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液滴数字 PCR 诊断人类血吸虫病:不同临床样本中寄生虫无细胞 DNA 的检测。

Droplet Digital PCR Diagnosis of Human Schistosomiasis: Parasite Cell-Free DNA Detection in Diverse Clinical Samples.

机构信息

Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia.

School of Public Health, University of Queensland, Brisbane, Australia.

出版信息

J Infect Dis. 2017 Dec 19;216(12):1611-1622. doi: 10.1093/infdis/jix521.

Abstract

BACKGROUND

Schistosomiasis japonica remains a major public health and socioeconomic concern in Southeast Asia. Sensitive and accurate diagnostics can play a pivotal role in achieving disease elimination goals.

METHODS

We previously reported a novel droplet digital polymerase chain reaction (ddPCR) assay targeting the mitochondrial gene nad1 to diagnose schistosomiasis japonica. The tool identified both prepatent and patent infections using Schistosoma japonicum DNA isolated from serum, urine, salivary glands, and feces in a murine model. The assay was validated here using clinical samples collected from 412 subjects resident in an area moderately endemic for schistosomiasis in the Philippines.

RESULTS

S. japonicum DNA present in human stool, serum, urine, and saliva was detected quantitatively with high sensitivity. The capability to diagnose cases of human schistosomiasis using noninvasively collected clinical samples, the higher level of sensitivity obtained compared with the microscopy-based Kato-Katz test, and the capacity to quantify infection intensity have important public health implications for schistosomiasis control and programs targeting other neglected tropical diseases.

CONCLUSIONS

This verified ddPCR method represents a valuable new tool for the diagnosis and surveillance of schistosomiasis, particularly in low-prevalence and low-intensity areas approaching elimination and in monitoring where disease emergence or re-emergence is a concern.

摘要

背景

日本血吸虫病仍然是东南亚地区一个主要的公共卫生和社会经济关注问题。敏感和准确的诊断可以在实现消除疾病目标方面发挥关键作用。

方法

我们之前报道了一种针对线粒体基因 nad1 的新型液滴数字聚合酶链反应(ddPCR)检测方法,用于诊断日本血吸虫病。该工具使用从血清、尿液、唾液腺和粪便中分离的日本血吸虫 DNA ,在小鼠模型中鉴定了未成熟和成熟感染。在此,使用从菲律宾一个日本血吸虫中度流行地区的 412 名居民采集的临床样本对该检测进行了验证。

结果

定量检测到人类粪便、血清、尿液和唾液中的日本血吸虫 DNA,具有很高的灵敏度。使用非侵入性采集的临床样本诊断人类血吸虫病的能力、与基于显微镜的加藤氏检测相比获得的更高灵敏度以及定量感染强度的能力,对血吸虫病控制和针对其他被忽视的热带病的方案具有重要的公共卫生意义。

结论

这种经过验证的 ddPCR 方法代表了一种用于诊断和监测血吸虫病的有价值的新工具,特别是在接近消除的低流行率和低感染强度地区,以及在监测疾病出现或再次出现的情况下。

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