A duplex endpoint PCR assay for rapid detection and differentiation of Leptospira strains

Benacer, Douadi; Zain, Siti Nursheena Mohd; Lewis, J. W.; Khalid, Mohd Khairul Nizam Mohd; Thong, Kwai Lin

doi:10.1590/0037-8682-0364-2016

Cited by 8 publications

(4 citation statements)

References 13 publications

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“…MAT is a widely used diagnostic reference method for many studies, though not accessible in many laboratories due to its cost. MAT testing has many limitations: high levels of PLOS NEGLECTED TROPICAL DISEASES detectable antibodies are needed for a positive result and usually do not occur before the fourth week after disease onset [50](50) and it is time consuming and labour intensive [51,52]. Despite these drawbacks, the MAT test remains the only gold standard serological test and is considered a reference diagnostic test for leptospirosis in many settings [6,53].…”

Section: Discussionmentioning

confidence: 99%

Epidemiology of leptospirosis in Tanzania: A review of the current status, serogroup diversity and reservoirs

Motto

Shirima²,

Bronsvoort³

et al. 2021

PLoS Negl Trop Dis

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Background Tanzania is among the tropical countries of Sub-Saharan Africa with the environmental conditions favorable for transmission of Leptospira. Leptospirosis is a neglected zoonotic disease, and although there are several published reports from Tanzania, the epidemiology, genetic diversity of Leptospira and its host range are poorly understood. Methods We conducted a comprehensive review of human and animal leptospirosis within the 26 regions of the Tanzanian mainland. Literature searches for the review were conducted in PubMed and Google Scholar. We further manually identified studies from reference lists among retrieved studies from the preliminary search. Results We identified thirty-four studies describing leptospirosis in humans (n = 16), animals (n = 14) and in both (n = 4). The number of studies varied significantly across regions. Most of the studies were conducted in Morogoro (n = 16) followed by Kilimanjaro (n = 9) and Tanga (n = 5). There were a range of study designs with cross-sectional prevalence studies (n = 18), studies on leptospirosis in febrile patients (n = 13), a case control study in cattle (n = 1) and studies identifying novel serovars (n = 2). The most utilized diagnostic tool was the microscopic agglutination test (MAT) which detected antibodies to 17 Leptospira serogroups in humans and animals. The Leptospira serogroups with the most diverse hosts were Icterohaemorrhagiae (n = 11), Grippotyphosa (n = 10), Sejroe (n = 10), Pomona (n = 9) and Ballum (n = 8). The reported prevalence of Leptospira antibodies in humans ranged from 0.3–29.9% and risk factors were associated with occupational animal contact. Many potential reservoir hosts were identified with the most common being rodents and cattle. Conclusion Leptospirosis is prevalent in humans and animals in Tanzania, although there is regional and host variation in the reports. Many regions do not have information about the disease in either humans or their animal reservoirs. More studies are required to understand human leptospirosis determinants and the role of livestock in leptospirosis transmission to humans for the development of appropriate control strategies.

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Section: Discussionmentioning

confidence: 99%

Epidemiology of leptospirosis in Tanzania: A review of the current status, serogroup diversity and reservoirs

Motto

Shirima²,

Bronsvoort³

et al. 2021

PLoS Negl Trop Dis

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“…Detection of Leptospira DNA was performed by end-point multiplex polymerase chain reaction (PCR) following the protocol previously described. 10 DNA was extracted from 0.5 ml of whole blood as previously described. 11 PCR was performed using a C1000 Touch TM Thermal Cycler (Bio-Rad Laboratories, Hercules, California.…”

Section: Detection Of Anti-leptospira Igg and Igm Antibodies And Lept...mentioning

confidence: 99%

Leptospirainfection in people in the city of Durango, Mexico: a cross sectional study

Sandoval-Carrillo¹,

Sálas-Pacheco²,

Antuna-Salcido³

et al. 2021

J Int Med Res

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Objective To determine the seroprevalence of Leptospira immunoglobulin (Ig)G and IgM antibodies and its association with the characteristics of the study population from the northern Mexican city of Durango, Mexico. Methods Through a cross-sectional study design, inhabitants of Durango City, Mexico were surveyed between June 2018 and November 2018. Serum samples from the subjects were analysed for anti- Leptospira IgG and IgM antibodies using commercially available enzyme-linked immunosorbent assays. Sociodemographic, clinical, behavioural and housing characteristics were recorded. Data were analysed by bivariate and multivariate analyses. Results The study enrolled 413 people, of which 124 (30.0%) and 137 (33.2%) were positive for anti- Leptospira IgG antibodies and anti- Leptospira IgM antibodies, respectively. Multivariate analysis showed that Leptospira seropositivity was associated with professional occupation, alcohol consumption, ill clinical status, memory impairment and a history of surgery. Conclusions This is the first study to report the seroepidemiology of Leptospira infection in an urban general population in the north of Mexico. The seroprevalence of Leptospira infection found was higher than those previously reported in Mexican studies.

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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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A duplex endpoint PCR assay for rapid detection and differentiation of Leptospira strains

Cited by 8 publications

References 13 publications

Epidemiology of leptospirosis in Tanzania: A review of the current status, serogroup diversity and reservoirs

Epidemiology of leptospirosis in Tanzania: A review of the current status, serogroup diversity and reservoirs

Leptospirainfection in people in the city of Durango, Mexico: a cross sectional study

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

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