An ELISA kit with two detection modes for the diagnosis of lymphatic filariasis.

The aim of this study was to develop a low-cost antifilarial immunoglobulin (Ig) G4 detection kit for the diagnosis of lymphatic filariasis. The kit was designed to be used by minimally trained personnel without the constraints of expensive laboratory equipment. We provide a description of the development and validation of a single-serum-dilution based enzyme-linked immunosorbent assay (ELISA) kit with ready-to-use reagents for measuring antifilarial IgG4 antibodies. The kit was tested on residents in Brugia malayi-endemic areas in southern Thailand. Detection was performed by naked-eye observation of the resultant colour of the immunological reactivity. The coefficient of variation (CV) was used to assess the reproducibility of the results. Long-term stability was measured over a 6-month period. Sensitivity of the test kit was 97% when compared with microfilariae detection in thick blood smears. Specificity was 98.7% based on the sera of 57 patients living outside the endemic areas who were infected with other parasites and 100 parasite-free subjects. All positive CVs were < 10%. The test kit was remarkably stable over 6 months. Field validation was performed by the detection of antifilarial IgG4 in 4365 serum samples collected from residents of brugian filariasis-endemic areas and compared with outcome colours of the test samples by the naked eye. Subsequent ELISA evaluation of these results using an ELISA reader indicated high agreement by the kappa statistic. These results demonstrate that the test kit is efficient and useful for public health laboratories as an alternative tool for the diagnosis of lymphatic filarial infection.

A cohort study on anti-filarial IgG4 and its assessment in good and uncertain MDA-compliant subjects in brugian filariasis endemic areas in southern Thailand.

The prevalence of Brugia malayi was surveyed in three highly endemic provinces in southern Thailand as part of an ongoing evaluation of the effectiveness of two rounds of mass drug administration (MDA). Prior to MDA IV, and thereafter every 6 months up to 18 months, including MDA V at 12 months, nocturnal blood sampling was assayed for microfilaria (mf) and IgG4 antibodies in 200 subjects from Nakorn Srithammarat and Surathani provinces. From an additional 300 subjects from Narathiwatt province, daytime blood was also examined for IgG4 antibodies for 1 year. Subjects who tested positive, with reciprocal IgG4 titres above 100, were dichotomized in the 'good compliant' or 'uncertain compliant' groups. Stool samples were examined for intestinal parasites from all 500 subjects simultaneously. Only two subjects (1%) tested positive for mf, while 98 subjects (19.6% of 500) tested positive for IgG4. In general, the good drug-compliant subjects were less likely to exhibit the IgG4 response than subjects in the uncertain compliant group. Only six subjects (1.2%) exhibited lymphoedema while 68 subjects (13.6%) had a total of 105 parasitic infections, with female instances of protozoan infections exceeding male instances of helminth infections. It was concluded that the two MDA rounds were highly successful in not only in reducing mf to negligible levels but also in lowering antifilarial IgG4 titres in the good compliant subjects. The IgG4 assay is a sensitive and cost-effective surveillance tool for the early detection of brugian infections that is not contingent on nocturnal blood collections.

Diagnostic value of IgG isotype responses against Brugia malayi antifilarial antibodies in the clinical spectrum of brugian filariasis.

To study the diagnostic significance of antifilarial IgG subclasses in the clinical spectrum of brugian filariasis, IgG1, IgG3 and IgG4 antifilarial antibodies were determined in an exposed population comprising 74 asymptomatic amicrofilaraemics, 30 microfilaraemics, 20 lymphangitis and 16 elephantiasis patients resident in Narathiwart province, an area endemic for Brugia malayi lymphatic filariasis in southern Thailand. The dominant isotype of antifilarial antibody was IgG4. A significantly higher percentage of individuals were positive for IgG1 in the microfilaraemic and lymphangitis groups compared with the elephantiasis and endemic normal patients, while a significantly higher positive rate of IgG3 was found in those with lymphangitis. The possible role of these isotypes for diagnostic purposes and the pattern of antibody response in various clinically manifesting groups are discussed.