Evaluation of Recombinant
            <i>Leptospira</i>
            Antigen-Based Enzyme-Linked Immunosorbent Assays for the Serodiagnosis of Leptospirosis

Flannery, Brendan; Costa, Dirceu Joaquim; Carvalho, Fernanda Pinheiro; Guerreiro, Hygia; Matsunaga, James; Silva, Emilson Domingos da; Ferreira, Antônio G. P.; Riley, Lee W.; Reis, Mitermayer Galvão dos; Haake, David A.; Ko, Albert I.

doi:10.1128/jcm.39.9.3303-3310.2001

Cited by 148 publications

(123 citation statements)

References 46 publications

(51 reference statements)

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“…Lsa44 and Lsa45 share serum recognition properties with the adhesins described elsewhere, LipL32 (Flannery et al, 2001;Xu et al, 2008), LigA and LigB proteins (Palaniappan et al, 2004;Srimanote et al, 2008), Lsa27 (Longhi et al, 2009), Lsa63 (Vieira et al, 2010a), Lsa66 (Oliveira et al, 2011), and OmpL37 (Pinne et al, 2010), which have shown positive reactivity with serum samples from patients diagnosed with leptospirosis. Protein recognition by serum (1), or lsPBS containing PBS-(2), Lsa44-(3) or Lsa45-injected mouse serum (4), at a dilution of 1 : 100.…”

Section: Discussionmentioning

confidence: 99%

Functional and immunological evaluation of two novel proteins of Leptospira spp.

et al. 2014

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This work shows the production and characterization of two novel putative lipoproteins encoded by the genes LIC10645 and LIC10731 identified in the genome sequences of Leptospira interrogans. In silico conservation analysis indicated that the proteins are well conserved among pathogenic leptospiral serovars and species. Recombinant proteins were obtained in Escherichia coli BL21(DE3) Star pLysS strain, purified by metal-affinity chromatography, and used for characterization and immunological evaluations. Recombinant proteins were capable of eliciting a combination of humoral and cellular immune responses in animal models, and could be recognized by antibodies present in human serum samples. The recombinant proteins Lsa44 and Lsa45 were able to bind laminin, and were named Lsa44 and Lsa45 for leptospiral surface adhesins of 44 and 45 kDa, respectively. The attachment to laminin was dose-responsive with K D values of 108.21 and 250.38 nM for Lsa44 and Lsa45, respectively. Moreover, these proteins interact with plasminogen (PLG) with K D values of 53.56 and 36.80 nM, respectively. PLG bound to the recombinant proteins could be converted to plasmin (PLA) in the presence of an activator. Cellular localization assays suggested that the Lsa44 and Lsa45 were surface-exposed. These are versatile proteins capable of interacting with laminin and PLG/PLA, and hence could mediate bacterial adhesion and contribute to tissue penetration. INTRODUCTIONLeptospirosis is a zoonosis of global importance caused by pathogenic bacterial species of the genus Leptospira (Bharti et al., 2003;Faine et al., 1999). In urban environments, rodents are the main host reservoirs of leptospires, shedding live bacteria through their urine (Ko et al., 1999;Vinetz et al., 1996). Humans are infected via contact with urine of wild or domestic animal carriers, either directly or indirectly through contaminated water or soil (Adler & de la Peña Moctezuma, 2010). The disease presents a broad spectrum of symptoms, including fever, vomiting, headache, diarrhoea, and abdominal and generalized muscle pain. Due to these flu-like signs, leptospirosis remains under-diagnosed. Progression to multiorgan system complications, known as Weil's syndrome, occurs in 5-15 % of cases, with mortality rates of 5-40 % (Faine et al., 1999;Ko et al., 1999;Levett, 2001; Plank & Dean, 2000).Currently available commercial vaccines are based on inactivated whole-cell or membrane preparations of pathogenic leptospires, named bacterins. They confer protective responses mostly through the induction of antibodies against LPS antigens (Adler & de la Peña Moctezuma, 2010; de la Peña-Moctezuma et al., 1999). Nevertheless, these vaccines do not induce long-term protection against infection and do not provide cross-protective immunity against leptospiral serovars not included in the vaccine preparation (Adler & de la Peña Moctezuma, 2010). Due to the large number of serovars (Bharti et al., 2003), the search for conserved and protective antigens is being pursued (Koizumi & Watanabe, 2005...

