PubMed Health⌕ Search

Biomedical subjects

D W Brown

Publications and source records attributed to D W Brown.

At least 73 records · Page 4Linked to original sources

The role of environmental contamination with small round structured viruses in a hospital outbreak investigated by reverse-transcriptase polymerase chain reaction assay.

In May 1994 an outbreak of vomiting and diarrhoea occurred in a 28-bed long-stay ward for the mentally infirm. The predominant symptoms were vomiting, diarrhoea, malaise and abdominal pain lasting for approximately 12 h in most cases. The attack rate was 62% (13/21) for patients and 46% (16/35) for staff members. Infection control measures were implemented (containment of infectious individuals, hand hygiene among staff and environmental decontamination) and the ward was closed to admissions. Affected staff were excluded from contact with patients and their food until asymptomatic for 72 h. The outbreak lasted for 17 days. Faecal samples from nine symptomatic persons were negative for bacterial enteric pathogens, Giardia, Cryptosporidium and group A rotavirus. Electron microscopy of 12 faecal samples and one sample of vomitus revealed small round structured virus (SRSV) particles in one faecal sample. A further 30 faecal samples and seven vomitus samples were tested by reverse transcription polymerase chain reaction (RT-PCR) for SRSV of which 12 (40%) and 1 (14%) were positive respectively. Twenty-eight throat swabs from symptomatic and asymptomatic patients were collected, three (9.5%) of which were positive for SRSV by RT-PCR. Thirty-six environmental swabs were collected on the affected ward, and 11 (30%) were positive by RT-PCR. Positive swabs were from lockers, curtains and commodes and confined to the immediate environment of symptomatic patients. The distribution of contamination supports the rationale of cohorting sick patients.

Cross Infection↗

Characterization of a new genotype of measles virus detected in China and England.

We report the co-circulation of a new lineage of measles virus (MV) and an Edmonston-like (Ed-like) genotype of MV in China during 1995-7. Sequence analysis of 25 strains was performed on a 282 nucleotides (nt) region of the nucleoprotein (N) gene, a 450-nt region of the haemagglutinin (H) gene and a 152-nt region of the matrix (M) gene by direct sequencing of RT-PCR amplicons obtained from clinical specimens. The entire H gene was sequenced from two strains. The results showed that 24/25 Chinese strains belonged to a new genogroup and were distinct from the vaccine strains used in China and the UK, and also from MV strains previously described in Europe, Africa and the USA. The remaining strain was Ed-like. Two strains of the new genotype (IV) and one of the Ed-like genotype were also detected in the UK in 1996.

Adolescent↗

Salivary diagnosis of rubella: a study of notified cases in the United Kingdom, 1991-4.

Rubella infections, notified by general practitioners on the basis of a clinical diagnosis, were investigated by testing blood and saliva samples for specific IgM. Overall 52 (29%) of 178 cases with appropriately timed blood specimens were confirmed as recent rubella by IgM serology. Only 2 (3%) of 74 cases in children under 5 years were confirmed compared to 50 (48%) of 104 cases in older children and adults. The confirmation rate was lower (6%) in those with documented vaccination history than in those without (42%). The specificity of saliva rubella IgM testing compared to testing corresponding blood samples was 99%. The overall sensitivity of saliva rubella IgM testing was 81%. This rose to 90% if results from inappropriately timed specimens and specimens taking more than 1 week to reach the laboratory were excluded. A corresponding saliva rubella IgG test was 98% sensitive and 100% specific. Of 126 rubella IgM negative cases, 25 (20%) were positive for parvovirus B19 IgM. This study confirmed that rubella surveillance based on clinical reports is not specific. It also demonstrated that saliva samples, if taken 7-42 days after onset of illness and transported rapidly to the laboratory, are a feasible alternative to blood samples for rubella surveillance.

Adolescent↗

Prevalence and correlates of herpes simplex virus type 2 infection: evaluation of behavioural risk factors.

OBJECTIVE: To examine the prevalence and correlates of infection with herpes simplex virus type 2 (HSV-2) among sexually transmitted disease (STD) clinic attenders, we studied the prevalence of antibodies to HSV-2 and their association with risk behaviour. METHODS: Data were collected in a cross-sectional study among STD clinic attenders in Amsterdam. Seropositivity for HSV-2 was determined in 1798 serum samples by means of a monoclonal antibody-blocking enzyme-linked immunoassay. RESULTS: The prevalence of HSV-2 antibodies was higher than expected: 32.3% in a population in which 3% had current genital herpes and 8% gave a history of genital herpes. Of those with HSV-2 antibodies, only 18% had a history of genital herpes. A strong independent association with the presence of HSV-2 antibodies was found for sexual behaviour, more specifically: homosexual orientation, increasing number of years of sexual activity, increasing number of lifetime partners, number of past gonococcal infections, having receptive anal and (or) vaginal contact. CONCLUSION: The presence of HSV-2 antibodies had a strong association with past sexual behaviour and, for both sexes, with receptive anal intercourse. HSV-2 antibodies may be used as a surrogate marker of sexual risk behaviour in comparing different populations over time.

