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Biomedical subjects

A W Joss

Publications and source records attributed to A W Joss.

At least 37 records · Page 2Linked to original sources

Toxoplasma polymerase chain reaction on experimental blood samples.

A two-stage polymerase chain reaction (PCR) assay employing oligonucleotide primers from the B1 gene of Toxoplasma gondii was developed and assessed for sensitivity and specificity. It was able to detect T. gondii DNA from as little as one parasite/sample in mock-infected rat or mouse leucocyte preparations. Parasitaemia was also identified in animals at five stages between 16 and 66 h after infection with the virulent RH strain, and at 12 stages between 2 and 38 days after infection with the cyst-forming Beverley strain. In the latter case, PCR was more sensitive than animal culture. No cross-reactions were observed in samples containing various opportunist pathogens which may also be found in the blood of immunocompromised patients.

Animals↗

Use of the polymerase chain reaction to detect Toxoplasma gondii in human blood samples.

AIMS: To assess the value of detecting Toxoplasma gondii in human blood samples using the polymerase chain reaction (PCR). METHODS: Blood samples in lithium heparin were examined from 34 patients with suspected toxoplasmosis, and six healthy volunteers with or without the addition of doubling dilutions of toxoplasma tachyzoites. Products of a nested PCR, using oligonucleotide primers of the B1 gene, were analysed by electrophoresis and stained by ethidium bromide. The primary product was 194 base pairs in length; the nested products were 160 or 97 base pairs. RESULTS: When toxoplasma tachyzoites were added to the leucocytes of six different volunteers, eight to 16 parasites were detected by nested PCR in one experiment and one to four parasites in eight experiments. All nine experiments were negative in samples without added tachyzoites. Of 34 patients, PCR was negative in 13 with recent lymphadenopathy; nine with persisting IgM, including two pregnant patients; four with reactivated infection due to immunodeficiency; and five with no evidence of active infection. Positive PCR results were found in three patients with reactivated infection. There was only one discrepancy between PCR and animal culture results; this was in an immunocompromised patient with a positive PCR and negative culture. CONCLUSIONS: The PCR technique was rapid, sensitive, and specific in human blood samples. Negative PCR results in patients with persisting IgM suggested that the fetus was not at risk, or that treatment was not indicated in myalgic encephalomyelitis-like illness. PCR results in immunocompromised patients permitted appropriate management--no treatment if negative, treatment if positive.

Animals↗

Comparison of relative uses of commercial assays for Toxoplasma gondii IgM antibodies.

AIMS: To compare the sensitivity and user friendliness of seven commercially available enzyme linked immunoabsorbent assay (ELISA) kits for toxoplasma specific IgM. METHODS: Five antibody capture assays supplied by Abbott, Mercia, Northumbria, Organon and Sorin, and two indirect ELISA assays from Biostat and Mast, were assessed. Using defined dilutions of Toxoplasma gondii specific IgM, the performance and sensitivity of each assay was established. They were further assessed on a panel of 27 sera with a range of dye test and IgM results (as determined by the Scottish Toxoplasma Reference Laboratory). All of the assays were performed by three experienced operators and assessed for user satisfaction. RESULTS: The Mast, Organon, and Abbott assays were of low sensitivity; the Mercia and Northumbria were of high sensitivity; and the Biostat and Sorin assays produced too many false positive results. The Mercia kit provided most user satisfaction; the Mast and Abbott assays were most difficult to use. CONCLUSIONS: Local laboratories investigating toxoplasma infection should have three tests: one IgG and two IgM (high and low sensitivity) to help in the timing of infection. Alternatively, one sensitive IgM assay, such as that of Mercia, could be used by selecting appropriate high and low thresholds.

Animals↗

Congenital toxoplasmosis: to screen or not to screen?

We have reviewed the present day quantifiable cost to society of the 73 cases of congenital toxoplasmosis which are estimated to occur annually in Scotland with the cost of preventing the disease by screening and treatment. Our analysis includes advances in laboratory techniques. The cost of screening would depend on its scale and if in-house or commercial tests are used. If only 2 specimens were screened, at booking and at delivery, the screening costs are estimated to be between 0.5-0.9 times the preventable costs. If a third specimen were tested in the second trimester, to maximise scope for remedial action during pregnancy, the screening costs are 0.7-1.2 times preventable costs. As likely screening costs in most of the schemes we consider are now less than the preventable costs, a screening programme should be adopted.

Cost-Benefit Analysis↗

Diagnosis of congenital toxoplasmosis in Scotland.