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Section: Discussionmentioning

confidence: 99%

Functional and immunological evaluation of two novel proteins of Leptospira spp.

et al. 2014

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“…Recombinant protein-based serologic tests present high sensitivity and specificity related to the high concentration of immunoreactive antigens which can be used in assays and the lack of nonspecific moieties present in whole-cell preparations (16). ELISA for leptospirosis has been used for human and animal infection diagnostic.…”

Section: Discussionmentioning

confidence: 99%

“…ELISA for leptospirosis has been used for human and animal infection diagnostic. Recombinant protein has been produced using porin transmembrane protein (OmpL1), lipoproteins (LipL32, LipL36 and LipL41) and a heat-shock protein (Hsp58), all of which have been used in ELISA for serodiagnosis of human leptospirosis (16). The LipL32 is a major leptospiral outer membrane protein whose expression is restricted to pathogenic Leptospira species and it is highly conserved within more than 250 pathogenic Leptospira serovars.…”

Section: Discussionmentioning

confidence: 99%

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Leptospirosis serosurvey in bovines from Brazilian Pantanal using IGG ELISA with recombinant protein LipL32 and microscopic agglutination test

Tomich¹,

Bomfim²,

Koury³

et al. 2007

Braz. J. Microbiol.

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This investigation was carried out in Brazilian Pantanal: region with important biodiversity. This region's climatic conditions, hydrology and geomorphology as well as the existence of great variety of wild species favor the maintenance of the Leptospira in the environment. The aim of this study was to evaluate IgG ELISA with recombinant protein LipL32 in comparison with microscopic agglutination test (MAT) and additionally contribute to the knowledge of the distribution of the one of most important worldwide zoonotic infection, assessing the seropositivity of bovine leptospirosis in beef cattle herds of Brazilian Pantanal, an important ecological preserved area, where cattle constitute not only the most important economic resource but also the major activity compatible of the conservation of natural resource of the region. Out of 282 samples of cattle serum analyzed, 143 (50.71%) were positive in MAT. The serovar Hardjo (genotypic Hardjoprajitno and Hardjobovis), Wolffi and Ballum showed the largest frequency of reactive samples. In the IgG ELISA rLipL32, 161 samples (57.09%) were positive. This result was higher than obtained by MAT (p<0.001). The sensitivity of the ELISA test was 99.30% and the specificity was 86.33%, based on the MAT. This test was shown to be a more sensitive, specific and accurate test for the diagnosis of bovine leptospirosis compared to the MAT.

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“…La determinación de IgM por ELISA o por test rápidos inmunocromatográficos permite diagnosticar infección aguda. Diversos estudios han mostrado sensibilidad variable entre 36 y 100% para ELISA y entre 52 y 94% para los métodos inmunocromatográficos [10][11][12][13][14][15][16][17] que son más sencillos, rápi-dos y no requieren equipamiento.…”

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Prevalencia y presencia de factores de riesgo de leptospirosis en una población de riesgo de la Región Metropolitana

Abarca

et al. 2005

Rev. méd. Chile

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Evaluation of Recombinant Leptospira Antigen-Based Enzyme-Linked Immunosorbent Assays for the Serodiagnosis of Leptospirosis

Cited by 148 publications

References 46 publications

Functional and immunological evaluation of two novel proteins of Leptospira spp.

Functional and immunological evaluation of two novel proteins of Leptospira spp.

Leptospirosis serosurvey in bovines from Brazilian Pantanal using IGG ELISA with recombinant protein LipL32 and microscopic agglutination test

Prevalencia y presencia de factores de riesgo de leptospirosis en una población de riesgo de la Región Metropolitana

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