scholarly journals SPECIFIC AGGLUTINATION OF TREPONEMA PALLIDUM BY SERA FROM RABBITS AND HUMAN BEINGS WITH TREPONEMAL INFECTIONS

1955 ◽  
Vol 101 (4) ◽  
pp. 367-382 ◽  
Author(s):  
Paul H. Hardy ◽  
E. Ellen Nell

A method has been described for the preparation of Treponema pallidum suspensions that are suitable for specific agglutination studies and can be stored at 4°C. for months without loss of agglutinability. Such suspensions have been shown to react with two distinct antibodies in the serum of syphilitic animals and man: Wassermann antibody and a specific treponeme agglutinin. It has been demonstrated that the agglutination of treponemes by specific treponeme agglutinin is enhanced by heat treatment or aging of the suspension, and inhibited by a divalent cation, probably Ca++, normally present in serum. This inhibition has been overcome by the use of a chelating agent, ethylene-diamine tetracetate. These findings have been utilized to devise a simple agglutination test for the diagnosis of treponeme infections that is very sensitive and highly specific. This test has been carried out with 430 human sera, and a comparison has been made of the results of the agglutination, treponemal immobilization, and standard serological tests on these sera. The agglutination test appears to have a specificity comparable to the treponemal immobilization test and considerably greater than the standard serological tests.

2002 ◽  
Vol 9 (6) ◽  
pp. 1376-1378 ◽  
Author(s):  
Arnold R. Castro ◽  
Susan E. Kikkert ◽  
Martha B. Fears ◽  
Victoria Pope

ABSTRACT Syphilitic plasma can be salvaged from discarded blood donations and converted to serum by defibrination. Sixty-nine units of plasma were treated with a stock solution of 100 U of thrombin per ml in 1 M calcium chloride and then with a 10% (wt/vol) solution of kaolin. Fibrinogen concentrations detected in initial plasma samples ranged from 94 to 4,970 mg/liter (mean, 2,532 mg/liter) for samples that were reactive by the rapid plasma reagin circle card test (RPR) and from 314 to 2,742 mg/liter (mean 1,528 mg/liter) for samples that were not reactive by the RPR. The treated samples showed no measurable fibrinogen remaining after the defibrination process. In the nontreponemal RPR for syphilis, 86% of the treated plasma samples retained the same endpoint titer as that of the initial plasma sample. When the Treponema pallidum passive-particle-agglutination test was used, 98% retained the same reactivity. In the Captia Syphilis-G enzyme immunoassay, 89% of the treated samples demonstrated no change in reactivity index, and in the fluorescent treponemal antibody absorption test, 96% showed no reduction in fluorescence. Human sera containing antibodies to syphilis are used at the Centers for Disease Control and Prevention for the preparation of reference controls or as samples for proficiency testing. Finding reactive sera is becoming more difficult due to the general decline of syphilis cases in the United States. The decreasing availability of these sera can be alleviated by salvaging plasma and converting it to serum.


2016 ◽  
Vol 10 (09) ◽  
pp. 939-946 ◽  
Author(s):  
AKM Anisur Rahman ◽  
Dirk Berkvens ◽  
Claude Saegerman ◽  
David Fretin ◽  
Noor Muhammad ◽  
...  

Introduction: This study describes the seroprevalence of human brucellosis among pyretic patients and detection of Brucella abortus DNA from seropositive pyretic patients using real-time polymerase chain reaction (rtPCR) for the first time in Bangladesh. Methodology: Blood samples were collected from 300 pyretic patients from October 2007 to May 2008 and subjected to three serological tests: Rose-Bengal plate test (RBT), standard tube agglutination test (STAT), and indirect enzyme-linked immunosorbent assay (iELISA). Risk factors were identified by multivariate Firth’s logistic regression analysis. Brucella genus (BCSP31) and species-specific (IS711) rtPCR were applied to six human sera samples. Results: The seroprevalence of brucellosis among pyretic patients was estimated to be 2.0% (95% confidence interval [CI]: 0.74–4.30). The odds of brucellosis seropositivity were 8.9 (95% CI: 1.26–63.0) times higher in pyretic patients who handled goats than those who handled only cattle, whereas the odds of brucellosis seropositivity were 9.7 (95% CI: 1.28–73.68) times higher in pyretic patients who had backache compared to those without backache. B. abortus DNA was amplified from all six human sera that tested positive by RBT, STAT, and iELISA. As the agreement between the tests was very strong, RBT is recommended as a screening test for the diagnosis of human brucellosis in Bangladesh because it is easier to use, cheaper, and faster. Conclusions: Brucellosis among pyretic patients is common, and B. abortus is responsible for brucellosis in such patients. Pyretic patients who handle goats and those with backaches should be screened for brucellosis.


