Culture-Independent Detection and Identification of
            <i>Leptospira</i>
            Serovars

Matthias, Michael; Lubar, Aristea; Acharige, Shalka S Lanka; Chaiboonma, Kira; Pilau, Nicholas Nathaniel; Marroquin, Alan S; Jayasundara, Dinesha; Agampodi, Suneth; Vinetz, Joseph M.

doi:10.1128/spectrum.02475-22

Cited by 4 publications

(2 citation statements)

References 38 publications

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“…Both sets of serovars can be used together, with the new serovars being utilised for more accurate and quicker diagnosis. This development offers a chance to enhance the comprehension and control of leptospirosis by improving diagnostic precision and epidemiological monitoring [ 83 , 84 , 85 , 86 , 87 , 88 ].…”

Section: Discussionmentioning

confidence: 99%

Improved Antibody Detection for Canine Leptospirosis: ELISAs Modified Using Local Leptospiral Serovar Isolates from Asymptomatic Dogs

Boonciew,

Saisongkorh,

Brameld

et al. 2024

Animals

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Leptospirosis is a zoonotic disease of significant concern for human and animal health, with domestic animals, including dogs, acting as reservoirs for human infection. Serology is widely used for leptospirosis diagnosis, even though the standard microscopic agglutination test (MAT) using a panel of serovars lacks specificity and can lead to detection limitations in certain regions. In this study, we aimed to develop an antibody detection tool for dogs using an indirect enzyme-linked immunosorbent assay (ELISA) with a set of local serovar isolates, including Paidjan, Dadas, and Mini, to enhance the accuracy of leptospirosis surveillance in our region. The specificity and sensitivity of various antigen preparations, namely leptospiral whole-cell protein (WCP), total membrane protein (TMP), and outer membrane protein (OMP), were assessed using sera from infected and non-infected dogs, as well as negative puppy sera. Leptospirosis diagnosis was supported using a genus-specific nested polymerase chain reaction test on all collected sera. Protein preparations were validated using SDS-PAGE and Western blotting analysis. In the results, the standard MAT failed to detect antibodies in any of the dogs confirmed as being infected using PCR and isolation, highlighting its limitations. In contrast, the OMP-based ELISAs using local isolates of Leptospira serovars gave positive results with sera from all infected dogs, and negative results with sera from all dogs from non-endemic areas. IgG titres of infected and unvaccinated dogs from endemically affected areas were significantly higher than those in non-endemic regions. Using the OMP-based IgG/ELISAs with the local serovar Dadas resulted in higher specificity and lower sensitivity than when using the WCP- and TMP-based IgG/ELISAs. Agreement analysis revealed fair and moderate concordance between OMP-based IgG/ELISAs and PCR results, whereas slight and fair agreement was observed between OMP-based ELISAs and the MAT. Overall, the modified OMP-based IgG/ELISAs, utilising relevant local serovar isolates from dogs, demonstrated improved accuracy in detecting leptospirosis in the study area, overcoming the limitations of the MAT. This study highlights the importance of identifying and incorporating these local circulating serovar isolates into serological techniques for leptospirosis diagnosis and surveillance.

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Section: Discussionmentioning

confidence: 99%

Improved Antibody Detection for Canine Leptospirosis: ELISAs Modified Using Local Leptospiral Serovar Isolates from Asymptomatic Dogs

Boonciew,

Saisongkorh,

Brameld

et al. 2024

Animals

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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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Complete genome sequences of twelve strains of Leptospira interrogans isolated from humans in Sri Lanka

Senavirathna

Jayasundara

Warnasekara

et al. 2023

Infection, Genetics and Evolution

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Culture-Independent Detection and Identification of Leptospira Serovars

Abstract: Leptospirosis is caused by a diverse group of pathogenic spirochetes comprising over 200 different serotypes. Some are widely reported and of public health importance owing to their propensity to produce severe, fatal disease in humans.

Cited by 4 publications

References 38 publications

Improved Antibody Detection for Canine Leptospirosis: ELISAs Modified Using Local Leptospiral Serovar Isolates from Asymptomatic Dogs

Improved Antibody Detection for Canine Leptospirosis: ELISAs Modified Using Local Leptospiral Serovar Isolates from Asymptomatic Dogs

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

Complete genome sequences of twelve strains of Leptospira interrogans isolated from humans in Sri Lanka

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