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Biomedical subjects

D P Casemore

Publications and source records attributed to D P Casemore.

At least 19 recordsLinked to original sources

Managing a large outbreak of cryptosporidiosis: how to investigate and when to decide to lift a 'boil water' notice.

The largest outbreak of cryptosporidiosis reported in the United Kingdom, involving 575 confirmed cases (of which 474 met an agreed case definition), occurred in the county of Devon during August and September of 1995. The descriptive epidemiology supports the hypothesis that the outbreak was associated with the consumption of cold tap water in the area served by a particular water treatment works. Cryptosporidium oocysts were detected in treated water samples at the time of the outbreak. Although the epidemiological analysis provided strong circumstantial evidence of a waterborne outbreak, the data were not recorded in a manner that made them admissible in criminal proceedings taken by the Drinking Water Inspectorate against the water company involved. The need to carry out an analytical study in conjunction with the identification and characterisation of the pathogen in the drinking water and the practicalities of agreeing criteria for lifting a 'boil water' notice are discussed.

Adolescent↗

PCR-ELISA: a new simplified tool for tracing the source of cryptosporidiosis in HIV-positive patients.

Cryptosporidium parvum is a major parasitic cause of death in end-stage AIDS patients that results from both zoonotic and person-to-person transmission. Recent studies have provided evidence that parasites causing zoonotic disease and those causing anthroponotic infection are genetically distinct. Isolates carrying "animal"-type genetic markers were presumed to be the result of zoonotic spread, either directly or through contaminated food and water. The need for a genotype-specific diagnostic tool that can provide clues as to the origin and possible modes of spread of C. parvum strains has been recognised. Here, we report the development of such a tool for C. parvum based on polymerase chain reaction-enzyme linked immunosorbent assay that enables the accurate typing of isolates from HIV-seropositive and HIV-negative patients presenting with diarrhoea from the United Kingdom and Canada. This study also showed that zoonotic transmission might be predominant in the HIV-positive patient group in the United Kingdom.

AIDS-Related Opportunistic Infections↗

Modification of a rapid method for the identification of gene-specific polymorphisms in Cryptosporidium parvum and its application to clinical and epidemiological investigations.

The application of genotyping to clinical isolates of Cryptosporidium has increased significantly our knowledge and understanding of the distribution and epidemiology of this parasite. However, some methods can be laborious and demand specialist technical expertise. PCR-restriction fragment length polymorphism (RFLP) techniques represent a more rapid and simple method of genotyping to support epidemiological and clinical investigations than conventional DNA analytical techniques. We describe a nested PCR-RFLP technique that identifies polymorphisms in the C. parvum thrombospondin-related adhesive protein gene locus; this method offers a sensitive and specific tool for the confirmation and investigation of disease associated with C. parvum. The potential of this enhanced method is demonstrated by its application to the confirmation and epidemiological investigation of an outbreak of cryptosporidiosis associated with a school visit to an open farm.

Adult↗

Outbreaks of waterborne infectious intestinal disease in England and Wales, 1992-5.

Following the introduction of an improved surveillance system for infectious intestinal disease outbreaks in England and Wales, the Public Health Laboratory Service Communicable Disease Surveillance Centre received reports of 26 outbreaks between 1 January 1992 and 31 December 1995 in which there was evidence for waterborne transmission of infection. In these 26 outbreaks, 1756 laboratory confirmed cases were identified of whom 69 (4%) were admitted to hospital. In 19 outbreaks, illness was associated with the consumption of drinking water from public supplies (10 outbreaks) or private supplies (9 outbreaks). The largest outbreak consisted of 575 cases. In 4 of the remaining 7 outbreaks, illness was associated with exposure to swimming pool water. Cryptosporidium was identified as the probable causative organism in all 14 outbreaks associated with public water supplies and swimming pools. Campylobacter was responsible for most outbreaks associated with private water supplies. This review confirms a continuing risk of cryptosporidiosis from chlorinated water supplies in England and Wales, and reinforces governmental advice to water utilities that water treatment processes should be rigorously applied to ensure effective particle removal. High standards of surveillance are important for prompt recognition of outbreaks and institution of control measures. As microbiological evidence of water contamination may be absent or insufficient to implicate a particular water supply, a high standard of epidemiological investigation is recommended in all outbreaks of suspected waterborne disease.

Communicable Diseases↗

The epidemiology of cryptosporidiosis: application of experimental sub-typing and antibody detection systems to the investigation of water-borne outbreaks.

