Identification of Recombinant Antigens of Mycobacterium Leprae Which React With Antibodies from Lepromatous and Borderline Leprosy Patients

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1991-06

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Addis Ababa University

Abstract

Recombinant antigens of Mycobacterium leprae were identified by screening a lambda gt-11 M. leprae library with absorbed lepromatous (LL), borderline tuberculoid (BT) leprosy and a healthy contact sera in separate experiments. Six, three, and one clones were respectively isolated by successive screenings of the plaques by immunodetection of antigens on nitrocellulose filters. Of these, a total of six lysogens were generated. Four were made from the clones isolated using LL serum and two were made fromclo~es isolated using BT serum. Analysis of the apparent molecular mass of the recombinant proteins by either SDS-PAGE or Western blotting, showed the fusion proteins to be in the range of 116 kD to 1.44 kD. The reactivity of recombinant proteins with patients sera throughout the spectrum of the disease was found to be heterogenous regardless of the molecular sizes of the proteins. The characterization of the clones with regard to DNA insert mass was made by amplification with Taq polymerase chain reaction. The analysis on 1.5% agarose gel electrophoresis showed the range to be from 0.3 Kb to 2.0 Kb. The proliferation of T-cell as a result of stimulation by the recombinant proteins (antigens) looks promising and further study for confirmation is in progress.

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Biology

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