The cloning, expression, and nucleotide sequence of a gene coding for an immunogenic Brucella abortus protein

Mayfield, John E.; Bricker, Betsy J.; Godfrey, Henry P.; Crosby, Renae M.; Knight, D. J. E.; Halling, Shirley M.; Balinsky, Doris; Tabatabai, Louisa B.

doi:10.1016/0378-1119(88)90540-9

Cited by 101 publications

(82 citation statements)

References 20 publications

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“…This membrane protein is known as a Brucella gender (Mayfield et al, 1988). Furthermore, our isolates have been amplified by specific primers for B. abortus, B. melitensis, B. ovis, B. suis biovar 1 and B. suis biovar 2 (in a reference laboratory).…”

Section: Discussionmentioning

confidence: 99%

Wild boars (Sus scrofa) as reservoirs of Brucella suis biovar 2 in Croatia

Cvetnić

Mitak

Ocepek

et al. 2003

Acta Veterinaria Hungarica

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This work presents the results of findings for brucellosis in wild boars and domestic swine in two regions of Croatia. In the region of Djakovo the blood samples of 211 wild boars were analysed and in 29.4% of the samples serologically positive reactions were established. In the same region the blood samples of 1080 domestic swine on pastures were also analysed and positive serological reactions were established in 12.3%. In the regions around Lonjsko Polje the blood samples of 53 wild boars were analysed and in 22.6% of them positive serological reactions were established. On several locations around Lonjsko Polje the blood samples of 901 domestic swine were serologically analysed and 13.5% of the swine were found to be seropositive. Bacteriological analyses of submitted materials from 24 wild boars resulted in isolation of Brucella from seven (29.2%) samples, and from 43 samples originating from domestic swine that had aborted and had been serologically positive, Brucella were isolated from 25 (58.1%) swine, as well as from 10 (62.5%) out of 16 aborted piglets. In all the isolates Brucella suis biovar 2 was identified. Wild boars are carriers and reservoirs of Brucella suis biovar 2 in Croatia.

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Section: Discussionmentioning

confidence: 99%

Wild boars (Sus scrofa) as reservoirs of Brucella suis biovar 2 in Croatia

Cvetnić

Mitak

Ocepek

et al. 2003

Acta Veterinaria Hungarica

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“…The BCSP 31 is a membrane antigen characteristic for Brucella genus (Mayfield, 1988). The primers we used were BRU-UP (GGG CAA GGA AGA TTT) and BRU-LOW (CGG CAA GGG TCG GTG TTT) (Qiagen Operon, Germany) that allowed the amplification of a fragment of approximately 443 bp (Serpe et al, 1999).…”

Section: Isolates Identificationmentioning

confidence: 99%

Brucellosis in wild boar (Sus scrofa) in the Republic of Croati

Cvetnić¹,

Tončić²,

Špičić³

et al. 2004

Vet. Med.

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During the years 2001 and 2002 on seven localities in Croatia a survey on the prevalence of brucellosis in wild boar was carried out. The survey included 271 (52.7%) female and 243 (47.3%) male animals between 7 months and 4 years of age and weighing from 14 to 135 kg. On that occasion 514 blood samples of wild boar were serologically analysed. For serological analysis indirect enzyme immunoassay (iELISA), Rose Bengal test (RBT), complement fixation test (CFT) and slow agglutination test (SAT) were used. In all of the wild boar from all of the localities investigated positive reactions to brucellosis were established. Most of the positive reactions were established by iELISA (13.6%), then by RBT (11.5%), CFT (10.5%) and SAT (8.9%). Tissue samples of 106 animals: testes samples from 67 animals, uterus tissue from 38 animals and 5 fetuses of piglets from 1 mother were analysed bacteriologically. Brucella suis biovar 2 was isolated from 18 (17.0%) animals that originated from all of the localities investigated. Isolates were identified by PCR using BRU-UP and BRU-LOW primers specific for genus Brucella and primers specific for IS711. Based on our results it could be concluded that in Croatia wild boar are natural vector and/or reservoirs of B. suis biovar 2. This permanent risk factor is hazardious for domestic and wild animals in the Republic of Croatia.

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“…Of the 17 blood samples obtained from patients with brucellosis, eight were positive by culture (Table 1), with a mean detection time of 7 days (range [4][5][6][7][8][9][10][11][12][13][14]. All the samples showed high-titer antibrucella antibodies, including the three samples submitted after treatment.…”

Section: Resultsmentioning

confidence: 99%

“…8 PCR was performed using a protocol described elsewhere. 4 Briefly, a 50 μL volume reaction mixture containing 10 mM tris-HCl (pH 8.4), 50 mM KCl, 1 mM MgCl 2 , 200 μM each deoxyribonucleotide triphosphate (dATP, dGTP, dTTP, dCTP, Pharmacia LKB Biotechnology, Uppsala, Sweden), 1.5 U of Taq polymerase (Perkin-Elmer Cetus Co., Norwalk Conn., USA), oligonucleotide B4 and B5 (100 nM each; Gulf Biotech, Riyadh, Saudi Arabia) and 2-4 μg of total DNA extracted or 150 ng from the positive control was processed in a thermocycler (Perkin-Elmer 9600).…”

Section: Dna Amplificationmentioning

confidence: 99%

Evaluation of PCR, Culture and Serology for the Diagnosis of Acute Human Brucellosis

et al. 2000

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Background:The diagnosis of brucellosis is frequently difficult to establish. This is not only because clinically, the disease can mimic any infectious and noninfectious disease, but also because the established diagnostic methods are not always successful. In this study, we have tried to evaluate PCR techniques in the diagnosis of brucellosis in comparison to conventional techniques. Patients and Methods: Fifty peripheral blood samples from the following groups were collected: patients with brucellosis (17); patients with febrile illnesses due to factors other than brucella etiology (19); symptomatic occupationally exposed persons (9); and healthy volunteers (5). The last three groups were considered controls. Among the 17 Brucella samples, only 14 were obtained before treatment was begun. The samples were tested by serology, using the standard tube agglutination method (STA), blood culture using Bactec machines, and PCR using primer pair to amplify a 223-bp region within a gene coding for a 31-kD Brucella antigen. Diagnosis of brucellosis was based on compatible clinical picture in addition to positive blood culture and/or positive serology. Results: Of the 17 blood samples from patients with brucellosis, eight were culture positive for Brucella species, and all showed high titer antibrucella antibodies. Only 14 of them were positive by PCR, and these were the samples submitted before initiation of therapy, representing 100% sensitivity. Among the 33 controls, blood culture was negative for Brucella in all of them, while one sample showed high-titer antibrucella antibodies. The latter was from the febrile illnesses group. PCR-based assay was able to detect four bands in the controls, all of which were from the occupationally exposed asymptomatic group. Conclusion: In view of the several advantages of PCR over the conventional methods for the diagnosis of brucellosis, such as speed, safety, high sensitivity and specificity, the technique might be considered for laboratory diagnosis of brucellosis. However, for the evaluation of asymptomatic highly exposed persons, PCR might be considered complementary to the traditional methods and followed up by serology and/or culture.

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The cloning, expression, and nucleotide sequence of a gene coding for an immunogenic Brucella abortus protein

Cited by 101 publications

References 20 publications

Wild boars (Sus scrofa) as reservoirs of Brucella suis biovar 2 in Croatia

Wild boars (Sus scrofa) as reservoirs of Brucella suis biovar 2 in Croatia

Brucellosis in wild boar (Sus scrofa) in the Republic of Croati

Evaluation of PCR, Culture and Serology for the Diagnosis of Acute Human Brucellosis

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