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W D Cubitt

Publications and source records attributed to W D Cubitt.

At least 37 records · Page 2Linked to original sources
PubMed1996

Seroepidemiology of HTLV-I in relation to that of HIV-1 in the Gauteng region, South Africa, using dried blood spots on filter papers.

The seroprevalence of human T-lymphotropic virus type I (HTLV-I), in relation to that of human immunodeficiency virus type I (HIV-1), was determined in a comparative unlinked anonymous antenatal and neonatal (for indirect measurement of maternal antibodies) serosurvey in the Gauteng region of South Africa, using dried blood spots (DBS) and modified particle agglutination assays. Samples were confirmed to be antibody positive by western blot. A total of 2582 DBS collected during 1993 and 1994 from subjects of African, European and coloured origin were tested. Ten were confirmed as positive for HTLV-I and 128 for HIV-1. No antibodies to HTLV-I or HIV-1 were demonstrated in the 221 women of European and coloured origin who were screened. The HTLV-I seroprevalence rate in subjects of African origin appeared to increase from 0% in 1993 to 0.49% in 1994, while HIV-1 seroprevalence rates of 5.2% and 5.4% were recorded for 1993 and 1994 respectively. No significant differences in HTLV-I and HIV-1 seroprevalence rates were noted for the two areas investigated. These results indicate that HTLV-I should be included in infection control programs, and provide baseline data for monitoring the possible spread of HTLV-I in the heterosexual population in this region.

Black People↗
PubMed1996

An epidemiological investigation of Norwalk virus infection in South Africa.

A study was carried out to determine the incidence and seroprevalence of Norwalk virus (NV) in the Pretoria area, South Africa, using a recombinant NV (rNV) immunoassay for the detection of serum IgG antibodies. Maternal antibody was detectable in infants' sera up to approximately 6 months of age. Infection with NV was detected serologically in the second year of life and the seroprevalence of NV IgG rose from 37.1% at 7-11 months of age to 62.1% by the age of 40 years. No significant differences in seroprevalence of NV IgG antibody was evident between subjects of European or African ethnic origin, where overall seroprevalence rates were 56.4% and 53.9% respectively.

Adolescent↗
PubMed1995

Enzyme immunoassay using baculovirus-expressed human calicivirus (Mexico) for the measurement of IgG responses and determining its seroprevalence in London, UK.

The use of an enzyme immunoassay (EIA) employing a baculovirus-expressed recombinant human calicivirus (Mexico virus, MxV) for the detection of IgG-specific antibodies is described. MxV appeared to be related antigenically to a strain of small round structured virus, SRSV/UK4/Leeds/91, which had previously been shown by solid phase immune electron microscopy (SPIEM) to be related to Snow Mountain agent (SMA). One other outbreak which occurred in San Anita, USA in 1980 and was due to consumption of contaminated water was caused by a virus antigenically related to MxV. Volunteers and patients who developed significant IgG responses to rMxV showed anamnestic IgG responses (2 to 4-fold) in the recombinant Norwalk virus (rNV) IgG assay. Patients and volunteers who were known to have been infected with several other strains of calicivirus/small round spherical viruses (SRSV) including NV and SRSV UK3 showed no significant antibody response to rMxV in the EIA. A seroepidemiological survey of sera from 338 children in London showed that infection with MxV occurred earlier in life than NV. Primary infections with MxV were common after the age of 6 months. Over 70% of children had evidence of infection by the ages of 2 years, whereas only 12% of these children had been infected with NV. High concentrations of maternal antibody were present during the first month of life which was detected in 96% of the neonates. The results suggest that the high sensitivity of the EIA may be detecting maternal antibody throughout the first 8 months of life.

Adolescent↗
PubMed1995

Molecular characterisation of small round structured viruses associated with gastroenteritis in South Africa.

