The number and range of enteroviruses isolated in the Regional Virus Laboratory, Glasgow during 1977–1997 was determined. Over this period, 3,039 enterovirus isolations were reported. The echoviruses represented 67% of isolations with echovirus 4 (due to an outbreak in 1990), echovirus 30 and echovirus 11 being the most frequently isolated types. The pattern of prevalence of non-polio enteroviruses had changed from the previous 20-year period with echovirus types isolated more often (77% vs. 55.4%) and coxsackieviruses isolated less often (23% vs. 44.6%). The polymerase chain reaction (PCR) introduced into the routine diagnostic service in 1996 increased the detection of enteroviruses from cerebrospinal fluid samples compared with traditional cell culture methods. Finally, the 5′ nontranslated region (NTR, bases 63–475) and the VP4/VP2 region (bases 581–1199) of selected echovirus 30 and coxsackie B3 isolates were sequenced. These represented endemic and epidemic types respectively and were shown to be closely related within their type, but different from the published sequences. The current echovirus 30 strains differed from 1966 isolates by 16–20% in both the 5′ NTR and VP4/VP2 regions. The coxsackie B3 isolates, predominant in 1997 after 5 years of absence, were also dissimilar from previously isolated strains, causing a small outbreak. J. Med. Virol. 58:304–312, 1999. © 1999 Wiley-Liss, Inc.

Summary Aims This study was designed to assess further the possible links between enterovirus infection and Type 1 diabetes mellitus (DM). Methods Sera from 110 children in the age range 0–15 years was obtained shortly after the diagnosis of Type 1 DM, in paediatric centres throughout the UK. They were tested for the presence of enteroviral sequences by the polymerase chain reaction (PCR) of the 5′ nontranslated region (5′ NTR). One hundred and eighty‐two controls tested were matched for age, geographical location and time of year. Results A significantly greater number of diabetic children (27% vs. 4.9%, P <0.005) had evidence of enteroviral RNA sequences. Proportionally, more younger children were enterovirus PCR positive, thus eight out of 20 children aged ≤2 years were enterovirus PCR positive. Sequence analysis showed that there was considerable variation in the sequences detected, although all appeared to be of the coxsackie/echovirus type. Conclusion This study re‐emphasizes that a link exists between enteroviral infection and the onset of Type 1 DM, particularly at a very early age, and suggests that these viruses are aetiologically important in diabetes in a significant proportion of children.

This report describes classical Type 1 insulin deficient diabetes mellitus (DM) arising in twins aged 14 months, both of whom had evidence of enterovirus infection. The diagnosis of Type 1 DM was made in the second twin within 12 days of the first. Enterovirus infection was detected in each twin at diagnosis by polymerase chain reaction (PCR). Both twins were negative for enterovirus by PCR 5 months following diagnosis, although both were then positive for islet cell antibodies. Sequencing of the amplicons produced by PCR suggested that the viruses from each twin were not the same but that they were both variants related to echovirus 6. © 1998 John Wiley & Sons, Ltd.

We have sought evidence of enteroviral persistence in humans. Eight individuals with chronic fatigue syndrome (CFS) were positive for enteroviral sequences, detected by PCR in two serum samples taken at least 5 months apart. The nucleotide sequence of the 5' non-translated region (bases 174-423) was determined for each amplicon. Four individuals had virtually identical nucleotide sequences ( > 97%) in both samples. The sequence pairs also each had a unique shared pattern indicating that the virus had persisted. In one individual (HO), it was clear that there had been infection with two different enteroviruses. In the remaining three individuals, the lack of unique shared features suggested that re-infection had occurred, rather than persistence. With the exception of HO, the sequences fell into a subgroup that is related to the Coxsackie B-like viruses.

We used a nested polymerase chain reaction (nPCR) to seek evidence for enteroviruses in clinical samples from patients with symptoms of aseptic meningitis. When compared with conventional virus isolation methods on a total of 366 samples collected during 1994-1995, an increase in positivity from 6% to 27% was shown. The results indicate that nPCR would be a valuable aid to the laboratory diagnosis of enteroviral infections as it can detect those enteroviruses that cannot be identified by current isolation methods.

