Wuchereria bancrofti recombinant antigen-derived poly- and monoclonal antibodies for the detection of circulating antigen(s) in the sera of lymphatic filarial patients.

A sandwich antibody ELISA was employed for the detection of circulating filarial antigen in patients with bancroftian filariasis. Wuchereria bancrofti recombinant antigen-derived polyclonal and monoclonal antibodies were successfully used as the revealing antibodies and their efficiency was compared. All the microfilariae (mf) positive (by finger prick and examination of 20 microliters of blood under the microscope) individuals tested showed the presence of circulating antigen(s). Among the antigen positive endemic normals (mf negative by the finger prick method), 43% showed microfilariae by a sensitive parasitological method viz. membrane filtration of the night blood samples. A significant correlation was observed between the parasite antigen levels and the blood microfilaria counts among the mf carriers. This information on the parasite antigen levels could be an ideal monitor to indicate the degree of active infection and in the follow up of chemotherapy.

White-light imaging by use of a multiple passband acousto-optic tunable filter.

White-light imaging was accomplished by operation of a TeO(2) acousto-optic tunable filter (AOTF) with 40 simultaneous overlapping passbands from 400 to 700 nm. The AOTF was chromatically compensated by a wedge applied to the output surface of the AOTF, and the measured spatial resolution correlated well with predictions. Switching off specific rf's applied to the AOTF produced optical rejection corresponding to the inactive passbands. A rejection ratio of 30 dB was demonstrated, and the rejection level was found to be controlled by leakage through the sidelobes of adjacent passbands.

Cloning, over-expression and evaluation of a recombinant fusion protein of Wuchereria bancrofti towards its application as a diagnostic agent for bancroftian filariasis.

A low molecular weight (15 kDa) surface antigen of the cattle filarial nematode, Setaria digitata, was earlier shown to be specifically recognized by the antibodies from human bancroftian filarial (Mf positive) patients' sera (Theodore & Kaliraj, 1990). The filarial specific antibodies bound to a 15 kDa peptide in preparative Western blots were eluted and employed in screening of candidate antigens expressed in the genomic library of Wuchereria bancrofti at the IgG4 subclass antibody level. A recombinant clone (lambda WbG7) reacting strongly with filarial sera but poorly with sera from patients infected with non-filarial helminths was selected for further studies. The 2 kb DNA insert of the clone lambda WbG7 was recloned into a pMAL vector and the recombinant clone pGT7 thus obtained was over-expressed and affinity purified. The purified 105 kDa fusion protein of the clone pGT7 was specific and was not recognized by the non-filarial sera at the IgG4 level. All microfilaraemic individuals were positive by IgG4 assay. However, similar attempts to diagnose by filarial-specific IgE assays failed to recognize microfilaraemic individuals. Moreover, by filarial-specific IgG4 assays, the endemic normals were distinctly divided into two groups, showing higher and lower recognition for this antigen indicating current and past/no infection. Among the filarial-IgG4 (assay)-positive 'endemic normals', 14% showed 'microfilariae' during repeated peripheral night blood examination, confirming the validity of the recombinant antigen, pGT7 based assay.

Isolation, purification and characterization of surface antigens of the bovine filarial parasite Setaria digitata for the immunodiagnosis of bancroftian filariasis.

The surface antigens of the bovine filarial parasite Setaria digitata were isolated by EDTA extraction and purified by affinity chromatography using sepharose bound human filarial (Wuchereria bancrofti) antibodies obtained from chronic human filarial sera. The purified and crude antigens were used in enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibodies in bancroftian filariasis. The purified antigen showed sensitive and specific reactions in ELISA for the detection of antibodies in filarial sera and showed least cross reactivity with other parasitic infections. The crude and purified antigens showed about 18 and 6 peptide bands respectively in SDS-PAGE and about 11 and 6 antigenic bands respectively in enzyme-linked immunoelectrotransfer blot (EITB). The purified antigen was observed to be glycoprotein in nature. It was possible to identify the stage-specific infection in human filariasis by using the crude and purified antigens in EITB.