The application of the reverse transcriptase polymerase chain reaction (RT-PCR) has enabled several morphologically and physically similar small round structured viruses (SRSVs), including the prototype Norwalk virus (NV), to be classified within the Caliciviridae. This technique, using primers directed to the RNA-dependent RNA polymerase region within the ORF1 of NV, was used to characterise SRSVs associated with epidemic gastroenteritis in adults and sporadic paediatric gastroenteritis in South Africa. Genomic variation was investigated by sequence analysis of the amplified 209bp cDNA region from six isolates and comparison with other characterised SRSVs including NV. Antigenic variation was investigated by the use of the recombinant enzyme immunoassay described recently for the detection of Snow Mountain agent-like antigen in stool specimens. Two distinct antigenic groups were evident with NV-like viruses associated with adult gastroenteritis, and Mexico viruslike viruses associated with paediatric gastroenteritis. Viral isolates from two of the outbreaks of adult gastroenteritis showed a high degree of nucleotide sequence identity with NV, i.e., 84% and 98%, respectively, whereas the paediatric isolates showed 92-95% sequence similarity with the Snow Mountain-like virus, MxV. These data show concordance between antigenic and genomic analyses.

Adult↗
PubMed1995

Use of dried blood spots for the detection and confirmation of HTLV-I specific antibodies for epidemiological purposes.

AIMS--To modify and evaluate a gelatin particle agglutination test that could provide a sensitive, specific and inexpensive method for the detection of HTLV-I antibody in dried blood spot samples (DBS) collected on filter paper. METHODS--A set of 26 reference samples confirmed as HTLV-I antibody positive were assembled from patients with tropical spastic paraparesis or adult T cell leukaemia and blood donors. Serum samples and simulated antibody positive dried blood spot eluates were tested using the Serodia assay together with two confirmatory tests: HTLV BLOT 2.3, a western blot, and Select-HTLV, an enzyme immunoassay (EIA). Both confirmatory tests use synthetic peptides to differentiate between antibodies to HTLV-I and -II. The modified Serodia assay was then used to test anonymously 10,135 DBS collected from neonates from London. Samples reactive in the modified Serodia test producing a positive result were titrated to an end point and confirmed as before. RESULTS--All 26 eluates made from simulated DBS derived from positive reference samples were identified as positive by the modified Serodia HTLV-I test and were confirmed as anti-HTLV-I positive by EIA. Two eluates derived from relatively low titre reference samples gave indeterminate results on western blotting. Screening of the 10,135 neonatal DBS resulted in six repeat reactives, five of which were confirmed. The remaining reactive sample gave an indeterminate result on western blotting and there was insufficient eluate for testing by EIA. The overall seroprevalence of HTLV-I in this population was 0.05% (five of 10,135). CONCLUSION--The modified Serodia HTLV-I assay provides a sensitive, specific and inexpensive (10 pence/test) method for screening large numbers of DBS. The format of the assay makes it ideally suited for simultaneous screening of antibodies to HIV-1, HIV-2 and HTLV-I using semi-automated equipment.

Adult↗
PubMed1994

Seroprevalence studies using a recombinant Norwalk virus protein enzyme immunoassay.

A recombinant Norwalk virus (NV) protein enzyme immunoassay was used to study the age of acquisition of NV IgG in various populations. In London, England, there was little evidence of infection during the first 2 years of life. However, the prevalence of NV IgG rose steadily throughout the period that children attend school, reaching a peak of 70% in the group aged 11-16 years. High levels of maternal antibody were detected in infants aged < 3 months. Comparison of the acquisition of antibodies to three strains of human calicivirus in Japanese children in northern Japan indicated that although the majority had experienced infection with strains Japan and UK1 by the age of 12 years, only 22% possessed antibodies to NV. In Australian aborigines NV infection occurs early in life; by the age of 6 years over 90% of children were seropositive.

Adolescent↗
PubMed1994

Identification of adenoviruses in faeces from patients with diarrhoea at the Hospitals for Sick Children, London, 1989-1992.

Faecal samples from 137 patients that had been shown to contain adenoviruses by electron microscopy were identified in a series of enzyme immunoassays (EIA) using a single monoclonal antibody (Mab) to adenovirus 40 and four different Mabs to adenovirus 41. Adenoviruses were partially characterised by restriction enzyme analysis (REA) of DNA extracts using SmaI. Samples were also run in a commercial EIA (Adenovirus IDEIA; Dako, Ltd.) which detects group antigen. The majority (84%) of adenoviruses were subgenus F: adenovirus type 41, 87 (64%) and adenovirus type 40, 28 (20.4%). Subgenus A viruses were identified in ten, (7%) patients, eight were type 31, and two type 12. The adeno IDEIA test was sensitive and specific, detecting 127 of 131 positives and giving no false-positive results with other enteric viruses. Use of monoclonal-based EIAs showed significant differences depending on which adeno 41 Mab was used, although the restriction patterns obtained using SmaI appeared to be identical for 66 of 69 samples that produced recognisable bands. The Mab that performed best, M 4.3.1, was raised against strains obtained from children in England and detected 83 of 84 (99%) of the adenovirus 41 samples tested. In contrast Mab JH/41 raised against the prototype strain of adenovirus 41 (Tak) detected only 69 of 87 (79%).

