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PubMed1980

Human glomerulonephritis and persistent non-polio enterovirus infection.

In a prospective work, we have studied the non-polio enterovirus (NPEV) excretion in spot urine and stools by four appropriate cell cultures, in four different groups: 20 exposed controls (GI), 88 patients with renal biopsy 'proven GN' (GII), 38 cases with 'proven nonGN' (GIII), and 9 with 'probable nonGN' (GIV). The positive excretion in stools and/or urine is respectively 0 and 5% in Group I, 14 and 33% in Group II, 6 and 19% in Group III, and 13 and 22% in Group IV. Viruria, the consequence of a viraemia, is therefore associated with 'proven GN' (p less than 0.05). In the majority of patients with positive NPEV excretion, we have made an additional but similar study, 3-12 months later. Persistent excretion was confirmed in 22/30 cases in Group II (73%) versus 0/8 in Group III-IV (p less than 0.001). These data concerned patients with membranoproliferative GN [5], membranous GN [6], mesangial IgA GN [6], endocapillary GN [2] or minimal lesions [3]. Thus we have demonstrated a significant relation between persistent NPEV urine/stools excretion and the occurrence of active GN in humans. Such persistent viral infections may represent the cause of some GN, probably mediated by an immune complex mechanism with viral antigens.

Enterovirus↗
PubMed2002

Development and evaluation of nucleic acid sequence based amplification (NASBA) for diagnosis of enterovirus infections using the NucliSens Basic Kit.

BACKGROUND: Molecular methods based on RNA amplification are needed for sensitive detection of enteroviruses in clinical samples. Many 'in house' methods based on reverse-transcribed PCR (RT-PCR) could be difficult to use in the routine diagnostic laboratory since they tend to be time-consuming, use reagents from many different suppliers and include non-routine procedures. OBJECTIVES: The aim of this study was to develop and evaluate methods based on nucleic acid sequence based amplification (NASBA) for detection of enterovirus sequences. STUDY DESIGN: 'In house' prepared and commercially available reagents were utilised to develop enterovirus-specific NASBA assays. Optimised methods were evaluated using clinical samples (cerebrospinal fluid, respiratory and stool samples), titred virus controls and in vitro produced synthetic RNA. Results for NASBA were compared with RT-PCR and virus culture. RESULTS: Kit-based reagents gave an equivalent sensitivity to the more laborious 'in house' molecular assays (NASBA and RT-PCR) on clinical material and controls. All molecular methods picked up enterovirus positive clinical samples that were not identified by culture. End point detection sensitivity for the NASBA assay based on the NucliSens Basic Kit was <or=1 tissue culture infective dose 50% of a range of enteroviruses or <100 copies RNA input. The assay was specific for enteroviruses and did not pick up high titre rhinovirus preparations. Enterovirus Basic Kit NASBA results for clinical samples were easily obtained within a single working day. CONCLUSIONS: NASBA is a suitable alternative to RT-PCR for sensitive amplification and detection of enterovirus sequences in a range of clinical specimens. The use of kit-based reagents will enable a wide range of laboratories to undertake molecular-based diagnostic procedures for RNA viruses and provide results within a time frame relevant to patient management.

Enterovirus↗
PubMed2003

Enterovirus infection and activation of human umbilical vein endothelial cells.

Gastrointestinal tract associated lymphoid tissue is considered to be the main replication site for enteroviruses. In order to invade tissues to reach pancreatic islets, cardiac muscles, and other secondary replication sites, the virus has to survive circulation in the blood and find a way to get through endothelial cells. In the present study, the susceptibility of human endothelial cells to infections caused by human parechovirus 1 and several prototype strains of enteroviruses, representing different species (human poliovirus, human enterovirus B and C), and acting through different receptor families was examined. Primary endothelial cells isolated from human umbilical vein by collagenase perfusion and also an established human endothelial cell line, HUVEC, were used. Primary endothelial cells were highly susceptible to several serotypes of enteroviruses (coxsackievirus A13, echoviruses 6, 7, 11, 30, and poliovirus 1). However, coxsackievirus A 9 and echovirus 1 infected only a few individual cells while human parechovirus 1 and coxsackie B viruses did not show evidence of replication in primary endothelial cells. In general, primary endothelial cells were more sensitive to infection-induced cytolytic effect than HUVEC. Activation of endothelial cells by interleukin-1beta did not change the pattern of enterovirus infection. Immunofluorescence stainings of infected primary endothelial cells showed that expression of activation markers, E-selectin, and intercellular adhesion molecule-1, was clearly increased by several virus infections and the former molecule also by exposing cells to UV-light inactivated coxsackieviruses. In contrast, human leukocyte antigen-DR expression was not increased by virus infection.

Cell Line↗