Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308

Matrone, M.; Keid, Lara Borges; Rocha, V.C.M.; Vejarano, Maria P; Ikuta, Cássia Yumi; Rodriguez, C.A.R.; Ferreira, Fernando; Dias, Ricardo Augusto; Neto, José Soares Ferreira

doi:10.1590/s1517-83822009000300010

Cited by 20 publications

(8 citation statements)

References 14 publications

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“…Thus, high concentrations of Tris, EDTA, and SDS, with increased incubation temperature, were used in our study. The present study included a phenol-chloroform step at the end of the extraction protocol and it had a protein impurity reducing effect on total genomic DNA, which conflicts with some of previous reports (25,26) and confirms the findings of Matrone et al (27). Therefore, these data indicate that the modified DNA extraction protocol used in our study can also be applied for PCR detection of many other bacterial pathogens in milk.…”

Section: Discussionsupporting

confidence: 53%

PCR detection of Brucella abortus in cow milk samples collected from Erzurum, Turkey

Arasoğlu¹,

Güllüce²,

Özkan³

et al. 2013

Turk J Med Sci

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Exposures of Upper Jurassic reef outcrops in the Russian western Caucasus provide excellent fi eld analogues for possible reef-complex reservoir targets imaged on seismic refl ection data from the northern Shatskiy Ridge, eastern Black Sea. Th e reefs at outcrop can be generally grouped into coral-dominated, siliceous sponge-microbialite and microbialite types. Coral-dominated reefs occur as isolated patchy and massive forms, and can be subdivided into higher-diversity and low-diversity types. Th e former developed at shallow-water platform margins and in platform interiors, whilst the latter occurred in deeper-water mid-shelf settings. Siliceous sponge-microbialite and microbialite reefs occur as lenses and mounds that were restricted to deeper-water mid-outer shelf environments. Th e reefs developed on two Late Jurassic carbonate platforms in the Russian western Caucasus: the north Caucasus platform to the north and South Adler platform to the south. Th ese platforms were separated by a deep marine (Greater Caucasus) basin, along the margins of which thick shallow-water coral-dominated reefs formed. Th e southwestern margin of the north Caucasus platform probably represented a reef barrier-slope-basinal system that was structurally controlled. At the northeastern margin of this platform, in the Laba River region, inner ramp coral-dominated reefs pass northwards into deeper-water siliceous sponge-and microbialite-dominated reefs. Th e South Adler platform may extend off shore into the eastern Black Sea and the coral-dominated reefs that crop out at its northern margin form potential exploration analogues. Th e palaeowater depth of the possible isolated reef complexes imaged on the Shatskiy Ridge is unclear. If they were deposited in shallow-water, the coral-dominated reefs examined in the north Caucasus or South Adler carbonate platforms may form suitable analogues. Alternatively, if they were developed in deeper water they may be equivalent to the siliceous sponge and microbialite reefs examined in the Laba River region.

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Section: Discussionsupporting

confidence: 53%

PCR detection of Brucella abortus in cow milk samples collected from Erzurum, Turkey

Arasoğlu¹,

Güllüce²,

Özkan³

et al. 2013

Turk J Med Sci

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“…DNA extraction from semen and urine was performed using the proteinase K and phenol/ chloroform method, with 500 μL and 1 mL, of semen and urine, respectively [31]. DNA extraction from blood and vaginal swabs was performed using the guanidine method [32], with 250 μL of each sample.…”

Section: Dna Extraction and Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Brucella ovis mutant in ABC transporter protects against Brucella canis infection in mice and it is safe for dogs

et al. 2020

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Background/Objectives Vaccination is the most important tool for controlling brucellosis, but currently there is no vaccine available for canine brucellosis, which is a zoonotic disease of worldwide distribution caused by Brucella canis. This study aimed to evaluate protection and immune response induced by Brucella ovis ΔabcBA (BoΔabcBA) encapsulated with alginate against the challenge with Brucella canis in mice and to assess the safety of this strain for dogs. Methods Intracellular growth of the vaccine strain BoΔabcBA was assessed in canine and ovine macrophages. Protection induced by BoΔabcBA against virulent Brucella canis was evaluated in the mouse model. Safety of the vaccine strain BoΔabcBA was assessed in experimentally inoculated dogs. Results Wild type B. ovis and B. canis had similar internalization and intracellular multiplication profiles in both canine and ovine macrophages. The BoΔabcBA strain had an attenuated phenotype in both canine and ovine macrophages. Immunization of BALB/c mice with alginateencapsulated BoΔabcBA (10 8 CFU) induced lymphocyte proliferation, production of IL-10 and IFN-γ, and protected against experimental challenge with B. canis. Dogs immunized with alginate-encapsulated BoΔabcBA (10 9 CFU) seroconverted, and had no hematologic, biochemical or clinical changes. Furthermore, BoΔabcBA was not detected by isolation or PCR performed using blood, semen, urine samples or vaginal swabs at any time point over the course of this study. BoΔabcBA was isolated from lymph nodes near to the site of inoculation in two dogs at 22 weeks post immunization.

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“…DNA extraction was performed by the proteinase K and phenol/chlorophorm method as previously described [18] with 500 μL of fresh semen or blood samples, 1 mL of thawed urine or preputial wash samples, and approximately 800 μL of tissue homogenates. All DNA samples were stored at −20°C until amplification.…”

Section: Methodsmentioning

confidence: 99%

Species-specific multiplex PCR for the diagnosis of Brucella ovis, Actinobacillus seminis, and Histophilus somni infection in rams

et al. 2013

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BackgroundInfectious ovine epididymitis results in substantial economic losses worldwide due to reproductive failure and culling of breeders. The most common causative agents of these infections are Brucella ovis, Actinobacillus seminis, and Histophilus somni. The aim of this study was to develop a multiplex PCR assay for simultaneous detection of Brucella ovis, Actinobacillus seminis, and Histophilus somni with species-specific primers applied to biological samples for molecular diagnosis of these infections.ResultsThe multiplex assay was capable of detecting B. ovis, A. seminis, and H. somni DNA simultaneously from genomic bacterial DNA samples and pool of semen samples from experimentally infected rams. The method was highly specific since it did not amplify DNA from other bacterial species that can potentially cause epididymitis in rams as well as species phylogenetically related to B. ovis. All negative control samples were negative in PCR multiplex assay. Urine can be used as an alternative to semen samples.ConclusionsThe species-specific multiplex PCR assay developed in this study can be successfully used for the detection of three of the most common bacterial causes of ovine epididymitis.

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Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308

Cited by 20 publications

References 14 publications

PCR detection of Brucella abortus in cow milk samples collected from Erzurum, Turkey

PCR detection of Brucella abortus in cow milk samples collected from Erzurum, Turkey

Brucella ovis mutant in ABC transporter protects against Brucella canis infection in mice and it is safe for dogs

Species-specific multiplex PCR for the diagnosis of Brucella ovis, Actinobacillus seminis, and Histophilus somni infection in rams

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