Adenoviruses, Human↗
PubMed1994

Sequence similarity of human caliciviruses and small round structured viruses.

The application of reverse transcription-polymerase chain reaction (RT-PCR) using primers directed to the RNA dependent RNA polymerase region within ORF1 of Norwalk virus (NV) showed that 31 percent of morphologically typical human caliciviruses (HuCV) and 57% of small round structured viruses (SRSVs) produced a product of 470 bp similar to the NV control, NV 8FIIa/68/US. Alignment of the amino acid sequences of morphologically typical HuCVs with previously published sequences for SRSVs, NV, and Snow Mountain agent (SMA) showed a high degree of homology (90-92%) with SMA and a lesser extent of homology with NV (60-61%). The amino acid sequence of two strains of HuCV, HuCV/3C/92/UK, and HuCV/5C/92/UK differed by only one or two amino acids respectively in the RNA dependent RNA polymerase region from that of two strains of SRSV obtained from children in the United Kingdom, SRSV/4S/90/UK and Japan, SRSV/OTH-25/89/J which were found to have identical amino acid sequences. The use of an EIA for detection of NV antigen employing antisera raised to recombinant NV protein indicated that HuCVs and SRSVs obtained from children and adults in the United Kingdom were antigenically distinct from the prototype Norwalk virus, NV/8fIIa/68/US.

Adult↗
PubMed1994

Measurement of IgA responses following Norwalk virus infection and other human caliciviruses using a recombinant Norwalk virus protein EIA.

An enzyme immunoassay employing recombinant Norwalk virus capsid protein was evaluated for the measurement of IgA responses. Tests on 23 volunteers and patients known to have been infected with Norwalk virus (NV) showed that 19 developed significant IgA responses, 2 had unchanging levels of IgA and 2 failed to respond. There was no evidence of IgA responses to NV following infection with Hawaii or Snow Mountain-like viruses. Tests on sera from patients involved in outbreaks associated with eating contaminated shellfish suggest that some patients may have been infected with more than one strain of calicivirus. The use of the rNV EIA for measuring IgA and IgG responses in patients involved in a major outbreak of food poisoning affecting hospital staff indicated that the causative agent was probably NV.

Adult↗
PubMed1994

Cell culture adaptation of astrovirus involves a deletion.

Astroviruses have been adapted to culture by serial blind passage in primary human embryo cells. All viruses thus adapted possess a 45-nucleotide deletion relative to fecal viruses or isolates made in CaCo-2 cells; this deletion may be responsible for the change in host cell range.

Amino Acid Sequence↗
PubMed1994

Impact of rotavirus infection on a paediatric hospital in the east end of London.

AIMS: To study the impact of confirmed rotavirus infection at a paediatric hospital; to use the data to obtain a minimum estimate of the cost of treating reported cases of rotavirus in England and Wales. METHODS: Data were obtained on all patients with rotavirus over a two year period. Information was collected on 386 patients with rotavirus infection who were treated at the 120 bed Queen Elizabeth Hospital for Children in East London. This included the virus serotype, the patient's age, whether they required intravenous infusion, duration of hospital stay, numbers of patients treated in the casualty department, and numbers who had to be admitted. Treatment costs were obtained from the Finance Department of the Hospitals for Sick Children. RESULTS: The minimum cost of treating patients, excluding the cost of medical staff at the hospital, was estimated to be 95,400 pounds a year. One hundred and forty eight (38%) patients were admitted to the wards and a further 49 patients developed symptoms while in hospital. Intravenous infusion was required by 18 patients. The mean duration of hospital stay was 5.5 days. One hundred and eighty nine (49%) patients were treated with oral rehydration solution in casualty, given advice, and sent home. Ninety four per cent of the patients were aged under 2 years. The findings were comparable with those obtained in a study at Texas Children's Hospital, USA. The G serotype (VP7) of rotavirus did not influence the severity of infection. CONCLUSION: Rotavirus infections accounted for a significant number of patients treated in casualty, admissions to hospital, and bed occupancy in a paediatric hospital. The estimated cost of treating reported cases of rotavirus in England and Wales is in excess of 6.3 pounds million a year.

