Evaluation of the indirect haemagglutination test in the diagnosis of amoebiasis

The IHA test was evaluated in the diagnosis of amoebiasis. Axenically-grown E. histolytica was used as an antigen. A total of 427 sera from symptomatic (intestinal and extra-intestinal) amoebiasis patients. asymptomatic carriers, patients with parasitic intestinal infections other than E. histolytica, and healthy controls were tested. From 288 symptomatic cases of amoebiasis, 232 (80.6 per cent) gave positive reactions. In 93 asymptomatic cases of amoebiasis, 55 (59.1 percent) were seropositive. In testing of sera from 16 subjects with parasitic intestinal infections other than E. histolytica, a low-level positive IHA titres occurred in 2 (12.5 per cent). The test was also positive with a low titre in 3 (10.0 per cent) of the 30 subjects of the healthy control group. The results indicate that the IHA test is of value in the confirmation of intestinal and extra-intestinal amoebic infections especially in cases where the parasite is difficult to demonstrate.     

The serodiagnosis of human hydatid disease: 2. Additional studies on selected sera using indirect haemagglutination (IHA), enzyme linked immunosorbent assay (ELISA) and defined antigen substrate spheres (DASS).

Sixty-one serum samples selected on the basis of reactivity in the complement fixation (CF) and latex agglutination (LA) test, were further examined for sensitivity and specificity by indirect haemagglutination (IHA), enzyme linked immunosorbent assay (ELISA) and defined antigen substrate spheres (DASS). Twenty sera from healthy Europeans and 48 samples from patients with either schistosomiasis or trichinosis were also tested. Comparable levels of sensitivity were found between the CF and LA positive sera and IHA, ELISA and DASS. Of the CF positive LA negative group of sera, many were positive by DASS but only a few reacted in IHA and ELISA. Some cross reactivity was also observed in the schistosomiasis sera tested by IHA and ELISA.     

Indirect haemagglutination reaction (LH)-the method of choice for the detection of anamnestic antibodies to varicella - zoster (VZ) virus.

The method of indirect haemagglutination is a sensitive quantitative serological reaction for the detection of anamnestic titres of VZ antibodies. The curve obtained by the examination of a representative population sample has shown that the occurrence of VZ antibodies increases with the age so that 60% of children of under-school age and 90% of 15-year-old children have antibodies to varicella-zoster virus. In adult age groups, seropositivity ranges between 90--100%. The incidence and level of antibodies are identical in both sexes and do not decrease in old age groups. The mean titre of indirect haemagglutinating antibodies was 1 : 128.     

Preparation of standard amoeba-antigen from axenicEntamoeba histolytica and its use in the serodiagnosis and seroepidemiology of amoebiasis

A method described for large scale cultivation ofEntamoeba histolytica axenically in a modified Diamond’s TP-S-1 monophasic medium. Crude amoebaantigen prepared by the ultrasonication of the trophozoites ofE. histolytica, was fractionated by sephadex G-200 column into four different fractions. The whole antigen and its different fractions were freeze-dried and upon reconstitution contained approximately 1.8 mg N/ml or roughly the equivalent of 10 × 10⁶ amoebae per ml. Both whole antigen and its fractions have been used for the detection of specific antibody in the patients’ sera. Rabbits were immunised with the antigen and the immunoglobulins were separated from hyperimmune sera by DEAE-cellulose chromatography and salt fractionations. Sera collected from different categories of amoebiasis patients, amoebic liver abscess, amoebic hepatitis, amoebic dysentery, and asymptomatic amoebiasis, were tested serologically using standard amoeba-antigen for serodiagnosis and epidemiological assay of amoebiasis. Results of the assay showed that standard amoeba-antigen is very useful for diagnosis of invasive amoebiasis.     

Comparison of the Kato-Katz technique, hatching test and indirect hemagglutination assay (IHA) for the diagnosis of Schistosoma japonicum infection in China

The Kato-Katz technique (duplicate 41.7 mg fecal smears), hatching test and indirect hemagglutination assay (IHA) were compared for their ability to detect human Schistosoma japonicum infection in two endemic villages (Zhonjiang and Zhuxi) in rural China. The hatching test (using a nylon bag, and based on about 30 g of feces) and IHA are conventional Chinese diagnostic methods. In both villages, the trends of prevalences with age and sex were comparable for the different methods. In Zhuxi, Kato-Katz examinations of stools from 7 different days and hatching were available, which could be used as a reliable gold standard. This resulted for IHA in a sensitivity of 80% and a specificity of 48%. The sensitivity of the Kato-Katz technique using one stool specimen was 68%, twice that of hatching (33%). In Zhonjiang, however, hatching resulted in more positive cases than Kato-Katz (prevalence 31% vs. 24%). Apparently, the result of the hatching test depends on environmental factors such as temperature and water quality. Although imperfect, Kato-Katz is recommended out of the three evaluated techniques as the method of choice for large-scale screening of S. japonicum. Hatching is much more tedious, provides inconsistent and only qualitative results, and is not much more sensitive than Kato-Katz. Its poor specificity makes IHA unsuitable for individual screening, but it may be more effective for community diagnosis.     