1998 ◽  
Vol 36 (1) ◽  
pp. 11-14 ◽  
Author(s):  
Paul N. Levett ◽  
Carol U. Whittington

Serology plays an important role in the diagnosis of leptospirosis. Few laboratories have the resources and expertise to perform the microscopic agglutination test. There is a need for rapid and simple serological tests which facilitate the early diagnosis of leptospirosis, while antibiotic therapy may be most effective. A commercially available indirect hemagglutination assay (IHA; MRL Diagnostics, Cypress, Calif.) was evaluated with multiple serum specimens from 107 patients being investigated for leptospirosis. By using a combination of enzyme-linked immunosorbent assay (ELISA) methods for immunoglobulin M (IgM) and IgG antibodies and the microscopic agglutination test, 54 patients were found to have leptospirosis and 53 were found not to have leptospirosis. The sensitivity of IHA for the detection of acute leptospirosis was 100%, the specificity was 94%, the positive predictive value was 95%, and the negative predictive value was 100%. IHA was negative when 13 antinuclear antibody-positive sera, 24 serum specimens from patients with syphilis, and 16 serum specimens false positive by the Venereal Disease Research Laboratory test were tested. IHA was shown to detect both IgM and IgG classes of antibodies in human sera. Serum specimens from 27 dogs investigated for leptospirosis were studied: 3 samples gave nonspecific hemagglutination, but for all remaining samples, the results of IHA and an IgM ELISA were concordant. Performance of IHA was simple, and IHA requires no specialized equipment. It represents a useful assay for laboratories which require a leptospiral diagnostic capability but lack the expertise to perform specialist investigations.


1949 ◽  
Vol 89 (4) ◽  
pp. 369-393 ◽  
Author(s):  
Robert A. Nelson ◽  
Manfred M. Mayer ◽  

Treponema pallida were extracted from rabbit testicular syphilomas and suspended in a special medium in which the organisms remain motile and infectious for several days. On incubation of such suspensions with syphilitic rabbit or human sera and guinea pig complement, the treponemes became non-motile and lost their capacity to infect rabbits. Various factors affecting this immobilization have been investigated. In a preliminary survey of individual sera, immobilizing antibody could be detected in the majority of sera from syphilitic animals and human beings, but was absent in almost all the normal sera examined. It could be demonstrated that the immobilizing and reagin activities of syphilis sera are due to separate antibodies.


1915 ◽  
Vol 21 (6) ◽  
pp. 576-582 ◽  
Author(s):  
Hans Zinsser ◽  
Joseph Gardner Hopkins

It has been shown by our experiments that the serum of rabbits treated with emulsions of Treponema pallidum contains agglutinating substances. Normal rabbit serum also possesses agglutinating power for this organism, but, as in the case of normal bacterial agglutinins, to an extent very much inferior to that possessed by the sera of immunized animals. Normal human sera will agglutinate similar pallidum emulsions, as will the sera of certain syphilitic patients with positive Wassermann reactions. Whether or not there is a quantitative difference of diagnostic value between the sera of normal human beings and those of syphilitics remains to be seen. The sera of rabbits immunized with strain A agglutinate Noguchi's strain 9 in dilutions as high as 1 to 500. We regard as the most important result of these experiments the demonstration of definite antibodies in the circulation of animals treated with dead emulsions of Treponema pallidum. Since it is our belief that the agglutinating effect is due to an antibody essentially the same as that which produces bactericidal, precipitating, and opsonic effects, i. e., that there is probably one type of antibody only, we believe that the demonstration of agglutinins establishes the fact that in syphilis as in bacterial diseases the host responds by the formation of antibodies or sensitizers specific for the treponema. Spirocheticidal experiments with these sera, both in vitro and in vivo, are in progress.