Age Factors↗
PubMed1993

Two successive outbreaks of SRSV-associated gastroenteritis in South Africa.

Two successive outbreaks of gastroenteritis in South Africa were investigated to identify the aetiological agents. Some patients were involved in both outbreaks. Enteropathogenic bacteria or parasites were not evident in either outbreak. Small round structured viruses (SRSVs) were demonstrated in both outbreaks by direct electron microscopy. SRSV UK3/Hawaii virus was identified by immune electron microscopy as the causative agent in the first outbreak. Using new recombinant Norwalk virus (rNV) immunoassays for antibodies and antigen, Norwalk virus was implicated in the second outbreak. Preexisting antibodies to Norwalk virus were not protective and there was no cross protection between Hawaii and Norwalk viruses. There was no anamnestic response to Norwalk virus following the SRSV UK3/Hawaii outbreak although those affected had preexisting antibodies to Norwalk virus. To our knowledge, this is the first definitive diagnosis of SRSV-associated gastroenteritis in South Africa.

Adult↗
PubMed1992

Modified latex agglutination test for antibodies to Toxoplasma gondii in eluates from Guthrie cards.

AIMS: To determine whether the Eiken particle agglutination test could be modified to make it sufficiently sensitive to screen blood samples collected on Guthrie cards for the presence of antibodies to Toxoplasma gondii; to evaluate the specificity of the modified system; and to compare seroepidemiological data on the prevalence of T gondii in pregnant women. METHODS: Simulated dried blood spots were prepared from sera from pregnant women booking for antenatal care. Eluates from the simulated dried blood spot cards and sera were tested in parallel using the modified test (1 in 5 dilution of latex) and the standard assay (neat latex particles) and endpoints determined. Guthrie card eluates, from neonates in three Thames regions, were then tested using the modified test. RESULTS: The modified test produced a 4.21-fold increase in antibody titre in 85 sera when tested in parallel with the standard test. Eluates of 168/170 from simulated dried blood spots derived from seropositive patients gave a positive result in the modified test. The two eluates which gave a negative result were derived from patients with an equivocal titre of 1/16 in the standard serum test. Of the eluates derived from serum negative patients all 103 were negative at a dilution of 1 in 4 in the modified test. The seroprevalence of antibodies to T gondii in pregnancy was 21.8% using the standard test. A similar value of 20.5% was obtained when dried blood spots from neonates in a similar region of London were tested by the modified test. CONCLUSIONS: The modified Eiken Toxo-reagent test is sensitive, simple, and economic for screening large numbers of dried blood spots. The procedure could be easily semiautomated and the technique applied to the mass screening of neonatal blood samples collected on Guthrie cards to determine the seroprevalence of T gondii in pregnant women.

Animals↗
PubMed1992

Evaluation of five commercial assays for screening antenatal sera for antibodies to Toxoplasma gondii.

AIMS: To evaluate the suitability of five commercial assays (Toxoreagent, DA, Captia Toxo IgG, Toxenz-G, Toxonostika-G) for screening large numbers of sera for antibodies to Toxoplasma gondii. METHODS: Sera from 1000 pregnant women booking for antenatal care at a London hospital were screened in parallel by each test. Sera giving discordant results were retested. RESULTS: The Captia Toxo IgG enzyme immune assay gave the best specificity on initial screening, with 0/773 false positives and only 2/218 false negatives. The Toxoreagent latex agglutination test performed well provided sera were tested at several dilutions to prevent prozone effects; 0/218 false negatives (greater than 12 IU/ml). Only one evidently false positive result was seen in the 1000 samples tested. The DA test gave no false negative results but produced 23/773 false positives. After repeat testing there were 9/1000 sera which gave equivocal results which were negative by the Captia Toxo IgG test (less than 12 IU/ml) but with low titres of 16 in the Toxoreagent test or 4 IU/ml in the DA test. In this situation women would have been asked for a follow up sample for repeat testing. Only 300 sera were tested by Toxenz-G; initial screening produced 4/58 false negative results and 4/242 false positives. CONCLUSIONS: The Captia Toxo IgG test gave the fewest discordant results on initial screening. Results could be readily expressed in international units using a programmable plate reader, and this may be useful for epidemiological studies. The Toxoreagent test is considerably cheaper, and is a simple and reliable method for screening provided that at least two dilutions are used.