Adult↗

A nested reverse transcriptase PCR assay for detection of small round-structured viruses in environmentally contaminated molluscan shellfish.

We describe the evaluation of a nested reverse transcriptase PCR (RT-PCR) procedure for the detection of small round-structured viruses (SRSVs) in molluscan shellfish and the application of this assay for the detection of SRSVs in commercially produced shellfish and in shellfish implicated in outbreaks of gastroenteritis. The range of virus strains detected and the sensitivity of detection were evaluated by using a representative panel of 21 well-characterized SRSV strains. The nested RT-PCR detected 15 of 21 SRSVs, demonstrating that the assay detects a broad range of SRSVs including strains from both genogroup I and genogroup II. Seeding experiments showed the nested RT-PCR assay to be 10 to 1,000 times more sensitive than the single-round RT-PCR assay for the detection of SRSV in shellfish. SRSV-contaminated samples were identified by nested RT-PCR for shellfish grown in polluted harvesting areas and for shellfish associated with outbreaks of gastroenteritis which were negative by a previously described single-round RT-PCR. The assay was shown to be effective for investigation of virus elimination during commercial shellfish processing procedures such as depuration and relaying and has potential applications for monitoring at-risk shellfish harvesting areas, for investigation of SRSV contamination in shellfish from producers linked to gastroenteritis outbreaks, and for the direct detection of virus in shellfish implicated in outbreaks.

Polymerase Chain Reaction↗

A novel simian immunodeficiency virus (SIVdrl) pol sequence from the drill monkey, Mandrillus leucophaeus.

The drill monkey has been shown by serology and PCR to harbor a unique simian immunodeficiency virus (SIVdrl). A pol sequence, amplified from uncultured peripheral blood cells, is most closely related to the equivalent SIV sequences from the red-capped mangabey (SIVrcm), the sabaeus African green monkey (SIVagmSAB), and the chimpanzee (SIVcpz) and to the human immunodeficiency virus type 1 (HIV-1) sequence of humans. It is as yet unclear whether SIVdrl has a mosaic genome like SIVrcm and SIVagmSAB, is a member of the SIVcpz/HIV-1 lineage, or represents a novel primate lentivirus lineage.

Amino Acid Sequence↗

Epidemiology of Epstein-Barr virus infection in pre-adolescent children: application of a new salivary method in Edinburgh, Scotland.

STUDY OBJECTIVE: To describe the epidemiology of Epstein-Barr virus (EBV) among primary school children by testing saliva with a new EBV capsid antigen "G" antibody capture radioimmunoassay (GACRIA). DESIGN: A population based sample of 7 year old schoolchildren were followed up at age 11. SETTING: 30 randomly chosen primary schools in Edinburgh, Scotland. PARTICIPANTS: 552 schoolchildren. MEASUREMENTS: Data on risk factors for infection were collected by questionnaire at ages 7 and 11. Saliva samples collected at age 11 were examined by GACRIA for evidence of previous infection with EBV. For 102 subjects, a second salivary specimen collected approximately one month after the first sample was available for testing as a measure of the repeatability of the method. MAIN RESULTS: Unequivocal results were found in 91% of samples and the repeatability of the test was good (kappa = 0.71). Fifty six per cent of children had antibodies to EBV. In a logistic regression analysis, independent risk factors for infection were sharing a room (odds ratio 1.78, 1.14, 2.79), head of household's social class IV/V compared with I (odds ratio 2.87, 1.08, 7.34), and schools serving predominantly rented housing estates (odds ratio 2.3, 1.09, 4.84). CONCLUSION: This study is the first application of EBV viral capsid GACRIA to salivary samples. The method was successfully used to describe the epidemiology of EBV. In this study, characteristics of the home seemed to be more important than those of the school in determining the likelihood of infection with EBV.

Adult↗

Role of type specific herpes simplex virus serology in the diagnosis and management of genital herpes.

