.632, p < 0.01), and LAT was considered to be useful for the screening of toxoplasmosis.

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

  • 최원영;남호우
    • Parasites, Hosts and Diseases
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    • v.27 no.3
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    • pp.171-176
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    • 1989
  • Total 2,829 persons consisted of 1,019 general patients and 1,030 asthma- suspected patients who visited Kangnam 51. Mary's Hospital and 780 general patients who visited Cheju Medical Center were examined for the antibody titers of Toxo- plasma by indirect latex agglutination(ILA) test. Nineteen out of 1,019(1.86%) cases in general patients group, 11 out of 1,030 (1.07%) cases in asthma patients group, and 45 out of 780(5.77%) cases in Cheju patients group showed positive ILA titers. Concerned with the age and ILA positive cases, general and asthma patients ex- pressed more cases at thirties to sixties while Cheju patients showed high incidence at children and adolescents in addition to the above mentioned ages. Frequencies of ILA positive titers were highest in 1 : 32 and 1 : 64, and some cases showed 1 : 2,048 or higher titers.

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

  • 이병훈;류중운;황보원;변유성;조광제
    • Korean Journal of Veterinary Service
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    • v.18 no.3
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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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Clinical Usefulness of D-dimer Test with Semiquantitative Latex Agglutination Method in Pulmonary Embolism (폐색전증에서 반정량적 Latex 응집법으로 시행한 D-dimer 검사의 유용성에 관한 연구)

  • Kim, Dong Kyun;Chun, Kang Il;Kim, Yang-Ki;Lee, Young-Mok;Kim, Ki Up;Uh, Soo-taek;Kim, Yong Hoon;Park, Choon Sik;Park, No Jin;Choi, Tae Youn
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.6
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    • pp.651-655
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    • 2005
  • Background : Diagnosing a pulmonary embolism is difficult because its presenting symptoms are nonspecific and there are limitations with all of the objective tests. The D-dimer is known to be a marker of the lysis of intravascular cross-linked fibrin as a result of the activation of the endogenous fibrinolytic pathways, and the D-dimer assay is these an objective method for diagnosing a pulmonary embolism. This study assessed the benefits of the D-dimer test for diagnosing a pulmonary embolism using semiquantitative latex agglutination. Methods : The latex agglutination results of 185 patients were retrospectively reviewed. The D-dimer test was performed at the time a pulmonary embolism was suspected. Ninety patients(group I) were diagnosis with PE through spiral chest CT or a chest CT angiogram, perfusion/ventilation scans, and/or pulmonary angiogram. Ninety-five patients (group II) were found not to have a pulmonary embolism through the above tests. Results : The male to female ratio and mean age in groups I and II was 37:55, and 57 years old to 50:45 and 52 years old, respectively. When the cut off value for a positive D-dimer assay was set to $500{\mu}g$, the sensitivity, positive predictive value, negative predictive value and specificity was 86.7%, 61.4%, 79.3%, and 48.4%, respectively. Conclusion : The semiquantitative latex agglutination method in the D-dimer test has a lower sensitivity and negative predictive value than the well known ELISA test particularly for small emboli. Therefore, this test is not a suitable screening test for excluding a pulmonary embolism.