Potency of partially purified malleo-proteins for mallein test in the diagnosis of glanders in equines
TL;DR: Ultrogel AcA 34 column fractionation studies revealed that mallein activity was associated with high molecular weight proteins and represented a purer sensitin than PPD mallein for the purpose of mallein test.
Abstract: Malleo-proteins from synthetic broth mallein of six strains of Pseudomonas mallei were separated by trichloroacetic acid precipitation, amonium sulfate precipitation and Ultrogel AcA 34 gel filtration chromatography. When tested comparatively with Dutch PPD mallein as standard on P. mallei -sensitized and normal horses all the strains were found to be malleinogenic, trichloroactive acid precipitated proteins were comparable to Dutch PPD mallein in potency and innocuity whereas ammonium sulfate-precipitated proteins elicited non-specific reactions. Ultrogel AcA 34 chromatographed high molecular weight proteins (MW range > 350 000) were having equal or higher potency and without adverse reactions and low molecular weight proteins (MW range 120 000–170 000) nearly inactive. Ultrogel AcA 34 column fractionation studeis revealed that mallein activity was associated with high molecular weight proteins and represented a purer sensitin than PPD mallein for the purpose of mallein test.
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TL;DR: A literature review of human glanders is presented in which it is discussed the clinical epidemiology and risk factors, potential routes of exposure, symptoms, the incubation period, and specific diagnostics, and a clinical definition of human pulmonary glanders infection is proposed.
Abstract: Glanders is a highly contagious and often fatal zoonotic disease, primarily of solipds. In the developed world, glanders has been eradicated. However, prior use of B. mallei as a biological weapon and its high mortality in inhalation animal studies has affirmed B. mallei as a biodefense concern. This threat requires the development of new glanders medical countermeasures (MCMs), as there is a lack of an effective vaccine and lengthy courses of multiple antibiotics needed to eradicate B. mallei. Here, we present a literature review of human glanders in which we discuss the clinical epidemiology and risk factors, potential routes of exposure, symptoms, the incubation period, and specific diagnostics. This review focuses on pulmonary glanders, as this is the most likely outcome of a biological weapons attack. Additionally, we outline current treatment regimens and propose a clinical definition of human pulmonary glanders infection.
93 citations
Cites methods from "Potency of partially purified malle..."
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TL;DR: The Rose Bengal plate agglutination test was evaluated for the diagnosis of equine glanders, and its diagnostic efficiency was compared with that of mallein and other serological tests, including indirect hemagglutination Test (IHAT), complement fixation test (CFT), and modified counter immunoelectrophoresis test (mCIET).
Abstract: The Rose Bengal plate agglutination test (RBT) was evaluated for the diagnosis of equine glanders, and its diagnostic efficiency was compared with that of mallein and other serological tests, including indirect hemagglutination test (IHAT), complement fixation test (CFT), and modified counter immunoelectrophoresis test (mCIET). Sera from 70 naturally infected culture-positive, 96 potentially exposed cohorts, and 110 healthy equines were tested. All tests but mCIET showed 100% specificity when testing the sera from glanders-negative equines. The calculated sensitivities of RBT, IHAT, CFT, mCIET, and mallein test when testing culture-positive equines were 90.0, 97.1, 91.4, 81.4, and 75.7%, respectively. The RBT was significantly (P < 0.05) more sensitive than the mallein test and mCIET. The positive and negative predictive values of each test (RBT, IHAT, CFT, mallein test, and mCIET) were as follows: 100 and 94, 100 and 98.2, 100 and 96.7, 100 and 86.6, and 90.5 and 88.6, respectively. On comparing glandered and nonglandered animals, the highest agreement (0.987) was found between RBT and CFT followed by RBT and IHAT (0.940), RBT and mallein test (0.871), and RBT and mCIET (0.852). Because the RBT is simpler and rapid to perform, the inclusion of the test as a supplementary test for the diagnosis of glanders in field conditions is recommended.
52 citations
Cites background from "Potency of partially purified malle..."
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TL;DR: The results of this study showed that glanders in the region was emerging and further sero-epidemiological studies are urgent for the eradication of the disease.