A technique based on the analysis of banding patterns obtained by SDS-PAGE Western-blotting of an oocyst wall antigen obtained from faeces has been evaluated to subtype Cryptosporidium parvum Tyzzer, 1912. This technique appears to have sufficient stability to recognise multiple types of this parasite. A similar Western-blotting technique has also been used to assess antibody responses to cryptosporidial antigens in human sera. Two systems were developed: one against three antigens of apparent molecular weights 6, 14 and 17 kDa; the second against oocyst wall antigens of apparent molecular weights 57, 69, 75, 89, 128, 151 and 173 kDa. Antibodies to three antigens of apparent molecular weights 6, 14 and 17 kDa were most successful as diagnostic markers in that they were found in > 88% of convalescent phase sera from confirmed cryptosporidiosis patients and were uncommon (> or = 7%) in control subjects. Faecal samples from human and animal sporadic cases yielded a wide range of cryptosporidial antigen banding patterns. Samples from patients in a water-borne outbreak in South Devon (England) in 1995 also yielded a wide range of banding patterns including members within individual household family groups. These results are in contrast with those from samples collected from other defined geographical areas, including some from a second water-borne outbreak where much more homogeneous banding patterns were obtained. Sera collected for other purposes from apparently uninfected individuals 9 months after the South Devon 1995 outbreak were examined. Antibodies to the three antigens of molecular weights 6, 14 and 17 kDa were detected in 32-49% of individuals resident in the outbreak water supply area, and in 15-21% of those resident in an adjacent water supply area. The significance of these findings is discussed in relation to data obtained from epidemiological field studies.

Animals↗

Molecular characterisation of Cryptosporidium parvum from two large suspected waterborne outbreaks. Outbreak Control Team South and West Devon 1995, Incident Management Team and Further Epidemiological and Microbiological Studies Subgroup North Thames 1997.

Polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis of the Cryptosporidium parvum outer wall protein (COWP) gene was applied to specimens collected from 95 patients with cryptosporidiosis associated with two suspected waterborne outbreaks, 46 sporadic human cases and 62 infected livestock from other areas, and 12 patients infected with other gastrointestinal parasites. Ninety-six per cent of C. parvum isolates from patients linked to the two suspected waterborne outbreaks were of genotype 1; all the isolates from livestock were of genotype 2. Isolates from 59% of the sporadic human infections were of genotype 1 and 35% were of genotype 2. Specimens from two patients yielded both genotypes. Specimens from patients infected with other parasites yielded no amplicons.

Animals↗

A simple SDS-PAGE immunoblotting technique using an enhanced chemiluminescence detection system to identify polyclonal antibody responses to complex cryptosporidial antigen preparations following a monoclonal antibody retest and image overlay technique.

SDS-PAGE immunoblotting is a standard immunochemical technique. We describe a modification of this technique to facilitate the identification of polyclonal antibody responses to complex antigen preparations. The technique utilises an enhanced chemiluminescence (ECL) detection technique where an immunoblot is initially tested with polyclonal antibodies. The polyclonal antibodies are 'stripped' from the nitrocellulose support and the immunoblot is retested with a monoclonal antibody. Polyclonal antibody responses to individual components also recognised by the monoclonal antibody are identified by superimposing the two photographic-film images. This technique has the advantage that antigens can be identified across an individual blot despite distortions which may have occurred. An example of the use of this technique is described in which antibody responses in human sera against antigens from the coccidian parasite Cryptosporidium are identified.

Animals↗

PCR-RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between C. wrairi and C. parvum, and between C. parvum isolates of human and animal origin.

Cryptosporidium wrairi was isolated from guinea pigs during a spontaneous outbreak of cryptosporidiosis. Despite the morphological and antigenic similarities to C. parvum, C. wrairi displayed a different host range and site of infection and may represent a separate species or sub-species. We used the polymerase chain reaction to clone two distinct 550 bp-long DNA fragments, Wc-I and Wc-II, of the gene encoding the Cryptosporidium oocyst wall protein (COWP) of C. wrairi, which showed 98% identity to the C. parvum homologue. Within Wc-I, polymorphic Rsal restriction sites were used to develop a polymerase chain reaction-restriction fragment length polymorphism method able to distinguish C. wrairi from C. parvum and to identify two groups of C. parvum isolates differentially associated with animal and human infections.

Amino Acid Sequence↗

Detection of a picobirnavirus associated with Cryptosporidium positive stools from humans.

A picobirnavirus with an atypical genome profile was detected by polyacrylamide gel electrophoresis (PAGE) in 37% (20/54) of human faecal samples also containing oocysts of Cryptosporidium typical of C. parvum. This virus shares many of the characteristics of the previously described picobirnaviruses, but has a significantly smaller genome (1.75 and 1.55 Kbp).

Animals↗

An outbreak of waterborne cryptosporidiosis associated with a public water supply in the UK.

In November and December 1992, an outbreak of waterborne cryptosporidiosis occurred in Bradford, a city in the North of England. In all 125 cases were involved, the majority of whom lived in part of the city which received its drinking water supply from a single water treatment works. A case control study demonstrated an association between illness and the consumption of tapwater from this source; those drinking large volumes being more likely to have been ill. Treated water from the defined source yielded cryptosporidial oocysts. Heavy rainfall was recorded in the catchment area of the reservoir supplying raw water to the treatment works, immediately prior to the probable time of infection, based on dates of onset of illness.

Adolescent↗

Cryptosporidiosis associated with a swimming pool complex.

Twelve children and one adult in Gloucestershire fell ill with cryptosporidiosis in March 1992. Ten cases lived in or near a particular Cotswold town. Eight of these had visited a swimming pool within 24 hours of a suspected faecal accident, and two may have been secondary cases. Of the three cases who lived elsewhere, one was probably unrelated to the outbreak, one had visited the pool, and one was a contact of a boy who may have been the source of the outbreak.

Adult↗