Development of passive haemagglutination (PHA) and haemagglutination inhibition (HAI) technique for potency estimation of Cobra Antisnake Venom Serum (ASVS).

Antibodies against snake venom or antivenom potency are assayed quantitatively by in-vivo neutralization test in mice, which requires large number of laboratory animals. In potency assays of biological substances such as antivenoms, it is highly desirable to avoid suffering and death of animals by substituting in-vivo method with in-vitro methods, provided such methods measure life-saving capability with precision similar to that of in-vivo method. The in-vitro tests determine the neutralizing power of antivenom by permitting the evaluation of a particular biological activity of the venom and its neutralization after mixing the venom with the antivenom [Theakston RDG, Reid HA. Development of simple standard assay procedures for the characterization of snake venom. Bull WHO 1983;61:949-956; Gutierrez JM, Rojas G, Lomonte B, Gene JA, Chaves F, Alvarado J, et al. Standardizing of assays for testing the neutralizing ability of antivenoms. Toxicon 1990;28:1127-1129; Theakston R.D.G. Comments on letter of Gutierrez et al. on standardization of assays for testing the neutralizing ability of antivenoms. Toxicon 1990;28:1131-1132; Harvey AL, Barfaraz A, Thomson E, Faiz A, Preston S, Harris JB. Screening of snake venom for neurotoxic and myotoxic effects using simple in-vitro preparation from rodents and chicks. Toxicon 1994;32:257-265; World Health Organization Progress in characterization of venom and standardization of anti-venoms. Geneva: WHO offset publication; 1981. p. 58.]. Hence, the ideal requirements for an assay in detecting venom and venom antibody include high level of sensitivity, specificity (ability to differentiate between venom and venom antibody produced by closely related species of snakes), reproducibility and simplicity. A new in-vitro procedure for quantitative analysis of potency of ASVS by passive haemagglutination (PHA) and haemagglutination inhibition (HAI) has been explored. The methods described are simple, rapid, economical, reproducible and useful in replacing the more expensive in-vivo neutralization assays. Moreover, it also eliminates the use of laboratory animals.     

Parasiticidal effect of 16α-bromoepiandrosterone (EpiBr) in amoebiasis and cysticercosis

The effect of the dehydroepiandrosterone analog 16α-bromoepiandrosterone (EpiBr) was tested on the tapeworm Taenia crassiceps and the protist Entamoeba histolytica, both in vivo and in vitro. Administration of EpiBr prior to infection with cysticerci in mice reduced the parasite load by 50% compared with controls. EpiBr treatment induced 20% reduction on the development of amoebic liver abscesses in hamsters. In vitro treatment of T. crassiceps and E. histolytica cultures with EpiBr, reduced reproduction, motility and viability in a dose- and time-dependent fashion. These results leave open the possibility of assessing the potential of this hormonal analog as a possible anti-parasite drug, including cysticercosis and amoebiasis.     

A comparison of the indirect haemagglutination test with the toxin neutralization test for the estimation of diphtheria antitoxin in mouse sera

Serum samples from 42 groups of mice immunized for different immunization periods with various doses of Adsorbed Diphtheria-Tetanus Vaccine, Adsorbed Diphtheria-Tetanus and Pertussis Vaccine and a standard diphtheria toxoid were assayed for their diphtheria antitoxin content by indirect haemagglutination (IHA) and by toxin neutralization (TN) tests. A very good correlation of 0.91 was obtained between the results of the two methods. There was no statistically significant difference between the IHA and the TN titres obtained. Adsorption with sheep red cells and treatment of the sera with 2-mercaptoethanol had no effect on the IHA titres. The minimum level of antitoxin detectable by the IHA test was 0.00039 IU ml-1. IHA proved to be a sensitive, specific and reproducible method which can be used reliably for the assay of diphtheria antitoxin in mouse sera.