During a period of 10 years (1976-1986) 853 cases of toxoplasmosis were identified serologically in the Toxoplasma Reference Laboratory in Scotland. Suspected cases of infection in pregnancy (29 cases) and of congenital infection (27 cases) were rare but ophthalmic disease was common (224 cases). In two of the congenital cases, maternal infection was identified during pregnancy. The other 25 babies were investigated because of suspected congenital infection. Toxoplasmosis was confirmed retrospectively in 11/25 babies and excluded in two of them. The other 12 cases remain unclassified. Diagnosis of infection in babies was difficult although the introduction of an IgM capture ELISA was of some help. IgM was detected by ELISA in 3/13 congenitally infected babies but by immunofluorescence tests in only one of these. In two antenatal studies, involving 16,000 women, an additional 32 women were identified whose babies might have benefited from treatment or the availability of fetoscopy for early detection of congenital infection.

Animals↗

The use of an IgM immunosorbent agglutination assay to diagnose congenital toxoplasmosis.

An IgM immunosorbent agglutination assay (ISAGA) was compared with a standard ELISA IgM test for the diagnosis of congenital toxoplasmosis. It was more sensitive, detecting all of five mothers of infected babies whereas the IgM ELISA was positive in two of three mothers tested at delivery and neither of two mothers referred 10-12 months after delivery. Five women infected in a previous pregnancy had IgM detectable by ISAGA in a subsequent pregnancy. The assays were comparable when sera from patients with past infection were tested or following toxoplasma-associated miscarriage or abortion. Four cord sera from congenitally-infected babies were positive by the ISAGA but only three of these were positive by ELISA for IgM. The ISAGA also detected IgM in another four sera from congenitally-infected babies referred late (10-18 months old); none were IgM positive by ELISA. The increased sensitivity of the ISAGA is an improvement in the diagnosis of congenital toxoplasmosis.

Agglutination Tests↗

Biotin-labelled antigen screening test for toxoplasma IgM antibody.

A method for the simple preparation of biotin-labelled toxoplasma antigen was used with avidin peroxidase in an IgM-capture enzyme linked immunosorbent assay (BAM-ELISA). Although the overall predictive value of a positive result was only 38%, its low cost and 100% sensitivity makes it a very suitable screening test. Positive results can be confirmed by an alternative assay, thus providing a more economical and effective diagnostic service than either screening all sera by a commercial test or selecting sera for IgM testing.

Animals↗

Pneumocystis carinii antibody testing.

Sera from blood donors and patients from all over Scotland were examined by indirect immunofluorescence using Pneumocystis carinii antigen from infected rat lung. Antibody was found in 76 of 488 (15.6%) of patients tested on clinical grounds but in only 13 of 148 (8.8%) blood donors. The antibody rates were higher in disease groups likely to have or develop P carinii pneumonia: in those with histologically confirmed or strongly suspected P carinii pneumonia the rate was 14 of 24 (58.3%); in those who had undergone transplantation eight of 24 (33.3%); in those who were immunosuppressed five of 16 (31.2%); in those who were human immunodeficiency virus antibody (HIV) positive 11 of 43 (25.6%); in those with malignancy 34 of 233 (14.6%); and in those with chest infection 10 of 85 (11.7%). P carinii pneumonia was confirmed or likely in four of 45 (8.8%) patients with titres of 1/8-1/16 and in three of seven (42.8%) in those with titres of greater than or equal to 1/128. Seroconversion or rising titre was detected in seven of 13 (53.8%) cases of confirmed or likely P carinii pneumonia compared with 10 of 93 (10.7%) in other patients. Diagnosis of P carinii infection can therefore be assisted by positive immunofluorescence results, but negative serology does not exclude infection.

Animals↗

Toxoplasmosis: effectiveness of enzyme immunoassay screening.

The effectiveness of IgG enzyme-linked immunosorbent assays as serological screening tests for Toxoplasma gondii infection was investigated in three Scottish hospital laboratories. Three assays--two commercial and one produced by the Scottish Toxoplasma Reference Laboratory--were investigated. There was little difference in the sensitivity of the three tests (86-91%), but one of the commercial tests was markedly less specific. The performance of the more specific assays was improved by adjustment to an eight international unit per ml threshold.

Animals↗

Further investigations of a hydatid focus in North-West Scotland.

A serological survey carried out on 1379 school-children from Skye and Mallaig in the N.W. of Scotland failed to show evidence of subclinical cases of hydatidosis. There appeared to be a relatively low level of exposure to the parasite, as judged by ELISA and IHA tests. Thirty-one children reacted positively in the ELISA; only one child had a transient positive result in the IHA test. Surveillance of these children will be continued for the present but the duration required is uncertain. The local population appeared to be well aware of the mode of spread of the parasite and the methods for its control.

Animals↗