Sadeghi Mohammadreza, Wang Yilin, Ramqvist Torbjörn, Aaltonen Leena-Maija, Pyöriä Lari, Toppinen Mari, Söderlund-Venermo Maria, Hedman Klaus
Virology, University of Helsinki, Helsinki, Finland.
Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
BMC Infect Dis. 2017 Jun 8;17(1):409. doi: 10.1186/s12879-017-2479-5.
In the past few years, eleven new human viruses have joined the two previously known members JCPyV and BKPyV of the Polyomaviridae family, by virtue of molecular methods. Serology data suggest that infections with human polyomaviruses (HPyVs) occur since childhood and the viruses are widespread in the general population. However, the viral persistence sites and transmission routes are by and large unknown. Our previous studies demonstrated that the four new HPyVs - KIPyV, WUPyV, MCPyV and TSPyV - were present in the tonsils, and suggested lymphoid tissue as a persistent site of these emerging human viruses. We developed a Luminex-based multiplex assay for simultaneous detection of all 13 HPyVs known, and explored their occurrence in tonsillar tissues of children and adults mostly with tonsillitis or tonsillar hypertrophy.
We set up and validated a new Luminex-based multiplex assay by using primer pairs and probes targeting the respective HPyV viral protein 1 (VP1) genes. With this assay we tested 78 tonsillar tissues for DNAs of 13 HPyVs.
The multiplex assay allowed for simultaneous detection of 13 HPyVs with high analytical sensitivity and specificity, with detection limits of 10-10 copies per microliter, and identified correctly all 13 target sequences with no cross reactions. HPyV DNA altogether was found in 14 (17.9%) of 78 tonsils. The most prevalent HPyVs were HPyV6 (7.7%), TSPyV (3.8%) and WUPyV (3.8%). Mixed infection of two HPyVs occurred in one sample.
The Luminex-based HPyV multiplex assay appears highly suitable for clinical diagnostic purposes and large-scale epidemiological studies. Additional evidence was acquired that the lymphoid system plays a role in HPyV infection and persistence. Thereby, shedding from this site during reactivation might take part in transmission of the newly found HPyVs.
在过去几年中，借助分子方法，11种新的人类病毒加入了多瘤病毒科先前已知的两个成员——JC多瘤病毒（JCPyV）和BK多瘤病毒（BKPyV）。血清学数据表明，人类多瘤病毒（HPyV）感染自儿童期就已发生，且这些病毒在普通人群中广泛传播。然而，病毒的持续存在部位和传播途径基本上尚不清楚。我们之前的研究表明，4种新的HPyV——KIPyV、WUPyV、MCPyV和TSPyV——存在于扁桃体中，并提示淋巴组织是这些新出现的人类病毒的一个持续存在部位。我们开发了一种基于Luminex的多重检测方法，用于同时检测所有已知的13种HPyV，并探究它们在主要患有扁桃体炎或扁桃体肥大的儿童和成人扁桃体组织中的存在情况。
我们使用靶向各HPyV病毒蛋白1（VP1）基因的引物对和探针，建立并验证了一种基于Luminex的新多重检测方法。利用该检测方法，我们检测了78份扁桃体组织中13种HPyV的DNA。
该多重检测方法能够以高分析灵敏度和特异性同时检测13种HPyV，检测限为每微升10⁻¹⁰拷贝，且能正确识别所有13个靶序列，无交叉反应。在78份扁桃体样本中，共14份（17.9%）检测到HPyV DNA。最常见的HPyV是HPyV6（7.7%）、TSPyV（3.8%）和WUPyV（3.8%）。1份样本中出现了两种HPyV的混合感染。
基于Luminex的HPyV多重检测方法似乎非常适合临床诊断目的和大规模流行病学研究。我们获得了更多证据，表明淋巴系统在HPyV感染和持续存在中发挥作用。因此，在病毒重新激活期间从该部位排出病毒可能参与了新发现的HPyV的传播。
