Diagnosis of plague and identification of virulence markers in Yersinia pestis by multiplex-PCR

Leal, Nilma Cintra; Almeida, Alzira Maria Paiva de

doi:10.1590/s0036-46651999000600002

Cited by 41 publications

(39 citation statements)

References 18 publications

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“…In Brazil, since 2005, there have been no laboratory-confirmed human cases and the last Y. pestis isolation occurred in 1997 in the Ibiapaba focus (Ceará State); however, serological surveys continuously detect plague antibodies in sentinel animals in plague areas, suggesting that they remain active (Leal and Almeida, 1999;Aragão et al, 2009;WHO, 2006WHO, , 2009. Quiescent periods characterized by the absence or occurrence of rare human cases, may be misleading, since plague may reappear after many apparent control decades and its spreading potential by international trade or bioterrorism must be considered (Duplantier et al, 2005;Gage and Kosoy, 2005;WHO, 2006WHO, , 2009.…”

Section: Discussionmentioning

confidence: 99%

Subtyping Brazilian Yersinia pestis strains by pulsed-field gel electrophoresis

Barros

Silveira-Filho²,

Lins³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. We subtyped Brazilian Yersinia pestis strains by pulsed-field gel electrophoresis (PFGE). This was done with 22 Brazilian Y. pestis strains: 17 from an outbreak and 5 from endemic routine surveillance. The strains were divided into 2 groups (I and II), 8 subgroups (A-H) and 19 PFGE profiles or pulsotypes. PFGE did not separate outbreak from non-outbreak strains, as identical pulsotype patterns were found among outbreak strains and strains obtained from surveillance. However, it was able to detect intraspecific genetic diversity among Brazilian strains. This PFGE technique was able to differentiate a homogeneous group of Brazilian Y. pestis strains.

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Section: Discussionmentioning

confidence: 99%

Subtyping Brazilian Yersinia pestis strains by pulsed-field gel electrophoresis

Barros

Silveira-Filho²,

Lins³

et al. 2013

Genet. Mol. Res.

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show abstract

“…The occurrence of new cases has been rare, and the last cases were in 1997 and 2005 in Ceará. However, serological surveys continuously detect plague antibodies in sentinel animals in plague areas (Leal and Almeida, 1999;Aragão et al, 2009), suggesting that these plague foci remain active and may reappear (Duplantier et al, 2005;Gage and Kosoy, 2005;Stenseth et al, 2008;WHO, 2006WHO, , 2009.…”

Section: Discussionmentioning

confidence: 99%

Genetic diversity of Yersinia pestis in Brazil

Oliveira¹,

Barros²,

Silveira-Filho³

et al. 2012

Genet. Mol. Res.

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ABSTRACT. Plague outbreaks are occasionally reported in Brazil. Unfortunately, due to great genetic similarity, molecular subtyping of Yersinia pestis strains is difficult. Analysis of multiple-locus variable number of tandem repeats (VNTR), also known as MLVA, has been found to be a valuable tool to discriminate among strains. To check for genetic differences, strains obtained from two different ecological complexes in Brazil collected during two different epidemiological events, an epizootic in Sítio Alagoinha in 1967 and an outbreak in Planalto da Borborema in 1986, were subtyped through MLVA using 12 VNTR loci. Three clusters (A, B and C) were observed. Of the 20 strains from the epizootic, 18 fit into cluster A. Cluster A was divided into two subgroups: A 1 (15 strains) and A 2 (3 strains). Of the 17 strains from the outbreak, 15 fit into cluster B. Cluster B was divided into three subgroups: B 1 (4 strains), B 2 (4 strains) and B 3 (7 strains). Cluster C is a singleton with one epizootic strain. The external standards, Y. pestis CO92 and Y. pseudotuberculosis IP32953, formed two clusters of singletons. The stability of 12 VNTR loci of three unrelated cultures ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 11 (3): 3414-3424 (2012) Brazilian Yersinia pestis diversity 3415 included in this study was assessed. The 12 VNTR loci were stable through multiple serial subcultures in the laboratory. MLVA revealed that Y. pestis populations in Brazil are not monomorphic, and that there is intraspecific genetic diversity among Brazilian plague strains. We conclude that there is some correlation among genetic groups of this species, related to the temporal and geographic origin of isolates.

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“…After criofracture, the organ samples (about 300 mg) were submitted to a pre-treatment, 23,24 before digestion with Proteinase-K. Thus, after maceration with liquid nitrogen (N2 liq), the material was transferred to 1.5 mL micro tube with 600 μL of TrisHCl 10mM; EDTA 1mM; pH8,0 (TE).…”

Section: Molecular Methodsmentioning

confidence: 99%

Search for Mycobacterium leprae in wild mammals

Pedrini¹,

Rosa²,

Medri³

et al. 2010

Braz J Infect Dis

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Leprosy is still a worldwide public health problem. Brazil and India show the highest prevalence rates of the disease. Natural infection of armadillos Dasypus novemcinctus with Mycobacterium leprae has been reported in some regions of the United States. Identification of bacilli is difficult, particularly due to its inability to grow in vitro. The use of molecular tools represents a fast and sensitive alternative method for diagnosis of mycobacteriosis. In the present study, the diagnostic methods used were bacilloscopy, histopathology, microbiology, and PCR using specific primers for M. leprae repetitive sequences. PCR were performed using genomic DNA extracted from 138 samples of liver, spleen, lymph nodes, and skin of 44 D. novemcinctus, Euphractus sexcinctus, Cabassous unicinctus, and C. tatouay armadillos from the Middle Western region of the state of São Paulo and from the experimental station of Embrapa Pantanal, located in Pantanal da Nhecolândia of Mato Grosso do Sul state. Also, the molecular analysis of 19 samples from internal organs of other road killed species of wild animals, such as Nasua nasua (ring-tailed coati), Procyon cancrivoros (hand-skinned), Cerdocyon thous (dog-pity-bush), Cavia aperea (restless cavy), Didelphis albiventris (skunk), Sphigurrus spinosus (hedgehog), and Gallictis vittata (ferret) showed PCR negative data. None of the 157 analyzed samples had shown natural mycobacterial infection. Only the armadillo inoculated with material collected from untreated multibacillary leprosy patient presented PCR positive and its genomic sequencing revealed 100% identity with M. leprae. According to these preliminary studies, based on the used methodology, it is possible to conclude that wild mammals seem not to play an important role in the epidemiology of leprosy in the Middle Western region of the São Paulo state and in the Pantanal of Mato Grosso do Sul state.

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Diagnosis of plague and identification of virulence markers in Yersinia pestis by multiplex-PCR

Cited by 41 publications

References 18 publications

Subtyping Brazilian Yersinia pestis strains by pulsed-field gel electrophoresis

Subtyping Brazilian Yersinia pestis strains by pulsed-field gel electrophoresis

Genetic diversity of Yersinia pestis in Brazil

Search for Mycobacterium leprae in wild mammals

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