PEDIATRICS ◽  
1949 ◽  
Vol 4 (4) ◽  
pp. 443-453
Author(s):  
ALBERT B. SABIN

An antigen prepared from toxoplasma-infected chorioallantoic membranes, improved by centrifugation at 13,000 r.p.m. for one hour which removed a nonspecific component capable of fixing complement with certain normal sera, proved to be the preparation of choice for toxoplasmic CF tests. Though it was possible to prepare an antigen of similar potency from mouse brain, the yield from one egg is at least 10 times more than from one mouse. The peritoneal exudate of mice freed of Toxoplasma and cells contains eight times as much antigen per unit volume as a 20% extract of mouse brain or chorioallantoic membranes, but its anticomplementary properties, which cannot be removed by high speed centrifugation, can be eliminated only by extensive dilution. The highly centrifuged and diluted peritoneal exudate, however, still contains a component which fixes complement with certain normal human sera. Sixteen children, aged 5 weeks to 11 years, with clinically recognized congenital toxoplasmosis and 17 mothers, with inapparent or clinically unrecognized infection except for having given birth to children with proved toxoplasmosis, were investigated for the presence and persistence of CF antibodies. With the improved antigen and the standardized method that was used, even titers of 1:2 and 1:4 were significant and invariably associated with toxoplasmic neutralizing antibody. The sera of all 17 mothers gave positive CF tests with titers ranging from 1:4 to 1:64, the higher titers predominating during the first two years after the delivery of the infected child. The sera of 14 of the 16 children (87.5%) with clinical evidence of congenital toxoplasmosis, confirmed by neutralization tests, gave positive CF tests with titers ranging from 1:2 to 1:128, the higher titers again predominating during the first two years after birth. One of the negative results, in a 5 week old child with active infection who died three days after the serum was obtained, was associated with a high (1:4,096) dye test titer for neutralizing antibody in its own serum, and a similar dye test titer (1:4,096) and a CF titer of 1:32 in the serum of its mother. The other negative result, in a 7 year old child with a dye test titer of 1:16, is believed to represent an instance of disappearance of CF antibody after an interval of seven years. Not one serum obtained from 24 children with congenital ocular or neurologic disturbances or both, not due to toxoplasmosis, gave a positive CF test with the toxoplasmic antigen; however, among 20 of their mothers, there were four sera with titers of 1:2, 1:2, 1:8 and 1:16. Toxoplasmic CF antibody can persist for at least six years, and in some instances even longer, in individuals with clinically recognized as well as inapparent or clinically unrecognized infections. Since at least seven of the mothers, who gave birth to children with congenital toxoplasmosis, subsequently gave birth to normal children at a time when the CF antibody titers in some of them were still high (32, 32, 32, 16, 8, 4, 4), it is clear that this antibody can persist in individuals in whom the infection has been eradicated or suppressed sufficiently to prevent its congenital transmission.


Author(s):  
Cong Ding ◽  
Yufei Zhang ◽  
Chen Chuan ◽  
Yanhua Liu

Indoor air pollution seriously threats the life and health of human beings. The improvement of indoor air quality has become a focus that people pay more and more attentions to. The photocatalytic of pollutants based on TiO2 is a promising air purification technology. In order to overcome the disadvantages of nanometer powder TiO2 catalyst and to enhance the photocatalytic activity of TiO2, series of glass plates covered with doped-TiO2 were prepared and the photocatalysis them were studied. The glass plates covered with TiO2 which was doped in advance with N, F, or/and Fe were prepared by a sol-gel method. The doping content of N, F, Fe and heat treatment temperature were determined using the orthogonal array of the Taguchi quality design. The prepared gel was coated on the glass by spin-coating method. The effects of doping level of N, F and Fe and heat treatment temperature on the photocatalytic capabilities were investigated. The photocatalytic capabilities of prepared glass plates were investigated by degrading the solution of methylene blue (MB,C16H18ClN3S). The results show that appropriate addition of N, F and Fe and temperaturae are effective for improving the photocatalytic activities of TiO2 under visible light. The optimal TiO2 was prepared under the condition that the doping amount of F element was 9at %, that of N is 7at %, and none of Fe under 400 °C calcination temperature. The degradation rate of the sample for methylene blue solution reaches 23.49% under visible light irradiation for 5 hours. The influence order of the factors was the calcination temperature > F > N > Fe.


2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Xiao Teng Ching ◽  
Yee Ling Lau ◽  
Mun Yik Fong ◽  
Veeranoot Nissapatorn ◽  
Hemah Andiappan

Toxoplasma gondiiinfects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of wholeToxoplasmalysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressedT. gondiidense granular protein-5 (GRA5) inEscherichia coliand isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronicT. gondiiinfections (sensitivities of 46.8% and 61.2%, resp.).


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