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Rapid, actionable diagnosis of urban epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay
PLOS Neglected Tropical Diseases ( IF 3.8 ) Pub Date : 2017-09-15 , DOI: 10.1371/journal.pntd.0005940
Irina N. Riediger , Robyn A. Stoddard , Guilherme S. Ribeiro , Sueli M. Nakatani , Suzana D. R. Moreira , Irene Skraba , Alexander W. Biondo , Mitermayer G. Reis , Alex R. Hoffmaster , Joseph M. Vinetz , Albert I. Ko , Elsio A. Wunder

Background

With a conservatively estimated 1 million cases of leptospirosis worldwide and a 5–10% fatality rate, the rapid diagnosis of leptospirosis leading to effective clinical and public health decision making is of high importance, and yet remains a challenge.

Methodology

Based on parallel, population-based studies in two leptospirosis-endemic regions in Brazil, a real-time PCR assay which detects lipL32, a gene specifically present in pathogenic Leptospira, was assessed for the diagnostic effectiveness and accuracy. Patients identified by active hospital-based surveillance in Salvador and Curitiba during large urban leptospirosis epidemics were tested. Real-time PCR reactions were performed with DNA-extracted samples obtained from 127 confirmed and 23 unconfirmed cases suspected of leptospirosis, 122 patients with an acute febrile illness other than leptospirosis, and 60 healthy blood donors.

Principal findings

The PCR assay had a limit of detection of 280 Leptospira genomic equivalents/mL. Sensitivity for confirmed cases was 61% for whole blood and 29% for serum samples. Sensitivity was higher (86%) for samples collected within the first 6 days after onset of illness compared to those collected after 7 days (34%). The real-time PCR assay was able to detect leptospiral DNA in blood from 56% of serological non-confirmed cases. The overall specificity of the assay was 99%.

Conclusions

These findings indicate that real-time PCR may be a reliable tool for early diagnosis of leptospirosis, which is decisive for clinical management of severe and life-threatening cases and for public health decision making.



中文翻译:

使用基于致病性钩端螺旋体lipL32的实时PCR测定法快速,可行地诊断城市流行性钩端螺旋体病

背景

据保守估计,全球范围内有100万例钩端螺旋体病病例,死亡率为5-10%,对钩端螺旋体病进行快速诊断以做出有效的临床和公共卫生决策非常重要,但仍然是一个挑战。

方法

根据在巴西两个钩端螺旋体病流行地区进行的基于人群的平行研究,评估了检测病原钩端螺旋体中特有的基因lipL32的实时PCR检测方法的诊断效果和准确性。在大型城市钩端螺旋体病流行期间,对在萨尔瓦多和库里提巴进行的基于医院的主动监测发现的患者进行了测试。使用从127例确诊和23例疑似钩端螺旋体病病例,122例除钩端螺旋体病以外的急性发热性疾病患者和60名健康献血者中提取的DNA提取样品进行实时PCR反应。

主要发现

PCR检测的检出限为280个钩端螺旋体基因组当量/ mL。确诊病例的敏感性为全血61%,血清样本29%。与发病后7天(34%)相比,发病后6天之内的样品灵敏度更高(86%)。实时PCR检测能够从56%的血清学未确诊病例中检测出血液中的钩端螺旋体DNA。该测定法的总特异性为99%。

结论

这些发现表明,实时PCR可能是钩端螺旋体病早期诊断的可靠工具,对严重和威胁生命的病例的临床管理以及公共卫生决策具有决定性作用。

更新日期:2017-09-15
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