Revealing enterovirus infection in chronic human disorders: An integrated diagnostic approach.
Revealing enterovirus infection in chronic human disorders: An integrated diagnostic approach.
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DOI：
10.1038/s41598-017-04993-y
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发表时间：
2017-07-10
影响因子：
4.6
通讯作者：
Toniolo A
中科院分区：
文献类型：
--
作者：
Genoni A;Canducci F;Rossi A;Broccolo F;Chumakov K;Bono G;Salerno-Uriarte J;Salvatoni A;Pugliese A;Toniolo A
肠道病毒（EV）引起持续感染的特点是最小的复制和遗传变化。这些药物的分型可以补充疾病评估并阐明发病机制。在这里，我们报告了一个综合的方法，在人类样本中的EV检测是基于在细胞培养中的病毒的预富集，然后搜索病毒基因组和病毒抗原。对脊髓灰质炎后综合征、1型糖尿病和慢性心肌病的病例进行了调查。由于基于组织的方法需要侵入性程序，因此主要从血液中的病毒收集信息。采用针对所有EV类型的保守基因组区域（5′UTR，2C，3Dpol）的分子检测。与血浆或白细胞直接检测相比，在分子和免疫学检测之前将白细胞与细胞系共培养可显著提高EV的检出率。RT-PCR和测序的结果通过用一组EV特异性抗体染色细胞培养物来确认。序列和系统发育分析表明，C类（脊髓灰质炎病毒）的EV与脊髓灰质炎后综合征有关，而B类成员被发现在1型糖尿病和心肌病。该程序可用于研究不同EV与各种慢性神经系统、内分泌和心脏疾病的可能关联。
Enteroviruses (EVs) causing persisting infection are characterized by minimal replication and genetic changes. Typing of these agents may complement disease assessment and shed light on pathogenesis. Here we report an integrated approach for EV detection in human samples that is based on pre-enrichment of virus in cell culture before search for the viral genome and viral antigens. Cases of post-polio syndrome, type 1 diabetes, and chronic cardiomyopathy were investigated. As tissue-based approaches require invasive procedures, information was mainly gleaned from virus in blood. Molecular assays targeting conserved genome regions of all EV types (5′UTR, 2 C, 3Dpol) were employed. As compared to direct assays of plasma or leukocytes, the EV detection rate was significantly enhanced by co-culture of leukocytes with cell lines prior to molecular and immunologic tests. Results of RT-PCR and sequencing were confirmed by staining cell cultures with a panel of EV-specific antibodies. Sequence and phylogenetic analysis showed that EVs of the C species (polioviruses) were associated with the post-polio syndrome, while members of the B species were found in type 1 diabetes and cardiomyopathy. The procedure may be used for investigating the possible association of different EVs with a variety of chronic neurologic, endocrine, and cardiac disorders.
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病毒学的单细胞基因组学。
DOI：
10.3390/v8050123
发表时间：
2016-05-04
期刊：
Viruses
影响因子：
--
作者：
Ciuffi A;Rato S;Telenti A
通讯作者：
Telenti A
影响因子：
6.4
作者：
Li, Li;Ivanova, Olga;Sutter, Roland W.
通讯作者：
Sutter, Roland W.
影响因子：
9.9
作者：
Cavalcante, P.;Barberis, M.;Bernasconi, P.
通讯作者：
Bernasconi, P.
影响因子：
12.7
作者：
Hammerstad, Sara Salehi;Tauriainen, Sisko;Dahl-Jorgensen, Knut
通讯作者：
Dahl-Jorgensen, Knut
影响因子：
7.7
作者：
Lundberg, Marcus;Krogvold, Lars;Skog, Oskar
通讯作者：
Skog, Oskar