Coxsackie B enteroviruses have been implicated repeatedly as agents associated with chronic fatigue syndrome (CFS). The objective of this study was to compare the serological evidence for the presence of Coxsackie B virus neutralising antibody, with the polymerase chain reaction (PCR) detecting a portion of the 5' nontranslated region (NTR) of the enterovirus genome. Serum samples from 100 chronic fatigue patients and from 100 healthy comparison patients were used in this study. In the CFS study group, 42% patients were positive for enteroviral sequences by PCR, compared to only 9% of the comparison group. Using the neutralisation assay, 34% of study patients were positive, compared to 41% of comparison patients. In the study group, 66/100 patient results correlated, i.e., they were either positive/positive or negative/negative for both tests. Of those that did not correlate, the majority were PCR-positive/Coxsackie B antibody-negative (21/34). In the comparison group, 58/100 patient results correlated. Of those that did not, the majority were PCR-negative/Coxsackie B antibody-positive (37/42). The Coxsackie B antibody neutralisation assay was not able to differentiate the CFS study group from the healthy comparison group, and thus the clinical relevance of this assay may be questioned. The PCR assay did differentiate the two groups with significantly more CFS patients having evidence of enterovirus than the comparison group.

Infection with Coxsackie B viruses has been linked to insulin-dependent diabetes mellitus. Nine of 14 serum samples (64%) taken from children at the onset of diabetes were positive for enterovirus RNA by PCR. All of the children were under age six, and five were under age three. By contrast, enterovirus sequences were detected in only two of 45 serum samples from appropriate comparison children (4%). Sequences from six of the positive patients showed strong homology with Coxsackie B3 and B4 viruses, and there were some common patterns among the sequences from infected diabetic children. This is evidence for a role for enteroviruses in childhood diabetes.

This study used phylogenetic analysis based on a region of the 5' non-translated region (5'NTR) of a variety of enteroviral sequences to compare sequences associated with chronic fatigue syndrome (CFS) and those from enteroviruses causing acute infections. Direct sequencing of PCR products was used to obtain the nucleic acid sequences from CFS patients. The inferred phylogenetic tree identified three groupings, one correlating with the diagnosis of CFS. The analysis identified a close relationship between the chronic fatigue enteroviral sequences, and showed that 19/20 were distinct from previously described enteroviruses. These results suggest there is persistence of enterovirus infection in some CFS patients and indicate the presence of distinct novel enterovirus sequences.

Objective-To investigate the association enteroviruses with motor neurone disease, known as amyotrophic lateral sclerosis.Design-Analysis by enterovirus polymerase chain reaction of wax embedded material from spinal cords taken at necropsy from subjects with motor neurone disease and from age and sex matched controls.Setting-Specimens were collected in the west of Scotland and in London between 1982 and 1992.Results-Sequences specific for a non-poliovirus type enterovirus were detected in spinal cord tissue from subjects with motor neurone disease. Amplification of a 414 base RNA target sequence in the conserved enterovirus 5' untranslated region from wax embedded tissue sections was successful in tissue from eight of 11 cases of sporadic motor neurone disease, one of two cases of familial motor neurone disease, and the one case of poliomyelitis, but not in the six matched controls or one case of antecedent poliomyelitis. In addition, sequences were detected in spinal cords from one monkey infected with wild type poliovirus and one monkey infected with polio vaccine. Comparison of sequences from cases of motor neurone disease with sequences of corresponding regions of the 5' untranslated regions of known picornaviruses showed them to be tightly grouped within the enterovirus genus closely related to coxsackievirus type B but not to polioviruses. Sequences derived from different parts of the spinal cord of the same subjects were identical, but sequences differed between individual subjects.Conclusions-Conserved enteroviral sequences closely related to coxsackie B virus sequences were detectable in spinal cords from subjects with sporadic motor neurone disease and from one subject with possible familial motor neurone disease.

The detection of specific RNA species in wax-embedded tissue sections using the polymerase chain reaction (PCR) means that gene expression can be studied and RNA viruses detected in stored histological tissue samples. This technique potentially allows the distribution of gene expression and viral replication to be studied in finely subdivided tissues. A technique is presented that has been used successfully to detect short RNA target sequences (130-420 bases) from proto-oncogene Abelson, human enteroviruses, and the sheep retrovirus Maedi-Visna virus using RNA PCR in single wax sections (20-30 microns). Various tissues were used which had not been deliberately prepared for this purpose. In a simple procedure hot xylene dewaxing is followed by acid phenol extraction of RNA and RNA PCR.

