Diagnosis of canine brucellosis by detection of serum antibodies against an 18 kDa cytoplasmic protein of Brucella spp.

Baldi, Pablo C.; Wanke, M.M.; Loza, Maria E.; Monachesi, Norma Estela; Fossati, Carlos A.

doi:10.1016/s0378-1135(97)00134-x

Cited by 31 publications

(32 citation statements)

References 9 publications

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“…Interestingly, the determination of the antibody response to this protein yields results equivalent to those obtained with a complex mixture of cytoplasmic proteins of Brucella (5,7,8).…”

Section: Discussionmentioning

confidence: 84%

“…Antibodies to the 18-kDa protein were found in 26 animals which were also positive for anti-CP antibodies and for anti-LPS antibodies (the latter detected by the slide agglutination test). The overall correlation between the two anti-protein ELISAs reached 93.3% (8).…”

Section: Diagnostic Usefulness Of Brucella Cytoplasmic Proteinsmentioning

confidence: 90%

“…While the latter had high levels of antibodies against the 18-kDa protein, the vaccinated cattle developed only a low and transient antibody response that was no longer detected at 90 days post-vaccination (a time when anti-LPS antibodies were still detected at high levels). The 18-kDa protein and the CP antigen were also useful for the diagnosis of canine brucellosis (6,8). The antibody reactivity to cytoplasmic proteins of Brucella was investigated by ELISA in sera from 30 dogs having confirmed or suspected brucellosis.…”

Section: Diagnostic Usefulness Of Brucella Cytoplasmic Proteinsmentioning

confidence: 99%

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Structural, functional and immunological studies on a polymeric bacterial protein

Baldi

Velikovsky

Braden

et al. 2000

Braz J Med Biol Res

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The characterization of proteins from Brucella spp, the causative agent of brucellosis, has been the subject of intensive research. We have described an 18-kDa cytoplasmic protein of Brucella abortus and shown the potential usefulness of this protein as an antigen for the serologic diagnosis of brucellosis. The amino acid sequence of the protein showed a low but significant homology with that of lumazine synthases. Lumazine is an intermediate product in bacterial riboflavin biosynthesis. The recombinant form of the 18-kDa protein (expressed in E. coli) folds like the native Brucella protein and has lumazinesynthase enzymatic activity. Three-dimensional analysis by X-ray crystallography of the homolog Bacillus subtilis lumazine synthase has revealed that the enzyme forms an icosahedral capsid. Recombinant lumazine synthase from B. abortus was crystallized, diffracted X rays to 2.7-Å resolution at room temperature, and the structure successfully solved by molecular replacement procedures. The macromolecular assembly of the enzyme differs from that of the enzyme from B. subtilis. The Brucella enzyme remains pentameric (90 kDa) in its crystallographic form. Nonetheless, the active sites of the two enzymes are virtually identical at the structural level, indicating that inhibitors of these enzymes could be viable pharmaceuticals across a broad species range. We describe the structural reasons for the differences in their quaternary arrangement and also discuss the potential use of this protein as a target for the development of acellular vaccines. Correspondence

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“…Interestingly, the determination of the antibody response to this protein yields results equivalent to those obtained with a complex mixture of cytoplasmic proteins of Brucella (5,7,8).…”

Section: Discussionmentioning

confidence: 84%

Section: Diagnostic Usefulness Of Brucella Cytoplasmic Proteinsmentioning

confidence: 90%

Section: Diagnostic Usefulness Of Brucella Cytoplasmic Proteinsmentioning

confidence: 99%

See 1 more Smart Citation

Structural, functional and immunological studies on a polymeric bacterial protein

Baldi

Velikovsky

Braden

et al. 2000

Braz J Med Biol Res

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“…Agar gel immunodiffusion (AGID) has been used but sometimes the precipitin lines are difficult to interpret. More recently, indirect enzymelinked immunoassay (IELISA) with cytoplasmic protein antigen, hot saline extracts or cell-wall antigens have been proposed [2][3][4][5].…”

Section: Introductionmentioning

confidence: 99%

Sensitivity and specificity of an indirect enzyme-linked immunoassay for the diagnosis of Brucella canis infection in dogs