Abstract: Based on clinical-pathological, epidemiological and serological aspects, as well as by the isolation of Burkholderia mallei, foci of glanders were diagnosed in equids of the "Zona da Mata" in the states of Pernambuco and Alagoas. Clinically there was hyperthermia, noisy respiration, loss of appetite, progressive loss of weight leading to emaciation, nasal congestion and muco-purulent discharge, erosions, ulcers and scars of the nasal mucosa. The size of submaxillary and cervical lymphonodes was increased, which contained abscesses and fistules exudating purulent material. In the skin, accompanying the lymph vessels, there were firm nodules and abscesses which in chronic cases fistulated and ulcerated leaving stellar scars. The histopathology of the nodules at various sites, from the horses as well as from the guinea-pigs used for the Strauss test, showed granulomatous lesions and pyogranulomas. The results of this study showed that glanders in the region was emerging and further sero-epidemiological studies are urgent for the eradication of the disease.
34 citations
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01 Jan 2000
TL;DR: Mota et al. as mentioned in this paper showed that glanders in the region was emerging and further sero-epidemiologicalstudies are urgent for the eradication of the disease.
Abstract: .- Mota R.A., Brito M.F., Castro F. J.C. & Massa M. 2000. [Glanders in horses andmules of the states of Pernambuco and Alagoas, Brazil.]ernambuco and Alagoas, Brazil. Mormo em equideos nos Estadosde Pernambuco e Alagoas. Pesquisa Veterinaria Brasileira 20(4):155-159. Depto Med. Vet., Univ.Fed. Rural de Pernambuco, Rua Dom Manoel de Medeiros s/no, Dois Irmaos, Recife, PE 52171-900, Brazil.Based on clinical-pathological, epidemiological and serological aspects, as well as by theisolation of Burkholderia mallei, foci of glanders were diagnosed in equids of the “Zona daMata” in the states of Pernambuco and Alagoas. Clinically there was hyperthermia, noisyrespiration, loss of appetite, progressive loss of weight leading to emaciation, nasal congestionand muco-purulent discharge, erosions, ulcers and scars of the nasal mucosa. The size ofsubmaxillary and cervical lymphonodes was increased, which contained abscesses and fistulesexudating purulent material. In the skin, accompanying the lymph vessels, there were firmnodules and abscesses which in chronic cases fistulated and ulcerated leaving stellar scars.The histopathology of the nodules at various sites, from the horses as well as from the guinea-pigs used for the Strauss test, showed granulomatous lesions and pyogranulomas. The resultsof this study showed that glanders in the region was emerging and further sero-epidemiologicalstudies are urgent for the eradication of the disease.
31 citations
References
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TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
Abstract: Since 1922 when Wu proposed the use of the Folin phenol reagent for the measurement of proteins, a number of modified analytical procedures utilizing this reagent have been reported for the determination of proteins in serum, in antigen-antibody precipitates, and in insulin. Although the reagent would seem to be recommended by its great sensitivity and the simplicity of procedure possible with its use, it has not found great favor for general biochemical purposes. In the belief that this reagent, nevertheless, has considerable merit for certain application, but that its peculiarities and limitations need to be understood for its fullest exploitation, it has been studied with regard to effects of variations in pH, time of reaction, and concentration of reactants, permissible levels of reagents commonly used in handling proteins, and interfering substances. Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
285,427 citations
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TL;DR: Dot ELISA had the highest sensitivity, and was superior to other tests in that it was rapid and easy to perform, the results were easy to interpret, the assay was not influenced by anti-complement activity, and it was able to detect antibodies at an early stage.
Abstract: A dot enzyme-linked immunosorbent assay (dot ELISA) was developed for diagnosis of glanders in equines. The test was based on the detection of IgG antibodies to Pseudomonas mallei antigens bound to nitrocellulose coated on plastic strips (dipsticks), the reaction being amplified by an avidinbiotin system with biotinylated anti-horse IgG and horseradish peroxidase-avidin D. Sera from 810 normal, six naturally infected and 48 sensitized equines were tested by this assay, and results were compared with complement fixation, indirect haemagglutination and counter-immunoelectrophoresis tests. Dot ELISA had the highest sensitivity, and was superior to other tests in that it was rapid and easy to perform, the results were easy to interpret, the assay was not influenced by anti-completement activity, and it was able to detect antibodies at an early stage. Testing of serum at 1:200 dilution is proposed for epidemiological screening.
33 citations
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