Reviews in Medical VirologyVolume 2, Issue 1 p. 51-51 Book Review Aids research reviews, Volume 1. Edited by W. C. Koff, F. Wong-Staal and R. C. Kennedy, Marcell Dekker Inc., New York (1991) G. B. Clements, G. B. Clements Consultant Virologist, Institute of Virology, University of Glasgow, Glasgow, UkSearch for more papers by this author G. B. Clements, G. B. Clements Consultant Virologist, Institute of Virology, University of Glasgow, Glasgow, UkSearch for more papers by this author First published: March 1992 https://doi.org/10.1002/rmv.1980020109AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume2, Issue1March 1992Pages 51-51 RelatedInformation

We have endeavoured to find immunological indications of chronic virus infection in patients with chronic fatigue syndrome (myalgic encephalomyelitis) and to investigate immune responsiveness to viruses in such patients in comparison with normal subjects and patients with muscular dystrophy. Levels of circulating IgM immune complexes were elevated (above the 95% normal control range) in 10 (17%) of 58 patients with chronic fatigue syndrome, which was not significantly different from the normal controls or from dystrophy controls (by Mann Whitney U test). Levels of IgG complexes were only increased in 10% of patients. Lymphocyte proliferation in response to concanavalin A (Con A), assessed by increase in 3H-thymidine incorporation, did not differ between 14 patients and 18 normal subjects. The proliferative response to Coxsackie B virus antigen did not differ between chronic fatigue patients and normal subjects when expressed either as an increase in counts or as a stimulation index. Adjustment of the counts in relation to the proliferation response to Con A, as an indication of the overall proliferative response of the cell preparation, did not reveal any hidden difference. IgM antibodies to Coxsackie B viruses were not found in any of 20 patients and in 1 of 20 dystrophy controls. Significant levels of neutralizing antibodies to Coxsackie B viruses 1-5 were found in 6 out of 19 (32%) patients compared with 4 out of 17 (24%) dystrophy controls, which does not differ from currently expected normal incidence. Antibody titres to other respiratory viruses were also not notably different between the patient and control groups. In conclusion we can find no evidence for a definable viral aetiology for the chronic fatigue syndrome, neither in terms of a persistent infection nor an altered ability to respond to virus.

Mycoplasma pneumoniae is the second most common cause of community-acquired pneumonia. Infection is found worldwide and epidemics are said to occur in 4-yearly cycles. In Scotland this pattern has been noted since 1982 and, in common with England and Wales as well probably as other parts of Europe, there is a current epidemic which began in the autumn of 1990. The disease has been noted predominantly in children and young adults, with lower respiratory tract infection as the most common manifestation. At present, diagnosis is based on a serological response and various tests are available for detecting both primary infection and reinfection. In view of the present epidemic, initial treatment of respiratory-tract infection, especially in children and young adults, should include adequate cover against Mycoplasma pneumoniae.

Objective-To determine the presence of enteroviral sequences in muscle of patients with the postviral fatigue syndrome.Design-Detection of sequences with the polymerase chain reaction in a well defined group of patients with the syndrome and controls over the same period.Setting-Institute of Neurological Sciences, Glasgow.Subjects-60 consecutive patients admitted to the institute with the postviral fatigue syndrome who had undergone extensive investigation to exclude other conditions. 41 controls from the same catchment area without evidence of fatigue, all undergoing routine surgery.Main outcome measures-Routine investigations, serological screen for antibodies to a range of viruses, and presence of enteroviral RNA sequences in muscle biopsy specimens.Results-15 (25%) patients and 10 (24.4%) controls had important serological findings. 12 patients had neutralising antibody titres of greater-than-or-equal-to 256 to coxsackieviruses B1-5 (six positive for enteroviral RNA sequences, six negative); three were positive for Epstein-Barr virus specific IgM (two positive, one negative). Six controls had similar neutralising antibody titres to coxsackieviruses (all negative); one was positive for Epstein-Barr virus specific IgM (negative); and three had titres of complement fixing antibody greater-than-or-equal-to 256 to cytomegalovirus (all negative). Overall, significantly more patients than controls had enteroviral RNA sequences in muscle (32/60, 53% v 6/41, 15%; odds ratio 6.7, 95% confidence interval 2.4 to 18.2). This was not correlated with duration of disease, patient and age, or to raised titres of antibodies to coxsackieviruses B1-5.Conclusions-Persistent enteroviral infection of muscle may occur in some patients with postviral fatigue syndrome and may have an aetiological role.

The production of a 57K stress protein (StrP) after herpes simplex virus type 1 (HSV-1) infection was examined in cultured neonatal rat retinal cells. StrP expression in individual cells was identified using a monoclonal antibody, TI56. Indirect immunofluorescence of uninfected retinal cultures showed that approximately 40% of cells expressed neurofilament (NF+) and 5% expressed a low level of StrP. Following HSV infection the proportion of NF+ cells decreased while the proportion of StrP positive cells became greater and the intensity of staining increased. The number of cells labelled with a polyclonal anti-HSV antibody increased with time after infection. Retinal neurons in culture can be infected with HSV, after which StrP expression is significantly upregulated.