Alternative PCR protocol using a single primer set for assessing DNA quality in several tissues from a large variety of mammalian species living in areas endemic for leishmaniasis

Ferreira, E.; Gontijo, Célia Maria Ferreira; Cruz, Israel; Melo, Maria Norma; Silva, Aristóbolo M.

doi:10.1590/s0074-02762010000700009

Cited by 42 publications

(30 citation statements)

References 14 publications

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“…To evaluate the quality of DNA samples, each spleen DNA was tested by PCR for the mammal irbp ("interphotoreceptor retinoid binding protein") gene as an internal control (Ferreira et al, 2010). Samples negative for the abovementioned PCR protocol were tested for another internal control by PCR targeting the gapdh (Birkenheuer et al, 2003).…”

Section: Evaluation Of Dna Extraction Qualitymentioning

confidence: 99%

Assessment of a quantitative 5′ nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil

Benevenute

Dumler

Ogrzewalska

et al. 2017

Ticks and Tick-borne Diseases

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New genotypes of Anaplasmataceae agents have been detected in wild carnivores, birds and deer in Brazil. The present work aimed to investigate the presence of Ehrlichia and Anaplasma species in rodents sampled in Brazil. Additionally, a newly designed quantitative 5' nuclease real-time multiplex PCR for Ehrlichia and Anaplasma spp. detection based on groEL gene amplification was designed, showing high specificity and sensitivity (10 groEL fragment copy/μL). Between 2000 and 2011, different rodent species [n=60] were trapped in 5 Brazilian biomes. Among 458 rodent spleen samples, 0.4% (2/458) and 2.4% (11/458) were positive for Ehrlichia and Anaplasma spp., respectively. Of 458 samples, 2.0% (9/458) and 1.1% (5/458) were positive for Anaplasma sp. and Ehrlichia sp., respectively, using conventional 16S rRNA PCR assays. Maximum Likelihood phylogenetic analyse based on a small region of 16S rRNA genes positioned the Anaplasma genotypes in rodents near Anaplasma phagocytophilum or Anaplasma marginale and Anaplasma odocoilei isolates. Ehrlichia genotypes were closely related to E. canis. There was a low occurrence of Anaplasma and Ehrlichia in wild and synanthropic rodents in Brazil, suggesting the circulation of new genotypes of these agents in rodents in the studied areas.

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Section: Evaluation Of Dna Extraction Qualitymentioning

confidence: 99%

Assessment of a quantitative 5′ nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil

Benevenute

Dumler

Ogrzewalska

et al. 2017

Ticks and Tick-borne Diseases

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“…In order to evaluate the quality of DNA samples, each sample of spleen-extracted DNA was used as a template in an internal control PCR targeting the mammalian IRBP (interphotoreceptor retinoid binding protein) gene (28). Three microliters of DNA was used as a template in 25-l reaction mixtures containing 10ϫ PCR buffer, 1.0 mM MgCl 2 , 0.6 mM deoxynucleotide triphosphate (dNTPs) mixture, 1.5 U of Taq DNA polymerase (Life Technologies), 1.25 l of dimethyl sulfoxide (DMSO) (Sigma-Aldrich), and 0.5 M IRBPfwd (5=-TCCAACACCACCACTGAGATCTGGAC-3=) and IRBPrev (5=-GTG AGGAAGAAATCGGACTGGCC-3=) primers.…”

Section: Methodsmentioning

confidence: 99%

Association of Bartonella Species with Wild and Synanthropic Rodents in Different Brazilian Biomes

