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Biomedical subjects

P P Mortimer

Publications and source records attributed to P P Mortimer.

At least 19 recordsLinked to original sources

Performance assessment of neonatal dried blood spot testing for HIV antibody.

A study was undertaken to assess the performance of laboratories participating in the UK unlinked anonymised neonatal screening programme for the detection of HIV antibody. A coded panel of 100 contrived dried blood spots prepared form well characterised anti-HIV-1 and anti-HIV-2 positive sera and an anti-HIV negative serum was distributed to eight testing centres. Fifty-one out of 52 anti-HIV-1 positive samples were identified by all eight laboratories: five laboratories identified all 52 specimens. Only one laboratory detected all four anti-HIV-2 positive spots. None of the laboratories reported a seronegative spot as reactive. The sensitivity of anonymised neonatal anti-HIV-1 screening was deemed satisfactory in all participating laboratories but the margin of sensitivity was insufficient to allow pooling of dried blood spot eluates before testing. Technical modifications would be necessary to increase sensitivity for anti-HIV-2 were HIV-2 infection to be reported more frequently in the UK. Performance assessment of the screening programme should be repeated at regular intervals.

AIDS Serodiagnosis

The detection in urine specimens of IgG and IgM antibodies to hepatitis A and hepatitis B core antigens.

The use of urine as a noninvasive specimen for the diagnosis of hepatitis A (HAV) and hepatitis B (HBV) virus infections was investigated. Specimens of urine were collected at the same time as blood or saliva specimens, or singly in cases of previously serologically confirmed recent infection. The specimens were tested for IgG and IgM anti-HAV and anti-HBc by immunoglobulin class-specific capture radioimmunoassays (GACRIA and MACRIA). On the basis of assays on urine specimens it was possible to distinguish between individuals who were susceptible or immune to HAV or who had recently been infected with HAV. Using assays on 327 corresponding saliva specimens as reference tests, the observed sensitivity and specificity of tests on urine specimens by anti-HAV GACRIA were 98.9% and 99.1%, respectively, and by anti-HAV MACRIA were 95.8% and 99.6%, respectively. IgM and IgG anti-HBc were detected readily in the urine of 35 acute or recent cases of hepatitis B but were not found in the urine of seronegative individuals. Of the urine specimens from 52 individuals who were HBsAg carriers or who had had long past HBV infections, 49 contained detectable IgG anti-HBc. Of urine specimens from 42 HBsAg carriers, 11 contained raised IgM anti-HBc levels. Urine, which is a convenient specimen to collect, can be used to study outbreaks of hepatitis A, to ascertain the HAV immune status of individuals, to differentiate hepatitis A from hepatitis B, and to identify individuals who have been naturally exposed to HBV.

Hepatitis A

Is the IEMA combined incubator/shaker device preferable to a waterbath for incubating enzyme immunoassays?

An incubator/shaker device proved to be a convenient alternative to a waterbath for the incubation of enzyme immunoassays (EIA). The device achieved effective and even heat transfer. In two of five EIAs it increased reactivity and in three of five EIAs it slightly increased the discrimination between seronegative and seropositive specimens though, for the samples investigated, extra sensitivity was not thereby achieved. At a high shaking frequency (1400 rpm) there was cross-contamination between wells, but this did not occur at 900 rpm. The relative contributions of heating and of shaking to the incubation of EIAs deserve further investigation.

AIDS Serodiagnosis

Confirmation of second generation anti-hepatitis C virus enzyme immunoassays by antigenic cross-validation.

AIM: To determine if a scheme for validating enzyme immunoassay (EIA) results could be devised that did not require costly and methodically elaborate supplemental assays. METHODS: Samples (n = 525) from patients with haemophilia A, leukaemia, and chronic liver disease and at increased risk of hepatitis C virus infection were tested by EIA-1 (Ortho Diagnostics), an assay which uses recombinant HCV fusion proteins as antigens, and by EIA-2 (United Biomedical), an assay based on synthetic HCV oligopeptide antigens. RESULTS: Samples (n = 193) were repeatedly reactive in both EIAs. Of these, 190 (98%) yielded reactivities in both of two supplemental assays used, one an immunoblot assay (RIBA) using recombinant HCV polypeptides similar to EIA-1 antigens, and the other a neutralisation EIA (EIA-2N) based on antigenic competition with HCV peptides similar to EIA-2 antigens. The three samples not reactive in supplemental tests exhibited low EIA optical density (OD) values (signal/cutoff ratios of less than 3). Hence, all specimens reactive and yielding high OD values in both EIAs were also reactive in supplemental assays. Twenty four samples were reactive in EIA-1 only and nine (38%) of these were reactive in RIBA. Fourteen of the 15 (93%) specimens reactive in EIA-1 but not RIBA were derived from patients with chronic liver dysfunction. Two samples were reactive in EIA-2 only, of which one was reactive in EIA-2N and none in RIBA. CONCLUSIONS: Compared with EIA-2, EIA-1 yielded more validated reactive samples and resulted in more non-validated reactivities. It is therefore suggested that for clinical diagnosis: (i) EIA-1 be used for anti-HCV testing and EIA-2 for validation of EIA-1 reactivities; (ii) samples concordantly reactive in EIA-1 and EIA-2 and displaying high OD readings be considered HCV antibody positive without supplemental testing; (iii) supplemental testing by RIBA be limited to samples reactive in EIA-1 but equivocal or unreactive in EIA-2 and those concordantly reactive but exhibiting low absorbance readings.

