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采用多重 qPCR 对巴贝西虫和伯氏疏螺旋体感染的有症状和无症状患者样本进行高效检测。

Efficient detection of symptomatic and asymptomatic patient samples for Babesia microti and Borrelia burgdorferi infection by multiplex qPCR.

机构信息

Department of Microbiology, Biochemistry and molecular Genetics, Rutgers New Jersey Medical School, Newark, New Jersey, United States of America.

The Gedroic Center, Morristown, NJ, United States of America.

出版信息

PLoS One. 2018 May 10;13(5):e0196748. doi: 10.1371/journal.pone.0196748. eCollection 2018.

Abstract

BACKGROUND

Tick-borne infections have been increasing steadily over the years, with co-infections with Borrelia burgdorferi and Babesia microti/divergens emerging as a serious health problem. B. burgdorferi is a spirochetal bacterium that causes Lyme disease while protozoan pathogens belonging to Babesia species are responsible for babesiosis. Currently used serological tests do not always detect acute Lyme disease or babesiosis, and fail to differentiate cured patients from those who get re-infected. This is a major problem for proper diagnosis particularly in regions endemic for tick-borne diseases. Microscopy based evaluation of babesiosis is confirmatory but is labor intensive and insensitive such that many asymptomatic patients remain undetected and donate blood resulting in transfusion transmitted babesiosis.

RESULTS

We conducted multiplex qPCR for simultaneous diagnosis of active Lyme disease and babesiosis in 192 blood samples collected from a region endemic for both diseases. We document qPCR results obtained from testing of each sample three times to detect infection with each pathogen separately or together. Results for Lyme disease by qPCR were also compared with serological tests currently used for Lyme disease when available. Considering at least two out of three test results for consistency, 18.2% of patients tested positive for Lyme disease, 18.7% for co-infection with B. burgdorferi and B. microti and 6.3% showed only babesiosis.

CONCLUSIONS

With an 80% sensitivity for detection of Lyme disease, and ability to detect co-infection with B. microti, multiplex qPCR can be employed for diagnosis of these diseases to start appropriate treatment in a timely manner.

摘要

背景

近年来,蜱传感染稳步增加,伯氏疏螺旋体(Borrelia burgdorferi)和微小巴贝斯虫/分歧巴贝斯虫(Babesia microti/divergens)的合并感染成为一个严重的健康问题。伯氏疏螺旋体是一种螺旋体细菌,可引起莱姆病,而属于巴贝斯虫属的原生动物病原体则引起巴贝斯虫病。目前使用的血清学检测方法并不总是能检测到急性莱姆病或巴贝斯虫病,也无法区分已治愈的患者和再次感染的患者。这是一个重大问题,特别是在蜱传疾病流行的地区,对正确诊断至关重要。基于显微镜的巴贝斯虫病评估是确认性的,但劳动强度大且不敏感,以至于许多无症状患者未被发现并捐献血液,导致输血传播的巴贝斯虫病。

结果

我们对来自两种疾病流行地区的 192 份血液样本进行了多重 qPCR 检测,以同时诊断活动性莱姆病和巴贝斯虫病。我们记录了对每个样本进行三次测试的 qPCR 结果,以分别或共同检测每个病原体的感染情况。当有可用的血清学检测方法时,我们将 qPCR 检测莱姆病的结果与目前用于莱姆病的血清学检测方法进行了比较。考虑到至少有两个测试结果的一致性,18.2%的患者莱姆病检测呈阳性,18.7%的患者同时感染伯氏疏螺旋体和微小巴贝斯虫,6.3%的患者仅表现为巴贝斯虫病。

结论

多重 qPCR 检测对莱姆病的检测灵敏度为 80%,并且能够检测到微小巴贝斯虫的合并感染,可用于这些疾病的诊断,以便及时开始适当的治疗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec7d/5945202/e70ef3174ae4/pone.0196748.g001.jpg

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