Lucero¹,

Escobar²,

Ayala³

et al. 2002

Journal of Medical Microbiology

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The diagnosis of B. canis infection in dogs is based on bacteriological examination and serological methods including agglutination and gel diffusion tests. Bacteriological studies are the only methods that have been considered specific but, as intermittent periods of abacteraemia may occur, a negative blood culture cannot be used as a criterion for excluding canine brucellosis. Close contact between people and infected dogs increases the risk of transmission; however, its impact on public health is probably underestimated due to lack of reporting and inadequate diagnostic services. This paper describes an indirect enzyme-linked immunoassay (IELISA) procedure for the diagnosis of brucellosis caused by B. canis in a population of normal and infected dogs previously screened by the buffered plate antigen test (BPAT) and rapid slide agglutination test (RSAT). The serological survey was performed with 446 field sera. The 270 sera from the asymptomatic group found negative by BPA, RSAT and blood culture showed IELISA specificities of 96.7% and 100%, respectively, when cut-off values of OD 0.237 and 0.281 were selected. For 52 sera from culture-positive dogs, IELISA sensitivity was 100% with cut-off values of OD 414 0.237 and 0.281. OD 414 0.281 was selected because this value provided the highest accuracy with minimal false-negative and false-positive results. This cut-off value was used to study 124 blood culture-negative but RSAT positive sera. IELISA produced 107 positive results; the 17 sera that were negative by IELISA presented a wide range of reactivities by RSAT (2 were RSAT positive at 1 in 2 dilution and 15 were weakly positive with pure serum). These samples were probably from animals at an early stage of the infection or were false-positive results. The IELISA described here detects IgG and IgA antibodies that are useful for evaluating the clinical status of dogs. Although RSAT is a practical screening test, a supplementary technique such as IELISA should be used on all positive RSAT samples to ensure diagnostic specificity. Furthermore, people in contact with infected dogs could be investigated for possible transmission. The procedure described in this study was relatively simple and could have widespread applications.

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“…Uma desvantagem das provas baseadas em antígenos internos é a detecção de anticorpos produzidos contra brucelas lisas e rugosas, não possibilitando a identificação da espécie de Brucella responsável pela infecção (CARMICHAEL, 1990;JONES, 1989;SHIN, 1996; (BALDI et al, 1994(BALDI et al, , 1997WANKE;DELPINO;BALDI, 2002).…”

Section: Transmissãounclassified

Avaliação de métodos diretos e indiretos de diagnóstico da brucelose em cães naturalmente infectados

Keid¹

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.Foram comparados procedimentos laboratoriais diretos e indiretos aplicados ao diagnóstico da brucelose canina causada por Brucella canis. Foram examinados 196 cães, os quais foram classificados em infectados, suspeitos e não infectados, de acordo com resultados obtidos no cultivo microbiológico em amostras de sangue, sêmen e swab vaginal, no exame clínico e de acordo com dados epidemiológicos. Os resultados obtidos nos exames laboratoriais foram correlacionados com os resultados dos exames clínicos. As técnicas laboratoriais empregadas foram soroaglutinação rápida (SAR), soroaglutinação rápida com emprego do 2-mercaptoetanol (SAR-2ME), imunodifusão em gel de ágar (IDGA), cultivo microbiológico e PCR em amostras de sangue, sêmen e swab vaginal. Foram observados 41,83% de positivos pela SAR, 14,28% pela SAR-2ME, 28,14% pela IDGA, 32,14% pela hemocultura, 19,6% pelo cultivo microbiológico de sêmen, 4,82% pelo cultivo microbiológico de swab vaginal, 33,16% pela PCR em sangue, 33,33% pela PCR em sêmen e 26,89% pela PCR em swab vaginal. Os valores de sensibilidade das provas de SAR, SAR-2ME, IDGA, PCR em sangue e PCR em swab vaginal foram respectivamente de 81,25%; 42,18%; 75%; 89,06% e 54%. A especificidade dos métodos de SAR, SAR-2ME, IDGA, PCR em sangue e PCR em swab vaginal foram respectivamente de 81,25%; 100%; 100%; 97,21% e 97,14%. Nos cães machos, as proporções de resultados positivos diagnosticados pelo cultivo microbiológico de sêmen foram semelhantes às observadas pelos demais métodos de diagnóstico utilizados. As proporções de resultados positivos obtidos pelas técnicas de SAR, SAR-2ME, IDGA, hemocultura, PCR em sangue e cultivo de swab vaginal foram maiores dentre os animais que apresentaram sinais clínicos sugestivos de brucelose. Não houve associação entre a presença de sinais clínicos de brucelose e resultados positivos pelo cultivo microbiológico de sêmen e PCR em amostras de sêmen e swab vaginal.

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Diagnosis of canine brucellosis by detection of serum antibodies against an 18 kDa cytoplasmic protein of Brucella spp.

Cited by 31 publications

References 9 publications

Structural, functional and immunological studies on a polymeric bacterial protein

Structural, functional and immunological studies on a polymeric bacterial protein

Sensitivity and specificity of an indirect enzyme-linked immunoassay for the diagnosis of Brucella canis infection in dogs

Avaliação de métodos diretos e indiretos de diagnóstico da brucelose em cães naturalmente infectados

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