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Automated reading and processing of quantitative IgG, IgM, IgA, and IgE isotypic agglutination results in microplates. Development and application in parasitology-mycology.

Microplate agglutination techniques represent a simple and commonly used approach for the quantitative or qualitative isotypic analysis of specific antibodies. However, they require optical reading by the investigator and are thus prone to an important degree of variability. In order to solve some of the problems associated with the variability of optical readings, we have used an automatic reader scanning each of the 96 wells of a standard microplate in 32 different locations. The inherent advantages of the automatic reader were further maximized by coupling it to a dedicated computer running customized software designed to process data coming on-line from the spectrophotometer. This approach has been applied to the diagnosis of human toxoplasmosis and candidosis. Suspensions of Toxoplasma gondii tachyzoites or of sensitised erythrocytes were used for the determination of IgG antibodies or the quantification of IgM, IgA, or IgE specific isotypes. This procedure allows the simple and reproducible collection of objective results. Moreover, it permits a reduction in cut-off values and direct interpretation of results with automatic conversion of scores into titer, units, index, or into any other scale appropriate for standardization purposes.

Agglutination Tests↗

A comparison of parasitological methods for the diagnosis of gambian trypanosomiasis in an area of low endemicity in Côte d'Ivoire.

The card agglutination test for trypanosomiasis (CATT) was used to examine 8974 inhabitants in 14 village areas south-west of Daloa, Côte d'Ivoire; 114 (1.3%) were CATTT or +/-, and were further examined by one or more of 6 methods for the direct detection of trypanosomes: lymphatic gland puncture, stained thick blood film (TBF), haematocrit centrifugation technique (HCT), mini-anion exchange column (MAEC), quantitative buffy coat method (QBC), and kit for in vitro isolation of trypanosomes (KIVI). Trypanosomes were seen by at least one method in 16 (14.0%) of the CATT+ group. Blood from 356 of the 8860 CATT- group was inoculated into KIVI; trypanosomes grew from the blood of 1 person. Eleven of the 17 patients with detectable trypanosomes were screened by all 6 methods: 6 were HCT+; 7 were gland+; 10 were MAEC+; 10 were KIVI+; 11 were both TBF+ and QBC+. One CATT+ patient was KIVI+ but otherwise negative, although TBF was not done. The overall prevalence of trypanosomes was 0.2% rising to 0.8% in one village area. The results support previous evidence that a reappraisal of procedures is required in the customary system of surveillance for gambian sleeping sickness.

Agglutination Tests↗

World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines for evaluating the efficacy of equine anthelmintics.

These guidelines have been designed to assist in the planning, operation and interpretation of studies which would serve to assess the efficacy of drugs against internal parasites of horses. Although the term anthelmintic is used in the title and text, these guidelines include studies on drug efficacy against larvae of horse bot flies, Gasterophilus spp, which are non-helminth parasites commonly occurring in the stomach of horses. The advantages, disadvantages and application of critical and controlled tests are presented. Information is also provided on selection of animals, housing, feed, dose titration, confirmatory and clinical trials, record keeping and necropsy procedures. These guidelines should assist both investigators and registration authorities in the evaluation of compounds using comparable and standard procedures with the minimum number of animals.

Animals↗

Uses and abuses of statistical methods in research in parasitology.

Among misuses of statistical procedures, perhaps most common is abuse of methods for multiple comparisons of means. Ranking close behind is misunderstanding of various aspects of analysis of non-random repeated measurements of animals. Those topics are the chief focus of this effort to do more than point an accusing finger at the statistical naïvety of a few parasitologists; one hopes for positive response to tutorial advice as well. In addition to the two major topics, comments are given about regression analysis for dose-response studies and analysis of related multiple variables.

Analysis of Variance↗