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J R Pattison

Publications and source records attributed to J R Pattison.

At least 19 recordsLinked to original sources
PubMed1992

Human parvovirus B19 in clotting factor concentrates: B19 DNA detection by the nested polymerase chain reaction.

The presence of B19 parvovirus in plasma from blood donors is seldom demonstrable, but clotting factor concentrates, prepared from large plasma pools, may be able to transmit B19 virus infection, and the effectiveness of different chemical and physical treatment to inactivate this virus is not yet known. In this study we report on the detection of B19 DNA in 25 clotting factor concentrates, prepared by a variety of procedures of purification and inactivation; dot blot hybridization and Southern blot hybridization assays, as well as a 'nested' polymerase chain reaction (PCR) have been employed. Nine out of 25 products were B19 DNA positive by PCR, whereas only two gave positive results by hybridization techniques. B19 DNA positive concentrates have been found in 'untreated' products but also in some solvent/detergent or steam-treated products and even in monoclonal purified concentrates. PCR may be useful for the screening of blood products to be used in immunocompromised haemophiliacs, particularly in HIV positive subjects, at risk of severe chronic anaemia following B19 infection.

Base Sequence↗
PubMed1991

Prevalence of antibodies against parvovirus B19 in Norwegians with congenital coagulation factor defects treated with plasma products from small donor pools.

The seroprevalence of antibodies against parvovirus B19 in 308 Norwegians with coagulation factor defects of different types and severities was assessed by an IgG antibody capture radioimmunoassay (GACRIA). The overall seroprevalence was 62%. The seroprevalence among subjects with different types of coagulation factor defects was related to the type and severity of the coagulation factor defect: severe hemophilia A 64%, moderate and mild hemophilia A 58%, severe hemophilia B 88%, moderate and mild hemophilia B 73%, and von Willebrand's disease 52%. The prevalence of parvovirus B19 antibodies among household contacts and blood donors was 49% and 42% respectively. This study confirms that replacement therapy with coagulation factors is accompanied by an increased risk for acquiring parvovirus B19 infection. However, the prevalence of parvovirus B19 antibodies among Norwegian hemophiliacs is well below the prevalence reported from other countries and probably reflects the small numbers of donors in plasma pools used for the preparation of coagulation factor concentrates.

Adult↗
PubMed1990

The production of human parvovirus capsid proteins in Escherichia coli and their potential as diagnostic antigens.

We have expressed a number of polypeptides derived from the capsid proteins of the human parvovirus B19 in Escherichia coli. These include native VP1 (84K) and VP2 (58K) proteins and also fusions to beta-galactosidase containing differing amounts of the amino terminus of the VP1/2 polypeptide. Although each of these was expressed at high levels and the majority were produced as full-length proteins, only one was soluble. This soluble polypeptide, p132, is a beta-galactosidase fusion protein that includes 145 amino acids from B19 which are entirely derived from the region unique to VP1. Despite containing such a small portion of VP1, which itself constitutes only 4% of total capsid protein, p132 reacted with all our known anti-B19 IgM-positive human serum samples. We conclude that this region contains epitopes which must be prominently exposed on the intact virus. We have demonstrated the use of this recombinant antigen in a simple diagnostic assay for B19-specific antibodies which can be used for initial screening of human serum samples. In a survey of 103 serum specimens, our ELISA positively identified all samples (19/19) which were positive by IgM antibody capture radioimmunoassay. The recombinant p132 antigen is efficiently produced and readily purified from E. coli, and its use as a diagnostic antigen should increase the availability of routine clinical testing for human parvovirus infection.

Antigens, Viral↗
PubMed1990

The pathogenesis of diseases associated with B19 virus.

B19 virus infection is common in the population and is frequently asymptomatic. However, a viraemia and prompt antibody response occurs in normal individuals and this is associated with mild, non-specific respiratory tract symptoms at the time of the viraemia and/or a rash-illness a week or ten days later. Infection of red cell precursors is a regular occurrence and this leads to aplastic crisis if B19 virus infection occurs in an individual with chronic heamolytic anaemia. Fetal infection sometimes takes place if infection occurs during pregnancy and some fetuses fail to clear the infection, develop anaemia leading to heart failure and hydrops fetalis. Some immunocompromised patients also fail to clear the viraemia and this results in a persistent or relapsing anaemia.

