Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams.

Brucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. The goal of this study was to develop a PCR test targeting specific B. ovis genomic sequences. Specific primer pairs were designed targeting 12 of those ORFs. Samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams (n=9) every other week up to 180 days post infection (dpi), when tissue samples were obtained. Blood, serum, semen, urine, and preputial wash samples were obtained, in weekly intervals for 1 month, from eight rams belonging to a B. ovis-free flock. Semen samples were also obtained from rams belonging to naturally infected flocks (n=40). The limit of detection of this PCR protocol was 100, 10, and 1 CFU/mL for semen, urine and prepucial wash samples, respectively. Sensitivity and specificity values obtained with this PCR method were similar to that of bacteriology when evaluating biological samples. Agreement between PCR and bacteriology results was greater than 90%. These results clearly indicate that this species-specific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.

Rapid diagnosis of Brucella epididymo-orchitis by real-time polymerase chain reaction assay in urine samples.

PURPOSE: We studied the diagnostic yield of a real-time polymerase chain reaction assay in urine samples for the rapid diagnosis of brucella epididymo-orchitis compared to that of conventional microbiological techniques. MATERIALS AND METHODS: We used an SYBR Green I LightCycler based real-time polymerase chain reaction to retrospectively study 10 urine samples from patients with Brucella epididymo-orchitis. The assay amplifies a 223 bp sequence of a gene that codes for the synthesis of an immunogenetic membrane protein specific for Brucella genus (BCSP31). After amplifying this 223 bp sequence we performed melting curve analysis to verify the specificity of polymerase chain reaction products. RESULTS: Brucella melitensis was isolated from blood cultures in 9 cases (90%). Wright's seroagglutination was negative or inconclusive in 30% of cases. Brucella was isolated from urine in only 1 case, whereas real-time polymerase chain reaction assay in urine was positive in 9 (90%). Also, results were available in 4 hours, whereas mean time to availability of the final blood culture results was 5.8 days (range 4.5 to 7). CONCLUSIONS: SYBR Green I LightCycler based real-time polymerase chain reaction assay in urine samples is highly sensitive and specific, and easy to perform. It could provide the clinician with results in less than 5 hours. The technique could be a practical and useful tool for the rapid diagnosis of genitourinary complications of human brucellosis.

Perfil sorológico, isolamento bacteriano e valores hematológicos e urinários em cäes naturalmente infectados com Brucella canis / Serology, bacterial isolation, hematological and urinary values in dogs naturally infected with Brucella canis

Descrevem-se os parâmetros hematológicos, urinários, perfil sorológico de aglutininas antibrucélicas e resultados de isolamento bacteriano de swab vaginal, líquido prostático e hemocultura de 12 cäes naturalmente infectados por Brucella canis. Observaram-se flutuaçäo dos resultados sorológicos, ausência de isolamento de B. canis nos diversos materiais colhidos e valores hematológicos e urinários predominantemente normais. Discute-se o diagnóstico de brucelose canina em nível individual.

Acute brucellosis associated with massive proteinuria.

Mild proteinuria during these renal complications in brucellosis is a common feature, however, heavy proteinuria in association with renal brucellosis has rarely been reported and it always was associated with a concomitant development of renal failure. This is the first description of heavy proteinuria as the sole renal derangement during acute infection with brucellosis.

Brucellosis in man--II. Isolation of the causative organisms with special reference to blood picture and urine constituents.

Blood and urine samples were collected from 50 patients with febrile splenomegaly suspected to be Malta fever in Sharkia fever hospitals, Egypt. Blood samples were subjected to isolation of Brucellae; blood counts and erythrocyte sedimentation rate. Urine analysis. Four strains of brucella were isolated. Two of these isolated first from the 50 patients and typed as Br. melitensis Biotype I and the others were recovered secondly and thirdly from serologically positive patients and typed as Br. abortus Biotype I. All the strains from positive serology, titre ranging from 1/160-1/640. Br. melitensis is more readily isolated from the blood than Br. abortus. Blood picture of serologically positive Malta fever showed three patients with mild leukopenia; two with relative lymphocytosis; two with secondary anaemia and three with accelerated erythrocyte sedimentation rate. Urine analysis revealed five patients with albuminuria.