MARCIA DALASTRA LAURENTI

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Departamento de Patologia, Faculdade de Medicina - Docente
LIM/50 - Laboratório de Patologia das Moléstias Infecciosas, Hospital das Clínicas, Faculdade de Medicina - Líder

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  • article 22 Citação(ões) na Scopus
    Value of the oral swab for the molecular diagnosis of dogs in different stages of infection with Leishmania infantum
    (2016) ASCHAR, Mariana; OLIVEIRA, Eveline Tozzi Braga de; LAURENTI, Marcia Dalastra; MARCONDES, Mary; TOLEZANO, Jose Eduardo; HIRAMOTO, Roberto Mitsuyoshi; CORBETT, Carlos Eduardo P.; MATTA, Vania Lucia Ribeiro da
    This study was based on the need to employ a sensitive and specific method with samples that could be easily collected for diagnosing dogs infected with Leishmania infantum. To this end, we used real time-PCR (qPCR) to assess the value of the oral swab (OS) in detecting infected sick dogs (SD; n = 62), including, for the first time, the analysis of apparently healthy infected dogs (AD; n =30), both from endemic areas for visceral leishmaniasis (VL). For comparison, we also evaluated the performance of the conjunctival swab (CS), blood (BL), lymph node (LN) and serology. We detected the presence of Leishmania DNA in the oral cavity in 62 out of the 92 dogs studied. The OS positivity (67.4%) was equivalent to the CS (68.5%) (p > 0.05), higher than BL (52.2%) (p <= 0.05), and lower than LN (84.8%) (p <= 0.05). OS and CS performed well in SD dogs (82.3% and 83.9%, respectively) but not in AD dogs (36.7% for both samples). BL showed the lowest positivity (52.2%) and provided equivalent results between AD (60.0%) and SD (48.4%) dogs (p > 0.05). LN yielded the highest positivity (84.8%), and it was also higher in the SD population (93.5%) compared to the AD population (66.7%) (p <= 0.05). Parasite load was high in LN, moderate in OS and CS, and low in BL, showing the relationship between the levels of parasitism and the positivity rates found in these samples. Serology was positive in 82.2% of the SD group and in 70% of the AD dogs (p > 0.05). Among the 20 seronegative dogs, seven (35%) were positive in either OS or CS, and 12 (60%) were positive when both noninvasive samples were jointly considered. The OS/CS combination resulted in a significant increase of positivity (p <= 0.05) for the AD dogs (from 36.7% to 63.4%), as well as OS/serology (80%) and OS/CS/serology (83.4%). For the SD population, positivity reached up to 95.2% with the same combinations, showing that combination of samples and/or tests is required for the identification of dogs infected with L. infantum and that the OS and CS combination based on qPCR notably improves the detection of both AD and SD dogs. In conclusion, OS proved to be a suitable sample for the molecular diagnosis of infected dogs with clinical signs of VL, but not for dogs with inapparent infection. For these, we recommend the combination of OS results with CS and/or serology in order to reach relevant positivity for L infantum. Finally, another advantage of using OS or both noninvasive samples is the increased likelihood of diagnosing seronegative dogs.
  • article 83 Citação(ões) na Scopus
    Coinfection of Leishmania chagasi with Toxoplasma gondii, Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV) in cats from an endemic area of zoonotic visceral leishmaniasis
    (2012) VICENTE SOBRINHO, Ludmila Silva; ROSSI, Claudio Nazaretian; VIDES, Juliana Peloi; BRAGA, Eveline Tozzi; GOMES, Ana Amelia Domingues; LIMA, Valeria Marcal Felix de; PERRI, Silvia Helena Venturoli; GENEROSO, Diego; LANGONI, Helio; LEUTENEGGER, Christian; BIONDO, Alexander Welker; LAURENTI, Marcia Dalastra; MARCONDES, Mary
    The aim of the present study was to determine the coinfection of Leishmania sp. with Toxoplasma gondii, Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV) in a population of cats from an endemic area for zoonotic visceral leishmaniasis. An overall 66/302 (21.85%) cats were found positive for Leishmania sp., with infection determined by direct parasitological examination in 30/302(9.93%), by serology in 46/302(15.23%) and by both in 10/302 (3.31%) cats. Real time PCR followed by amplicon sequencing successfully confirmed Leishmania infantum (syn Leishmania chagasi) infection. Out of the Leishmania infected cats, coinfection with FIV was observed in 12/66(18.18%), with T. gondii in 17/66 (25.75%) and with both agents in 5/66(7.58%) cats. FeLV was found only in a single adult cat with no Leishmania infection. A positive association was observed in coinfection of Leishmania and FIV (p < 0.0001), but not with T. gondii (p > 0.05). In conclusion, cats living in endemic areas of visceral leishmaniasis are significantly more likely to be coinfected with Fly, which may present confounding clinical signs and therefore cats in such areas should be always carefully screened for coinfections.