Agglutination Tests↗
PubMed1991

Epidemiological survey of human rotavirus serotypes and electropherotypes in young children admitted to two children's hospitals in northeast London from 1984 to 1990.

A retrospective and prospective survey was carried out to determine the relative frequency of rotavirus serotypes infecting children with diarrhea or vomiting or both who were admitted to the Hospitals for Sick Children in London during a 6-year period from 1984 to 1990. The results were compared with data for the same period from a study in Birmingham, United Kingdom. The serotype of rotaviruses infecting 1,019 children was ascertained by enzyme immunoassay with VP7-specific monoclonal antibodies. In London, serotype G1 accounted for 60% of the cases, serotype G4 accounted for 24%, serotype G2 accounted for 11%, G3 accounted for 3%, and coinfections accounted for 2%. Considerable differences in the relative prevalence of serotypes were seen when data from London and Birmingham were compared. A major shift from serotype G1 to G4 was observed in London in the 1989 to 1990 season, and a lesser shift was seen in Birmingham. Examination of the electrophoretic profiles of 611 rotaviruses from London showed that there were at least 108 different profiles. Continuous variation occurred throughout the 6-year period, and the same electropherotype never recurred once it had disappeared from the population. None of the electrophoretic profiles were characteristic of group B or group C rotaviruses. There was no evidence that any strain of rotavirus had become endemic in either of the children's hospitals in London.

Child, Preschool↗
PubMed1990

Immunological characterization of the Marin County strain of astrovirus.

Marin County virus (MCV) was isolated from a stool suspension and serially propagated in human embryonic kidney cell cultures. MCV particles in stool and cell-propagated virus stocks showed reactivity by immune electron microscopy (IEM) with rabbit antiserum to astrovirus type 5. MCV antigen was also detected in two MCV stool samples by enzyme immunoassay (EIA) with an astrovirus group-specific monoclonal antibody. Acute and convalescent sera from 3 of 3 MCV-infected patients showed seroconversion to cell-propagated MCV by EIA. Immunofluorescence of MCV propagated in cell culture showed positive reactivity with an astrovirus group specific monoclonal antibody and astrovirus type 5 antiserum, with some cross-reactivity with astrovirus type 1. Similar results were obtained with the prototype strain of astrovirus type 5. However, in plaque-reduction assays, both the prototype astrovirus type 5 and MCV were neutralized by type 5 antiserum only. We conclude that MCV can be serially propagated by techniques used for previously described astroviruses and is serotypically an astrovirus type 5.

Antibodies, Monoclonal↗
PubMed1990

Human, small round structured viruses, caliciviruses and astroviruses.

Epidemiological studies on SRSVs, human calicivirus and astroviruses have been limited by the problems of establishing them in cell culture and the inability to transmit them to animals or to use strains from animals as a source of antigen for diagnostic tests. The use of EM and the subsequent development of RIAs and EIAs in a few research centres has shown that they are a cause of outbreaks and sporadic cases of diarrhoea and vomiting. SRSVs have increasingly been recognized as a major cause of outbreaks of gastroenteritis in the community and in hospital wards. The symptoms of illness are generally mild and of short duration and patients seldom require medical attention. However, because of the high attack rates and large numbers of persons of all age groups involved, there is often considerable economic loss and disruption of services. Evidence is accumulating that polluted water, molluscan shellfish, and contaminated cold foods are major sources of infection. Recently a SRSV has been shown to be the cause of epidemics and sporadic cases of waterborne enterically transmitted non-A, non-B hepatitis (hepatitis E virus) which have occurred in the USSR, India, Mexico and Africa. Astroviruses and human caliciviruses are occasional causes of outbreaks of vomiting and diarrhoea in infants and the elderly which can necessitate the closure of hospital wards and cause considerable disruption. Symptoms are generally mild and of short duration and therefore the majority of cases are unlikely to be investigated by laboratories. Diagnosis of infections is at present limited to the few laboratories that have developed their own assays or have access to electronmicroscopy facilities.

Adolescent↗