日本血吸虫感染的血清学诊断：一种蛋白质标记物的全基因组鉴定及其在中国现场研究中的诊断有效性评估。

Serodiagnosis of Schistosoma japonicum infection: genome-wide identification of a protein marker, and assessment of its diagnostic validity in a field study in China.

机构信息

Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.

Jiangxi Provincial Institute of Parasitic Diseases, Nanchang 330046, Jiangxi, China.

出版信息

Lancet Infect Dis. 2014 Jun;14(6):489-97. doi: 10.1016/S1473-3099(14)70067-2. Epub 2014 Mar 19.

DOI:10.1016/S1473-3099(14)70067-2

PMID:24656567

Abstract

BACKGROUND

Schistosomiasis remains a highly prevalent and serious parasitic disease. A major factor preventing its effective management is the scarcity of effective diagnostic tools. We did a genome-wide identification of diagnostic protein markers for schistosome infection and assessed their diagnostic validity in a field study.

METHODS

We predicted putative secreted proteins of Schistosoma japonicum (SjSPs) and expressed them as glutathione S-transferase (GST)-fusion proteins. The fusion proteins were arrayed on glutathione (GSH)-immobilised microplates and screened with serum samples from patients with schistosomiasis diagnosed by the Kato-Katz method. We further assessed an identified protein marker for sensitivity and specificity, first in infected serum samples collected from Jiangxi and Hunan Provinces, China, and then through a field study, done in two villages located in a high schistosomiasis-endemic area of the southeast of China.

FINDINGS

Of 204 recombinant proteins, 35 yielded seropositive reactions, eight showed strong immunoreactivity, and only one (SjSP-13) reacted to the entire panel of 14 archived samples. The reactivity of SjSP-13 to 476 serum samples showed 90·4% (95% CI 86·5-93·5) sensitivity and 98·9% (95% CI 95·9-99·9) specificity. Of 1371 residents enrolled in a field study from Dec 6, 2010, to June 23, 2011, only 74 individuals were identified as being egg-positive, whereas 465 were diagnosed as positive by the SjSP-13-based ELISA kit (rSP13-ELISA). Of the 394 individuals found egg-negative but rSP13-ELISA-positive, 363 (92·4%) were confirmed to be positive for schistosome infection by PCR detection of S japonicum SjR2 retrotransposon.

INTERPRETATION

The application of this sensitive, specific, and affordable rSP13-ELISA method should help reduce schistosomiasis transmission through targeted treatment of individuals, particularly with low intensity infections, and therefore support schistosomiasis control and elimination strategies.

FUNDING

National 973 project in China.

摘要

背景

血吸虫病仍然是一种高度流行和严重的寄生虫病。其有效管理的一个主要障碍是缺乏有效的诊断工具。我们对日本血吸虫（Schistosoma japonicum）的诊断蛋白标记物进行了全基因组鉴定，并在现场研究中评估了它们的诊断有效性。

方法

我们预测了日本血吸虫（Schistosoma japonicum）的假定分泌蛋白（SjSPs），并将其表达为谷胱甘肽 S-转移酶（GST）融合蛋白。融合蛋白被排列在谷胱甘肽（GSH）固定化微板上，并与通过加藤法诊断为血吸虫病的患者的血清样本进行筛选。我们进一步评估了一种鉴定的蛋白标记物的敏感性和特异性，首先是在中国江西和湖南两省采集的感染血清样本，然后是在中国东南部高血吸虫病流行地区的两个村庄进行的现场研究。

发现

在 204 个重组蛋白中，有 35 个产生了血清阳性反应，8 个显示出强烈的免疫反应，只有 1 个（SjSP-13）反应了整个 14 个存档样本。SjSP-13 对 476 个血清样本的反应显示出 90.4%（95%CI 86.5-93.5）的敏感性和 98.9%（95%CI 95.9-99.9）的特异性。在 2010 年 12 月 6 日至 2011 年 6 月 23 日期间，对参加现场研究的 1371 名居民进行了调查，只有 74 人被确认为卵阳性，而 465 人被 SjSP-13 基于 ELISA 试剂盒（rSP13-ELISA）诊断为阳性。在 394 名被发现卵阴性但 rSP13-ELISA 阳性的个体中，363 名（92.4%）通过 PCR 检测日本血吸虫 SjR2 反转录转座子被确认为日本血吸虫感染阳性。