Female↗
PubMed1979

Comparison of immunofluorescence and radioimmunoassay for detecting IgM antibody in infants with the congenital rubella syndrome.

Immunofluorescence (IF) and radioimmunoassay (RIA) have been compared as methods for detecting IgM antibody in 124 infants with confirmed or suspected congenital rubella. IF was used to test sucrose density gradient fractions and RIA to test fractions and whole serum. When fractions were tested IF and RIA were equally specific and distinguished clearly between IgM and IgG, but RIA was the more sensitive method. The RIA titre in whole serum was always greater than in the peak IgM fraction and there was no evidence that testing the serum, rather than the fraction, could result in failure to detect IgM. With some sera RIA gave low titres which became negative after absorption with IgG-coated latex beads. The mechanism of this 'false positive' effect, which may have been due to IgM with anti-IgG activity, was not investigated, but if it can be removed by absorption it need not reduce the specificity of the test. During the first 6 months of life IgM antibody was detected by RIA in 30 out of 32 unfractionated sera and by IF in fractions from 28 of these. After the age of 6 months IgM was found progressively less frequently and the greater sensitivity of RIA became a more obvious advantage: 17 out of 60 specimens were positive by RIA and 11 of these were negative by IF. RIA testing of whole serum appears to be an economical, specific and sensitive method for detecting IgM antibody in congenital rubella, of particular value when the titre of antibody is low.

Age Factors↗
PubMed1979

A comparison of three tests for rubella antibody screening.

Sera from 1000 women attending antenatal clinics were screened for the presence of rubella antibody by haemagglutination inhibition (HI), single radial haemolysis (SRH), and enzyme-linked immunosorbent assay (ELISA). With 925 sera concordant results were obtained with all three tests. There were 46 false-negative HI tests due to the necessity of allowing for residual non-specific inhibitors. With SRH there was one false positive and three that were negative by SRH but HI- and ELISA-positive. By ELISA 14 sera gave a false-positive result (OD 405 nm greater than or equal to 0.2). Ten of these could be eliminated by taking an OD 405 nm of 0.5 as the threshold but then another 10 sera became false negatives.

Antibodies, Viral↗
PubMed1979

The nature of the effector cells of cell-mediated immune responses to Sendai and Kunz virus infections in mice.

The development of virus-specific cell-mediated immune (CMI) responses to Sendai and Kunz virus infections of the respiratory tract of mice is described. In addition a heterotypic cell-mediated immune response develops during each of the virus infections. Cell depletion studies indicate that the effector cells of the virus-specific CMI responses are theta antigen bearing T lymphocytes. Similar studies of the heterotypic responses suggest that 2 cell types are involved, T lymphocytes and NK cells.

Animals↗
PubMed1978

The use of high speed gel filtration for the detection of rubella-specific IgM.

Three new gels, Ultrogels AcA 22, AcA 34 and AcA 44 have been compared with Sephadex G-200 for the detection of rubella-specific IgM. Ultrogel AcA 44 did not achieve an adequate separation of IgM and IgG. Ultrogels AcA 22 and AcA 34 gave clear separation of IgM and IgG but only the latter could be operated at flow rates significantly higher than Sephadex G-200. With rubella convalescent sera fractionation with Ultrogel AcA 34 leads to a 2-4-fold loss of sensitivity for the detection of the first peak of rubella HI activity which is associated with recent infection. This is largely due to the occurrence of specific IgA antibody in the first peak of rubella HI activity after Sephadex G-200 fractionation but not after Ultrogel AcA 34 fractionation.

Antibody Specificity↗
PubMed1978

The derivation of a minimum immune titre of rubella haemagglutination-inhibition (HI) antibody. A Public Health Laboratory Service collaborative survey.