OBJECTIVES: To investigate the indications for the use of a type specific antibody test for herpes simplex virus in a department of genitourinary medicine in the United Kingdom. METHOD: Retrospective analysis of case records of 127 patients who accepted the test during a 20 month period. RESULTS/CONCLUSION: The test contributed to patient management in 79% of patients with recurrent genital ulceration of unknown cause. It was also useful for counselling a number of patients with initial episodes of disease and the asymptomatic partners of some patients when the partners were shown to possess antibodies specific to herpes simplex virus type 2. When evaluating sexual partners, the test was difficult to interpret if an isolate from the index case had not been typed. Access to viral typing may therefore be a greater priority than serological testing. As adverse psychological sequelae may follow the identification of an asymptomatic chronic infection, guidelines for the use of a type specific serological test are proposed.

Antibodies, Viral↗

Homotypic and heterotypic IgG and IgM antibody responses in adults infected with small round structured viruses.

Antibody responses to recombinant Norwalk (rNV) and Mexico (rMXV) viral capsid proteins were studied in 39 adults involved in outbreaks of gastroenteritis associated with genogroup 2 small round structured viruses (SRSVs). Nineteen individuals were involved in outbreaks associated with MXV-like strains and 20 in outbreaks associated with four other genogroup 2 SRSVs. IgG antibodies were measured in acute and convalescent sera using indirect enzyme-linked immunosorbent assay (ELISA), and IgM was measured by indirect and capture ELISAs. Nineteen (49%) patients demonstrated a significant rise in IgG to rMXV with four (10%) patients also showing anamnestic responses to rNV. Fourteen patients were positive in the rMXV IgM-capture ELISA, representing 74% of patients demonstrating IgG rises. IgG and IgM responses to rMXV were observed in both groups, although higher levels of responses were seen in adults infected with MXV-like strains than those infected with non-MXV genogroup 2 viruses. No significant IgM responses were observed to rNV. These results indicate that, following SRSV infection, adults show a rise in antibody which is broadly reactive to viruses within but not between genogroups, although greater homotypic than heterotypic responses are produced. These findings have implications for interpretation of seroepidemiological studies and serodiagnosis of SRSV infections using recombinant capsids.

Adult↗

A comparison of PCR with virus isolation and direct antigen detection for diagnosis and typing of genital herpes.

Patients attending the genitourinary medicine clinic at Watford General Hospital, UK, were examined for clinical signs of genital herpes infection. Genital swabs were taken from 194 patients (126 female, 68 male) who presented with genital ulceration or symptoms which were suggestive of genital herpes infection. Swabs from these patients were tested by three methods: (i) Detection of herpes simplex virus (HSV) antigen by direct HSV enzyme immunoassay (EIA), (ii) HSV isolation in Vero cell culture and (iii) HSV polymerase chain reaction (PCR). HSV was detected in 76 patients (39%) by EIA, in 93 (48%) by isolation in cell culture, and in 115 (59%) by PCR. Isolation by cell culture has been considered as the "gold standard" for the detection of HSV in genital lesions, but in this study HSV PCR was significantly more sensitive. Comparison of the three methods was as follows: Cell culture vs. PCR: Sensitivity 93/115 (80.9%), Specificity 79/79 (100%). HSV EIA vs. PCR: Sensitivity 75/115 (65.2%), Specificity 78/79 (98.7%). HSV EIA vs. Cell culture: Sensitivity 75/93 (80.7%), Specificity 100/101 (99%). EIA was less effective in detecting HSV among recurrent than among first episode infections, in comparison to culture or HSV PCR. This is the first comparison of HSV PCR with two other routine diagnostic methods for confirming genital herpes infection in a symptomatic population. The infecting HSV type was identified by restriction digestion of 108 HSV amplicons: HSV-1:37/108 (34%), HSV-2:71/108 (66%). In this population HSV-1 causes a significant proportion of genital herpes cases, and HSV-1 genital infection was detected in significantly more first episode infections (40.3%) than among recurrent infections (22.2%).

Animals↗

Adrenomedullin has mitogenic effects on human oral keratinocytes: involvement of cyclic AMP.

The effects of the novel vasoactive regulatory peptide, adrenomedullin, on human oral keratinocytes was investigated. Adrenomedullin, acting via its specific receptor, stimulated a dose-dependent increase in DNA synthesis, and, in addition, stimulated further changes in the cell cycle resulting in the proliferation of keratinocytes. When cells were incubated in the presence of increasing concentrations of adrenomedullin, there was a rapid and dose-dependent rise in intracellular cyclic AMP levels. Stimulation of mitogenesis and cell proliferation in these cells were mimicked by the cell permeable cAMP analogue, dibutyryl cAMP. Adrenomedullin-stimulated mitogenesis was attenuated by the adenylyl cyclase inhibitor SQ22,536, but was unaffected by inhibitors of PKC, tyrosine kinase or the CGRP receptor antagonist, CGRP(8-37). These data identify adrenomedullin as a new mitogenic regulatory peptide of keratinocytes acting via the cAMP cascade.