  • article 6 Citação(ões) na Scopus
    Chromosomal segments may explain the antibody response cooperation for canine leishmaniasis pathogenesis
    (2020) BATISTA, Luis F. S.; TORRECILHA, Rafaela B. P.; SILVA, Rafaela B.; UTSUNOMIYA, Yuri T.; SILVA, Thais B. F.; TOMOKANE, Thaise Y.; PACHECO, Acacio D.; BOSCO, Anelise M.; PAULAN, Silvana C.; ROSSI, Claudio N.; COSTA, Gustavo N. O.; MARCONDES, Mary; CIARLINI, Paulo C.; NUNES, Caris M.; MATTA, Vania L. R.; LAURENTI, Marcia D.
    Visceral leishmaniasis (VL) is marked by hyperactivation of a humoral response secreting high quantity of immunoglobulins (Igs) that are inaccessible to intracellular parasites. Here we investigated the contributions of the antibody response to the canine leishmaniasis pathogenesis. Using correlation and genome-wide association analysis, we investigated the relationship of anti-Leishmania infantum immunoglobulin classes levels with parasite burden, clinical response, renal/hepatic biochemical, and oxidative stress markers in dogs from endemic areas of VL. Immunoglobulin G (IgG) and IgA were positively correlated with parasite burden on lymph node and blood. Increased IgG, IgA and IgE levels were associated with severe canine leishmaniasis (CanL) whereas IgM was elevated in uninfected exposed dogs. Correlations of IgM, IgG and IgA with creatinine, urea, AST and ALT levels in the serum were suggested an involvement of those Igs with renal and hepatic changes. The correlogram of oxidative radicals and antioxidants revealed a likely relationship of IgM, IgG and IgA with oxidative stress and lipid pemxidation in the blood, suggested as mechanisms mediating tissue damage and CanL worsening. The gene mapping on chromosomal segments associated with the quantitative variation of immunoglobulin classes identified genetic signatures involved with reactive oxygen species generation, phagolysosome maturation and rupture, free iron availability, Thl/Th2 differenciation and, immunoglobulin clearance. The findings demonstrated the roles of the antibody response as resistance or susceptibility markers and mediators of CanL pathogenesis. In addition we pinpointed candidate genes as potential targets for the therapy against the damage caused by exacerbated antibody response and parasitism in VL.
  • article 70 Citação(ões) na Scopus
    Leishmania chagasi infection in cats with dermatologic lesions from an endemic area of visceral leishmaniosis in Brazil
    (2011) VIDES, Juliana Peloi; SCHWARDT, Tatianna Frate; VICENTE SOBRINHO, Ludmila Silva; MARINHO, Marcia; LAURENTI, Marcia Dalastra; BIONDO, Alexander Welker; LEUTENEGGER, Christian; MARCONDES, Mary
    Although dogs are considered the main domestic reservoirs for Visceral Leishmaniosis (VL), which is caused in the Americas by Leishmania chagasi, infected cats have also been recently found in endemic areas of several countries and became a public health concern. Accordingly, the purpose of this study was to evaluate cats with dermatologic lesions from an endemic area of VL and the natural infection of L. chagasi. A total of 55 cats were selected between April 2008 and November 2009 from two major animal shelters of Aracatuba, Southeastern Brazil. All cats underwent general and dermatologic examinations, followed by direct parasitological examination of lymphoid organs, immunosorbent assay (ELISA) and indirect immunofluorescence (IFAT). In addition, detection of amastigotes was performed by immunohistochemistry (IHC) in skin lesions of all cats. VL was diagnosed in 27/55 (49.1%) cats with dermatological problems. Amastigotes were found in lymphoid organs of 10/27 (37.0%) cats; serology of 14/27 (51.9%), 6/27 (22.2%) and 5/27 (18.5%) cats was positive for ELISA, IFAT and both, respectively. The IHC identified 9/27 (33.3%) cats; 5/27 (18.5%) were positive only for IHC and therefore increased the overall sensitivity. Specific FIV antibodies were found in 6/55(10.9%) cats, of which 5/6 (83.3%) had leishmaniosis. Real time PCR followed by amplicon sequencing successfully confirmed L chagasi infection. In conclusion, dermatological lesions in cats from endemic areas was highly associated to visceral leishmaniosis, and therefore skin IHC and differential diagnosis of LV should be always conducted in dermatological patients in such areas.