Gonçalves

Favacho

Roque

et al. 2016

Appl Environ Microbiol

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Bartonella spp. comprise an ecologically successful group of microorganisms that infect erythrocytes and have adapted to different hosts, which include a wide range of mammals, besides humans. Rodents are reservoirs of about two-thirds of Bartonella spp. described to date; and some of them have been implicated as causative agents of human diseases. In our study, we performed molecular and phylogenetic analyses of Bartonella spp. infecting wild rodents from five different Brazilian biomes. In order to characterize the genetic diversity of Bartonella spp., we performed a robust analysis based on three target genes, followed by sequencing, Bayesian inference, and maximum likelihood analysis. Bartonella spp. were detected in 25.6% (117/457) of rodent spleen samples analyzed, and this occurrence varied among different biomes. The diversity analysis of gltA sequences showed the presence of 15 different haplotypes. Analysis of the phylogenetic relationship of gltA sequences performed by Bayesian inference and maximum likelihood showed that the Bartonella species detected in rodents from Brazil was closely related to the phylogenetic group A detected in other cricetid rodents from North America, probably constituting only one species. Last, the Bartonella species genogroup identified in the present study formed a monophyletic group that included Bartonella samples from seven different rodent species distributed in three distinct biomes. In conclusion, our study showed that the occurrence of Bartonella bacteria in rodents is much more frequent and widespread than previously recognized. IMPORTANCEIn the present study, we reported the occurrence of Bartonella spp. in some sites in Brazil. The identification and understanding of the distribution of this important group of bacteria may allow the Brazilian authorities to recognize potential regions with the risk of transmission of these pathogens among wild and domestic animals and humans. In addition, our study accessed important gaps in the biology of this group of bacteria in Brazil, such as its low host specificity, high genetic diversity, and relationship with other Bartonella spp. detected in rodents trapped in America. Considering the diversity of newly discovered Bartonella species and the great ecological plasticity of these bacteria, new studies with the aim of revealing the biological aspects unknown until now are needed and must be performed around the world. In this context, the impact of Bartonella spp. associated with rodents in human health should be assessed in future studies.

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“…All blood samples used in the study were positive (100%) to bovine β-actin which indicated Unauthenticated Download Date | 5/10/18 3:04 AM that the extracted DNA samples were of good quality. Extracted DNA must be of good quality for molecular analysis particularly in epidemiological survey of pathogen of veterinary importance (Ferreira et al 2010). Bovine β-actin is so ubiquitous that it is considered a house-keeping protein and historically been used as an external standard for Western blotting assay (Karakozova et al 2006).…”

Section: Molecular Detection Of Theileria Spmentioning

confidence: 99%

Molecular detection and characterization of Theileria species in the Philippines

Belotindos

Lazaro

Villanueva

et al. 2014

Acta Parasitologica

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Theileriosis is a tick-borne disease of domestic and wild animals that cause devastating economic loss in livestock in tropical and subtropical regions. Theileriosis is not yet documented in the Philippines as compared to babesiosis and anaplasmosis which are considered major tick-borne diseases that infect livestock in the country and contribute major losses to the livestock industry. The study was aimed to detect Theileria sp. at genus level in blood samples of cattle using polymerase chain reaction (PCR) assay. Specifically, it determined the phylogenetic relationship of Theileria species affecting cattle in the Philippines to other Theileria sp. registered in the GenBank. A total of 292 blood samples of cattle that were collected from various provinces were used. Theileria sp. was detected in 43/292 from the cattle blood samples using PCR assay targeting the major piroplasm surface protein (MPSP) gene. DNA sequence showed high similarity (90-99%) among the reported Theileria sp. isolates in the GenBank and the Philippine isolates of Theileria. Phylogenetic tree construction using nucleotide sequence classified the Philippine isolates of Theileria as benign. However, nucleotide polymorphism was observed in the new isolate based on nucleotide sequence alignment. It revealed that the new isolate can be a new species of Theileria.

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Alternative PCR protocol using a single primer set for assessing DNA quality in several tissues from a large variety of mammalian species living in areas endemic for leishmaniasis

Cited by 42 publications

References 14 publications

Assessment of a quantitative 5′ nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil

Assessment of a quantitative 5′ nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil

Association of Bartonella Species with Wild and Synanthropic Rodents in Different Brazilian Biomes

Molecular detection and characterization of Theileria species in the Philippines

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