Adrenomedullin↗

Detection of IgG antibody to Epstein-Barr virus viral capsid antigen in saliva by antibody capture radioimmunoassay.

A 'G' antibody capture radioimmunoassay (GACRIA) to detect IgG to Epstein-Barr virus (EBV) viral capsid antigen (VCA) in saliva is described. The monoclonal antibody to EBV VCA used in the GACRIA bound non-specifically when testing saliva samples having a total IgG content of less than 2.0 mg/l, so giving false positive results. This problem was overcome by including 0.5% EBV-negative human serum in the monoclonal antibody diluent. The performance of the assay was then evaluated by comparing the GACRIA test on serum and saliva samples to indirect immunofluorescence assay (IFA) results on sera using a panel of paired serum/saliva samples. Compared to the corresponding serum IFA the saliva GACRIA had a sensitivity and specificity of 93.5 and 100%, respectively. Although less sensitive than IFA on serum samples, the saliva GACRIA is sufficiently sensitive to be used for epidemiological screening and will enable testing for anti-EBV VCA to be carried out easily and on a wide scale.

Antibodies, Viral↗

Epidemiology of Mexico virus, a small round-structured virus in Yorkshire, United Kingdom, between January 1992 and March 1995.

The epidemiology of the small round-structured virus, Mexico virus (MxV), was investigated in North and West Yorkshire, United Kingdom, between January 1992 and March 1995 using a type-specific antigen ELISA. The results indicate that an epidemic of MxV occurred during the winter of 1993-1994, when this strain was associated with 45 of 99 outbreaks and sporadic childhood cases of gastroenteritis. Only 4 MxV-like isolates were found during the 1992-1993 winter season and none in the 1994-1995 season. This descriptive epidemiologic study suggests that MxV has an epidemic pattern of infection.

Caliciviridae↗

Polio eradication: surveillance implications for the United Kingdom.

The requirements for certification of elimination of wild virus poliomyelitis will pose particular problems for some industrialized countries, such as the United Kingdom, where there has been no case detected for at least a decade. Systems of surveillance of poliomyelitis have been reviewed and potential weaknesses identified. When oral polio vaccine is routinely used, the rate of vaccine-associated cases provides an indication of the likelihood that if they occurred, wild virus cases would be detected. Acute flaccid paralysis surveillance was done for 3 years, but rates were lower than reported elsewhere and were accepted for certification purposes. Alternative techniques, such as surveillance of polioviruses, either in clinical samples or from the environment, may be developed in such countries. The ability to identify enteroviruses and to distinguish between wild and vaccine strains of polioviruses will give assurance that silent transmission of wild viruses is unlikely.

Disease Notification↗

The diversity of measles virus in the United Kingdom, 1992-1995.

Three distinct genotypes were identified amongst 50 measles virus (MV) strains characterized in the UK between 1992 and 1995 by direct sequencing of the RT-PCR products amplified from clinical specimens. All three genotypes were related to viruses previously reported in the United States or in continental Europe. Phylogenetic analyses of 255 and 152 nucleotide sequences from the N and M genes, respectively, generated very similar lineages. The degree of divergence between genotypes was 3.5-16% and 2.6-7.9% in the N and the M genes, respectively. MV genotypes which circulated during the 1970s and 1980s in the UK were not detected in this period. Comparison of the C-terminal regions of the N gene sequences of UK strains isolated or detected between 1974 and 1995 suggests that there have been multiple genotypes of MV circulating in the UK over the past 20 years.

Base Sequence↗

Mixed genogroup SRSV infections among a party of canoeists exposed to contaminated recreational water.

Samples of faeces collected from a party of canoeists involved in a gastroenteritis outbreak were examined by electron microscopy and RT-PCR for evidence of infection with SRSVs. A broadly reactive primer pair was used to detect SRSVs followed by application of genogroup-specific primers to SRSV-positive specimens. Exposure data were collected by means of a questionnaire. SRSVs were detected in 1/4 specimens examined by EM and 3/4 by RT-PCR. Genogrouping, and sequencing of PCR products revealed two distinct strains: a genogroup I strain, related to the Desert Shield virus, and a genogroup II strain, related to the Lordsdale virus to be associated with the outbreak. Exposure data indicated that capsising and eating food before getting changed were associated with an increased risk of gastroenteritis and was consistent with infection following the consumption of contaminated water. This study confirms the greater sensitivity of RT-PCR for the diagnosis of SRSV infections and its utility, when incorporating genogroup-specific primers, in establishing more complex epidemiological data.

Adolescent↗