Department of Clinical Medicine, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka.
Institute of Biochemistry, Molecular Biology and Biotechnology, University of Colombo, Colombo, Sri Lanka.
PLoS Negl Trop Dis. 2020 May 19;14(5):e0008309. doi: 10.1371/journal.pntd.0008309. eCollection 2020 May.
Leptospirosis is endemic in Sri Lanka. There is a need for updated seroprevalence studies in endemic areas, to improve the understanding of disease dynamics, risk factors, control methods, and for clinical diagnosis. The cut-off titres for the microscopic agglutination test (MAT) for diagnosis of acute leptospirosis depend on community seroprevalence, and can vary based on locality and serovar. This study aimed to identify the seroprevalence, geographical determinants, and associations of seropositivity of leptospirosis in the district of Colombo in Sri Lanka, and to determine diagnostic cut-off titres for MAT in the community studied. This study utilized a stratified cluster sampling model in the Colombo district of Sri Lanka, to sample individuals living in urban and semi-urban areas. Serovar specific MAT titres were measured on recruited individuals using a panel of saprophytic (Leptospira biflexa) and 11 pathogenic Leptospira spp. serovars. Associations between environmental risk factors and MAT positivity were examined, with location mapping using GIS software. A total of 810 individuals were included. The mean age was 51.71 years (SD 14.02) with male predominance (60%). A total of 429 (53%) tested positive at a titer of 1/40 or more for the saprophytic Leptospira biflexa serovar Patoc. Pathogenic serovar MAT was positive at a titer of 1/40 or more for at least one serovar in 269 (33.2%) individuals. From the perspective of screening for clinical disease, serovar-specific cut-off titres of 1/80 for Leptospira spp. serovars Hebdomadis, Icterohaemorrhagiae, Pomona, Ratnapura and Patoc, 1/160 for serovars Pyrogenes and Cynopteri, and 1/40 for other serovars were determined, based on the 75th quartile MAT titre for each serovar. Serovar Pyrogenes (15.9%) had the highest seroprevalence, with serovars Ratnapura, Bankinang and Australis accounting for 9.9%, 9.6% and 9.3% respectively. When the proposed new cut-offs were applied, Bankinang(9.6%) Australis(9.3%), Pyrogenes(6.9%) and Ratnapura(6.9%) were the most prevalent serovars. No significant differences in seroprevalence or serovar patterns were noted between urban and semi-urban settings. Individuals seropositive for Australis, Ratnapura and Icterohaemorrhagiae were clustered around main water bodies as well as around smaller tributaries and paddy fields. Those positive for the serovar Pyrogenes were clustered around inland tributaries, smaller water sources and paddy fields. Associations of MAT positivity included high risk occupational exposure, environmental exposure including exposure to floods, bathing in rivers and lakes, using well-water for bathing, contact with stagnant water, propensity to skin injuries, presence of rats in the vicinity, and proximity to water sources. For pathogenic serovars, high-risk occupational exposure remained statistically significant following adjustment for other factors (adjusted OR = 2.408, CI 1.711 to 3.388; p<0.0001; Nagelkerke R2 = 0.546). High risk occupational exposure was determined to be independently associated with seropositivity. Baseline community MAT titres vary according to serovar, and presumably the locality. Testing against saprophytic serovars is unreliable. Thus, diagnostic MAT titre cut-offs should be determined based on region and serovar, and the use of a single diagnostic MAT cut-off for all populations is likely to result in false negatives.
钩端螺旋体病在斯里兰卡流行。在流行地区,需要更新血清流行率研究,以提高对疾病动态、危险因素、控制方法的了解,并进行临床诊断。显微镜凝集试验(MAT)用于诊断急性钩端螺旋体病的截止滴度取决于社区血清流行率,并且可以根据地理位置和血清型而有所不同。本研究旨在确定斯里兰卡科伦坡地区钩端螺旋体病的血清流行率、地理决定因素和血清阳性率的相关性,并确定研究社区中 MAT 的诊断截止滴度。本研究利用分层聚类抽样模型在斯里兰卡科伦坡地区抽取居住在城市和半城市地区的个体。使用一组腐生性(双曲钩端螺旋体)和 11 种致病性钩端螺旋体血清型,对招募的个体进行血清型特异性 MAT 滴度测量。使用 GIS 软件对环境危险因素与 MAT 阳性之间的相关性进行了检查,并进行了位置映射。共纳入 810 人。平均年龄为 51.71 岁(SD 14.02),男性为主(60%)。共有 429 人(53%)在腐生性双曲钩端螺旋体血清型 Patoc 的滴度为 1/40 或更高时呈阳性。致病性血清型 MAT 在至少一种血清型中滴度为 1/40 或更高的个体有 269 人(33.2%)。从临床疾病筛查的角度来看,基于每个血清型的 75 分位 MAT 滴度,确定了针对钩端螺旋体 spp.血清型 Hebdomadis、Icterohaemorrhagiae、Pomona、Ratnapura 和 Patoc 的 1/80、血清型 Pyrogenes 和 Cynopteri 的 1/160、其他血清型的 1/40 的血清型特异性截断滴度。血清型 Pyrogenes(15.9%)的血清流行率最高,血清型 Ratnapura、Bankinang 和 Australis 分别占 9.9%、9.6%和 9.3%。当应用新提出的截止值时,Bankinang(9.6%)、Australis(9.3%)、Pyrogenes(6.9%)和 Ratnapura(6.9%)是最流行的血清型。城乡环境之间的血清流行率或血清型模式无显著差异。对 Australis、Ratnapura 和 Icterohaemorrhagiae 呈血清阳性的个体聚集在主要水体周围,以及较小的支流和稻田周围。对血清型 Pyrogenes 呈血清阳性的个体聚集在内陆支流、较小的水源和稻田周围。MAT 阳性的相关性包括高风险职业暴露、包括暴露于洪水、在河流和湖泊中洗澡、使用井水洗澡、接触死水、皮肤受伤倾向、附近有老鼠以及靠近水源等环境暴露。对于致病性血清型,在调整其他因素后(调整后的 OR = 2.408,CI 1.711 至 3.388;p<0.0001;Nagelkerke R2 = 0.546),高风险职业暴露仍然具有统计学意义。高风险职业暴露被确定为与血清阳性独立相关。基础社区 MAT 滴度根据血清型和地理位置而有所不同。针对腐生性血清型的检测不可靠。因此,诊断 MAT 截止值应根据地区和血清型确定,并且使用单一诊断 MAT 截止值对所有人群进行检测可能会导致假阴性。