Genetic Analysis of LigA, LipL32, Loa22, and OmpL1 Antigens from Leptospira spp. Sejroe Serogroup: How Close We Are To a Vaccine Against Bovine Leptospirosis?

Azevedo, Maria Isabel Nogueira Di; Borges, Ana Luiza dos Santos Baptista; Kremer, Frederico Schmitt; Melo, Juliana dos Santos Loria de; Carvalho-Costa, Filipe Aníbal; Lilenbaum, Walter

doi:10.1007/s00284-023-03414-w

Cited by 3 publications

(2 citation statements)

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“…Therefore, this protein can be used in the development of promising diagnostic tools [ 33 ]. The LipL32 protein is a highly conserved Leptospira -specific antigen [ 34 ] and has great diagnostic potential for the detection of leptospirosis by ELISA [ 35 ]. In this study, rLipL32 protein production was performed to produce and characterize IgG anti-rLipL32 as an antigen source for developing leptospirosis diagnostic techniques.…”

Section: Discussionmentioning

confidence: 99%

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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Background and Aim: Microscopic agglutination test (MAT) for the diagnosis of leptospirosis requires live cultures and is serovar-specific, while polymerase chain reaction (PCR) requires expensive equipment and sample preparation. The rLipL32 protein is conserved and can be used for the production of immunoglobulin G (IgG) anti-rLipL32 antibody, which can be used as a biomarker for leptospirosis diagnosis. This study aimed to produce and characterize an IgG anti-rLipL32 antibody as a biomarker for leptospirosis diagnosis. Materials and Methods: Escherichia coli rLipL32 was cultured and analyzed by PCR and sequencing. Cultures were used for rLipL32 protein expression and purification and the rLipL32 protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The rLipL32 protein was used to produce anti-rLipL32 serum and was analyzed by enzyme-linked immunosorbent assay (ELISA). Serum was purified to obtain IgG anti-rLipL32 antibody and characterized by SDS-PAGE and western blotting. Results: PCR was able to amplify the LipL32 gene from E. coli rLipL32, and sequencing analysis showed 99.19% similarity with pathogenic Leptospira. SDS-PAGE analysis showed a 32-kDa band. ELISA results showed an increase in OD in anti-rLipL32 serum compared to preimmune serum. Western blotting results showed that the IgG anti-rLipL32 antibody was able to bind and cross-reacts with pathogenic Leptospira serovar but not with E. coli or Staphylococcus aureus. Conclusion: IgG anti-rLipL32 antibody has high specificity and sensitivity against Leptospira pathogens. These findings suggest that IgG anti-rLipL32 antibody is a promising biomarker for the diagnosis of leptospirosis. Keywords: anti-rLipL32 serum, immunoglobulin G anti-rLipL32 antibody, Leptospira, rLipL32 protein.

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Section: Discussionmentioning

confidence: 99%

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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“…Thirteen primer sets were either modified from existing primer sets (60,(88)(89)(90)(91)(92)(93)(94)(95)(96)(97)(98)…”

Section: Ampseq Methodsmentioning

confidence: 99%

Host population structure and rare dispersal events drive leptospirosis transmission patterns amongRattus norvegicusin Boston, Massachusetts, US

Stone,

Hamond,

Clegg

et al. 2024

Preprint

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Leptospirosis (caused by pathogenic bacteria in the genusLeptospira) is prevalent worldwide but more common in tropical and subtropical regions. Transmission can occur following direct exposure to infected urine from reservoir hosts, such as rats, or a urine-contaminated environment, which then can serve as an infection source for additional rats and other mammals, including humans. The brown rat,Rattus norvegicus, is an important reservoir of leptospirosis in urban settings. We investigated leptospirosis among brown rats in Boston, Massachusetts and hypothesized that rat dispersal in this urban setting influences the movement, persistence, and diversity ofLeptospira. We analyzed DNA from 328 rat kidney samples collected from 17 sites in Boston over a seven-year period (2016–2022); 59 rats representing 12 of 17 sites were positive forLeptospira. We used 21 neutral microsatellite loci to genotype 311 rats and utilized the resulting data to investigate genetic connectivity among sampling sites. We generated whole genome sequences for 28Leptospiraisolates obtained from frozen and fresh tissue from some of the 59Leptospira-positive rat kidneys. When isolates were not obtained, we attemptedLeptospiragenomic DNA capture and enrichment, which yielded 14 additionalLeptospiragenomes from rats. We also generated an enrichedLeptospiragenome from a 2018 human case in Boston. We found evidence of high genetic structure and limited dispersal among rat populations that is likely influenced by major roads and/or other unknown dispersal barriers, resulting in distinct rat population groups within the city; at certain sites these groups persisted for multiple years. We identified multiple distinct phylogenetic clades ofL. interrogansamong rats, with specific clades tightly linked to distinct rat populations. This pattern suggestsL. interroganspersists in local rat populations and movement of leptospirosis in this urban rat community is driven by rat dispersal. Finally, our genomic analyses of the 2018 human leptospirosis case in Boston suggests a link to rats as the source. These findings will be useful for guiding rat control and human leptospirosis mitigation efforts in this and other urban settings.

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Monitoring the spread of leptospirosis agent as one of the reasons of low-quality milk

Zaritskyi,

Zhuk,

Kovpak

et al. 2023

Potr. S. J. F. Sci.

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On the global scale of the zoonoses problem, leptospirosis is among the five diseases that pose the greatest threat to humankind today. Leptospirosis is a worldwide zoonotic disease caused by pathogenic Leptospira species. In general, leptospirosis has been registered in more than 150 species of mammals. There are about 300 serovars of Leptospira spp. Serovar Hardjo is one of the most common causes of leptospirosis among cattle globally. In cows, the infection can be completely asymptomatic or cause abortions, stillbirths, infertility, and mastitis. The study's relevance is determined by the negative impact on the economy – productivity loss, and high cost of medical-preventive activities. Leptospirosis also affects humans. In this regard, the present study aimed to determine the prevalence of antibodies to Leptospira interrogans serovar Hardjo in tank milk samples from cows selected from farms in different regions of Ukraine. The method of indirect enzyme-multiplied immunoassay was used for this problem to be investigated. We have investigated 114 tank samples from 66 Ukrainian farms, 63.2% positive, and 36.8% negative. It was established that antibodies to the causative agent of leptospirosis were recorded in different regions of Ukraine. It has been established that the largest number of positive samples was from Kyiv and Cherkasy regions. Our study results complement the study results of other authors and indicate the circulation of this causative agent among the cows in Ukraine, as well as being of practical value for diagnosing and controlling leptospirosis among the cattle.

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Genetic Analysis of LigA, LipL32, Loa22, and OmpL1 Antigens from Leptospira spp. Sejroe Serogroup: How Close We Are To a Vaccine Against Bovine Leptospirosis?

Cited by 3 publications

References 41 publications

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Host population structure and rare dispersal events drive leptospirosis transmission patterns amongRattus norvegicusin Boston, Massachusetts, US

Monitoring the spread of leptospirosis agent as one of the reasons of low-quality milk

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