Ten laboratories collaborated in a study of minimum immune titre (MIT) of rubella haemagglutination-inhibiting (HI) antibody with one laboratory acting as a reference laboratory to provide a uniform basis for comparison of the HI results. The international unitage equivalent to the MIT used by the ten laboratories was found to vary from 24 to 98 units. Testing of the sera by immunofluorescence and by HI after flotation centrifugation indicated that residual non-specific inhibitors may interfere with HI antibody testing to an extent equivalent to 12-15 units. An acceptable MIT would therefore be equivalent to 24-48 units of rubella HI antibody. The single radial haemolysis (SRH) results on the sera indicate that this is a sensitive and specific test for rubella antibody.

Antibodies, Viral↗
PubMed1978

Comparison of methods for detecting specific IgM antibody in infants with congenital rubella.

Serum specimens from 14 infants with congenital rubella were examined for specific IgM antibody by six different methods. IgM-containing fractions were separated either by sucrose density-gradient centrifugation or by gel filtration through Sephadex G-200, and were then tested by the indirect immunofluorescence technique and by the haemagglutination-inhibition (HI) test (long-and short-incubation methods). Immunofluorescence staining of density-gradient fractions detected specific IgM in all 14 infants. The HI test (long method), applied to density-gradient fractions, was almost as sensitive, detecting antibody in 13 infants; the short method was less sensitive. The gel-filtration technique proved to be generally less satisfactory than sucrose density-gradient centrifugation. Evidence was obtained for the occurrence of as yet unclassified non-specific inhibitors in the serum of some infants. These inhibitors were deposited with the IgM on sucrose-density gradients and they could have been mistaken for rubella-specific IgM antibody, particularly in the HI test (long method).

Antibodies, Viral↗
PubMed1978

The detection of rubella-specific IgM antibodies by radioimmunoassay.

An indirect solid-phase radioimmunoassay (RIA) has been developed for the detection of immunoglobulin (Ig) class-specific rubella antibodies. A commercial rubella haemagglutinin is dried and fixed on to the wells of flexible microtitre plates and allowed to react with serial dilutions of whole or fractionated human sera. Class-specific rubella antibodies are then detected by determining the specific binding of 125I-labelled anti-human IgG or IgM. The RIA was first evaluated by comparison with the haemagglutination-inhibition (HI) test for the detection of rubella-specific IgM in gel-filtration fractions. RIA was found to be as specific as HI but 10-150 times more sensitive. Rubella-specific IgG antibodies did not interfere in specific IgM determinations by RIA and therefore the latter technique was applied to unfractionated sera. The results obtained indicate that RIA on unfractionated sera is a practical, sensitive and specific technique which could provide a reliable method for the diagnosis of rubella. The rubella-specific IgM titres obtained by RIA were not increased by the removal of IgG by pretreatment of sera with Staphylococcal Protein A.

Antibodies, Viral↗
PubMed1977

Cell-mediated immunity to Sendai virus infection in mice.

The development of a cell-mediated immune response to Sendai virus infection in mice was examined by the use of a 51Cr release assay of cytotoxicity. A low level of "background cytotoxicity" to Sendai virus-infected L cells was found in the spleens of uninfected CBA mice. Spleen cells from Sendai-infected mice showed an elevated level of cytotoxicity against these target cells for a period of 5 weeks, commencing 4 days after infection of the mice. A more transient response was observed in the spleens of mice infected with a serologically distinct virus, the Kunz strain of influenza. This cross-reacting, cell-mediated immune response was intermediate between that observed in unsensitized and Sendai-sensitized spleen cells. The relevance of these cell-mediated immune responses to respiratory tract virus infections is discussed.

Animals↗
PubMed1977

Serum IgM antibody and influenza A infection.

Sucrose density gradient ultracentrifugation followed by haemagglutination inhibition for demonstrating specific influenza IgM was evaluated as a means of confirming recent infection with influenza A viruses. Specific IgM antibodies were found in at least one serum obtained from 83% of patients with proven recent infection with influenza A viruses but in none of the sera from 21 individuals without evidence of infection. Influenza IgM antibodies persisted for up to 112 days after infection. The relative merits of detecting specific IgM and complement fixing antibodies for diagnostic purposes are discussed.

Antibodies, Viral↗
PubMed1976

Virus disease.

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Humans↗