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| Monte,Leonardo Garcia; Leal,Fernanda Munhoz Dos Anjos; Hartwig,Daiane Drawanz; Vasconcellos,Sílvio Arruda; Brihuega,Bibiana; Dellagostin,Odir Antonio; Hartleben,Cláudia Pinho. |
| The production of recombinant LipL32 protein using Escherichia coli has been used extensively for the development of vaccines and diagnostic tests for leptospirosis. However, E. coli has demonstrated limitations, including low yield and lack of post-translational modifications. In this study, rLipL32 was produced in eukaryotic expression system (Pichia pastoris) and evaluated the antigen by enzyme-linked immunosorbent assay (ELISA). The yield obtained from the culture supernatant reached 270 mg/L and ELISA showed an accuracy of 95.34%. In summary, the production of rLipL32 using P. pastoris did not impair the antigenic characteristics of this antigen and ensured its use for detecting the leptospiral antibodies in swine sera. |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Pichia pastoris; LipL32; ELISA; Leptospirosis. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000300007 |
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| Lüdtke,Charli Beatriz; Coutinho,Mariana Loner; Jouglard,Sandra Denise Dorneles; Moreira,Cecília Nunes; Fernandes,Cláudia Hartleben Pinho; Brod,Claudiomar Soares; Haake,David A.; Ko,Albert Icksang; Dellagostin,Odir Antonio; Aleixo,José Antonio Guimarães. |
| Two hybridomas secreting monoclonal antibodies (MAbs) that react with a lipoprotein (LipL32) of the outer membrane of pathogenic Leptospira were obtained. For hybridoma production, spleen cells from BALB/c mice imunized with recombinant LipL32 (rLipL32) were fused to SP2/O-Ag14 cells, selected in HAT medium and screened in an indirect ELISA. One MAb produced was of the IgG2b isotype and the other was an IgM. MAbs specificity was confirmed by indirect ELISA and immunoblotting using purified rLipL32 and whole-cell antigen preparations from Escherichia coli (E. coli) expressing LipL32 and from pathogenic and non-pathogenic serovars. Both Mabs reacted with most of the pathogenic serovars tested and none reacted with non-pathogenic Leptospira. The MAbs... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Leptospirosis; Monoclonal antibodies; LipL32. |
| Ano: 2003 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000500001 |
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| Vedhagiri,Kumaresan; Natarajaseenivasan,Kalimuthusamy; Prabhakaran,Shanmugarajan G.; Selvin,Joseph; Narayanan,Ramasamy; Shouche,Yogesh S.; Vijayachari,Paluru; Ratnam,Sivalingam. |
| The main goal of this study was to evaluate the prevalence of leptospirosis among field rodents of Tiruchirappalli district, Tamil Nadu, India. In total 35 field rats were trapped and tested for seroprevalence by the microscopic agglutination test (MAT). Isolation of leptospires was performed from blood and kidney tissues and characterized to serovar level. Genomospecies identification was carried out using 16S rRNA and lipL32 gene sequencing. The molecular phylogeny was constructed to find out species segregation. Seroprevalence was about 51.4 %, and the predominant serovars were Autumnalis, Javanica, Icterohaemorrhagiae and Pomona. Two isolates from the kidneys were identified as serovar Javanica of Serogroup Javanica, and sequence based molecular... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Leptospirosis; Leptospira borgpetersenii; LipL32; 16S rRNA. |
| Ano: 2010 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000100022 |
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| Monte,Leonardo Garcia; Jorge,Sérgio; Luiz,João Paulo Mesquita; Sinnott,Francine; Seixas,Fabiana Kömmling; Aleixo,José Antonio Guimarães; Samartino,Luis Ernesto; Conceição,Fabricio Rochedo; Hartleben,Cláudia Pinho. |
| Diagnosis of leptospirosis by PCR is hampered due to the presence of substances on biological fluids. Here, we report an immunomagnetic separation step prior to PCR which improved the detection of Leptospira spp. in blood and urine samples from dogs. It resulted in a significant improvement on sensitivity for diagnosis of canine leptospirosis. |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Canine leptospirosis; IMS-PCR; LipL32. |
| Ano: 2012 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200023 |
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