  • article 60 Citação(ões) na Scopus
    Comparative evaluation of the DPP (R) CVL rapid test for canine serodiagnosis in area of visceral leishmaniasis
    (2014) LAURENTI, M. D.; LEANDRO JR., M. V. de Santana; TOMOKANE, T. Y.; LUCCA, H. R. L. De; ASCHAR, M.; SOUZA, C. S. F.; SILVA, R. M.; MARCONDES, M.; MATTA, V. L. R. da
    We investigated the performance of the DPP (R) canine visceral leishmaniasis (CVL) rapid test, a novel immunochromatographic assay launched by BioManguinhos (Brazil), which was recently included in the new Brazilian protocol for screening CVL in serological surveys. The present study compared the DPP (R) with the ELISA and IFA produced by BioManguinhos (Brazil) both with L. major-like antigens and with in-house tests using Leishmania infantum chagasi (in-house ELISA and in-house IFA). We analyzed the sera from clinically symptomatic (n = 47) and asymptomatic (n = 38) infected dogs from an endemic area of CVL, as well as from healthy (n = 18) dogs, in addition to the sera of dogs (n = 81) infected with other pathogens. The DPP (R) and the in-house ELISA showed a sensitivity of 90.6% and 94.1%, respectively, and specificity of 95.1% and 97.5%, respectively, and both presented cross-reactivity only with the sera of dogs with babesiosis, 44% for the DPP (R) and 22% for the in-house ELISA. The clinical groups were detected equally by the two assays. The ELISA BioManguinhos, IFA BioManguinhos, and in house-IFA showed a good sensitivity, 90.6%, 96.5% and 89.4%, respectively, but very low specificity, 77.8%, 69.1% and 65.8%, respectively, due to the high cross-reactivity with the sera from the animals harboring other pathogens. The in-house ELISA provided the highest accuracy (95.8%), followed by the DPP (R) (92.7%), ELISA BioManguinhos (84.3%), IFA BioManguinhos (83.1%), and in-house IFA (78.0%). The simultaneous use of the DPP (R) and ELISA BioManguinhos reached a sensitivity of 99.1% and 82.1% when used sequentially. In conclusion, the DPP (R) performed well as serological test for CVL, and detected both asymptomatic and symptomatic dogs in equal proportions. Although its sensitivity is not ideal yet, discarding the IFA and including the DPP (R) improved the accuracy of the new Brazilian CVL diagnostic protocol, particularly of detecting truly infected dogs. Moreover, considering the higher specificity of DPP (R) (95.1% vs 77.8%), positive predictive value (95.1% vs 81.1%) and positive likelihood value (18.3% vs 4.1%) in comparison with the ELISA BioManguinhos, the use of DPP (R) as a confirmatory test instead of a screening test is suggested.
  • article 1 Citação(ões) na Scopus
  • article 123 Citação(ões) na Scopus
    Asymptomatic dogs are highly competent to transmit Leishmania (Leishmania) infantum chagasi to the natural vector
    (2013) LAURENTI, Marcia Dalastra; ROSSI, Claudio Nazaretian; MATTA, Vania Lucia Ribeiro da; TOMOKANE, Thaise Yumie; CORBETT, Carlos Eduardo Pereira; SECUNDINO, Nagila Francinete Costa; PIMENTA, Paulo Filemon Paulocci; MARCONDES, Mary
    We evaluated the ability of dogs naturally infected with Leishmania (Leishmania) infantum chagasi to transfer the parasite to the vector and the factors associated with transmission. Thirty-eight infected dogs were confirmed to be infected by direct observation of Leishmania in lymph node smears. Dogs were grouped according to external clinical signs and laboratory data into symptomatic (n = 24) and asymptomatic (n = 14) animals. All dogs were sedated and submitted to xenodiagnosis with F1-laboratory-reared Lutzomyia longipalpis. After blood digestion, sand flies were dissected and examined for the presence of promastigotes. Following canine euthanasia, fragments of skin, lymph nodes, and spleen were collected and processed using immunohistochemistry to evaluate tissue parasitism. Specific antibodies were detected using an enzyme-linked immunosorbent assay. Antibody levels were found to be higher in symptomatic dogs compared to asymptomatic dogs (p = 0.0396). Both groups presented amastigotes in lymph nodes, while skin parasitism was observed in only 583% of symptomatic and in 35.7% of asymptomatic dogs. Parasites were visualized in the spleens of 66.7% and 71.4% of symptomatic and asymptomatic dogs, respectively. Parasite load varied from mild to intense, and was not significantly different between groups. All asymptomatic dogs except for one (93%) were competent to transmit Leishmania to the vector, including eight (61.5%) without skin parasitism. Sixteen symptomatic animals (67%) infected sand flies; six (37.5%) showed no amastigotes in the skin. Skin parasitism was not crucial for the ability to infect Lutzomyia longipalpis but the presence of Leishmania in lymph nodes was significantly related to a positive xenodiagnosis. Additionally, a higher proportion of infected vectors that fed on asymptomatic dogs was observed (p = 0.0494). Clinical severity was inversely correlated with the infection rate of sand flies (p = 0.027) and was directly correlated with antibody levels (p = 0.0379). Age and gender did not influence the transmissibility. Our data show that asymptomatic dogs are highly infective and competent for establishing sand fly infection, indicating their role in maintaining L. (L.) infantum chagasi cycle as well as their involvement in VL spreading in endemic areas.