Hemophilia A

Human B19 parvovirus infection: an example of multiple pathogenic effects determined by differences in host susceptibility.

B19 infection offers some general lessons about human viruses and their possible effects on the human host, as follows: (1) Ubiquitous apparently benign viruses may have severe effects on a compromised host. The virus may be invariable but the host can have diverse susceptibilities. (2) B19 and some other human viruses (though for none is the evidence so clear as for B19) have narrowly targetted effects. The host cell of B19 is a specialised progenitor of mature red cells: impairment of the function of this cell by B19 may cause profound anaemia. (3) The 'normal' host response to B19 may also cause disease, though this is self limiting. (4) The effects of malfunction of the virus' target cell are exacerbated when the immune response is impaired by congenital or acquired immunodeficiency, immunosuppressive therapy or, in the case of the fetus, developmental immaturity that allows the virus to persist.

Adult

Detection of HIV-1 antibodies in blood specimens spotted on filter-paper.

Described are the results of an international collaborative study to evaluate the use of whole blood samples spotted on filter-paper (BSP) for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). BSP samples were collected from 40 patients at risk for HIV-1 infection and tested blindly using commercially available HIV antibody test kits, either specifically manufactured or modified for this purpose. Parallel serum samples were also collected, and the antibody reactivity was defined and confirmed by Western blot. The results demonstrate that recovery of antibodies from BSP samples after elution can be comparable to that from serum. Some kits can be easily adapted to test BSP samples, while others cannot. At present, detection of HIV antibodies in BSP samples should therefore be carried out using kits specifically manufactured for this purpose or by the development of a modified protocol using a panel of BSP and their corresponding serum specimens.

AIDS Serodiagnosis

HIV-2 in the United Kingdom--a review.

Human immunodeficiency virus type 2 (HIV-2) was first recognised in 1986 and subsequently the infection was shown to be widespread in West Africa. There have been few case reports from countries outside the African continent. In North America and Europe the highest number of infections have been in Portugal and France. Twelve HIV-2 infections have been identified in the United Kingdom (UK) and nine of the twelve had some connection with Africa, mostly West Africa; in three cases only "sub-Saharan Africa" was stated on the report, and one was from Mozambique. The other three HIV-2 infections were identified as follows: one in stored sera from a man who died in 1978, one in a child from Portugal who was diagnosed in the UK as having Acquired Immune Deficiency Syndrome (AIDS) in 1985, and one in a homosexual man who was tested unlinked and anonymously in London in 1987. It is not known how many of the total number of UK HIV-2 infections are represented by these twelve, but among large numbers of blood donors and people attending genitourinary medicine clinics the occurrence of infection was rare.

AIDS Serodiagnosis

The influence of dental status on the detection of IgG class anti-viral antibodies in human saliva.

An antibody-capture radioimmunoassay was used to measure levels of IgG class antibodies to rubella and hepatitis A viruses in serum and saliva of 30 edentulous, 30 partially dentate and 31 dentate individuals. The prevalence of seropositivity for rubella was 98.9 per cent and for hepatitis A 73.6 per cent. The serum reactivities were generally greater than those for saliva. There were 8 false-negative results for saliva out of the 182 tests performed, of which 4 were in the edentulous group, 3 in the partially dentate and 1 in the dentate group. For both rubella and hepatitis A virus antibodies the (geometric) mean ratios between the saliva and serum reactivities were similar across the three dental groups. The values for sensitivity, specificity and positive predictive value suggest that assay of saliva for antiviral IgG antibody is a satisfactory technique regardless of dental status.

Adult

Preliminary report: accurate assays for anti-HIV in urine.

Untreated urine specimens from 358 patients (344 attending genito-urinary medicine clinics, 14 haemophiliacs) and 353 blood donors were tested blind by a simple IgG-capture particle-adherence test (GACPAT) and a rapid IgG-capture enzyme-linked immunosorbent assay (GACELISA) for antibody to human immunodeficiency virus (anti-HIV). All 158 urine specimens from seropositive subjects were anti-HIV positive by GACPAT and 157 of them (99.4%) were positive by GACELISA. Tests on 553 urine specimens from seronegative subjects gave two repeatable false-positive reactions by GACPAT (0.4%) and none by GACELISA. By means of a modified procedure anti-gp160 was detected by commercial western blot in the urine of 44 of 45 seropositive subjects examined. IgG-capture assays will detect anti-HIV in unconcentrated urine and so allow a diagnosis in circumstances when blood sampling is impracticable.

Blotting, Western