Current methodologies for diagnosing infection with Neisseria gonorrhoea require specimens that are obtained by invasive means: urethral swabs from men and cervical swabs from women. These specimens are usually cultured onto Thayer-Martin media; alternatively a variety of other methods can be employed to diagnose gonorrhea, but none is as sensitive as culture. The objective of this proposal is to create an inexpensive, rapid, simple and non-invasive immunologic test for gonorrhea. It must be as sensitive as culture and applicable for use in developing countries. The project will be accomplished through 3 specific aims: (l) to optimize an already existing prototype assay called the HygEIA GC Test to use with non- invasive specimens, and to convert this ELISA format to an immunochromatographic (ICT) format with a membrane read-out. Initially, the assay will be developed using two existing antibodies, one directed against the H8 antigen (a current component of the HygEIA GC Test) and a gonococcal anti-lipooligosaccliaride antibody that reacts broadly with N. gonorrhoea; (2) to create polyclonal antibody libraries using a molecular cloning system that produces antibodies that recognize multiple epitopes on gonococcal surfaces exclusively; simultaneously these libraries will be immortalized and can be utilized time and again to produce, antibodies with reproducible specificities and; (3) to use patients at risk for gonorrhea to test prototype formats. Through overlapping strategies, beginning with a prototype that already functions, it is anticipated that a final product will emerge.
|Sharon, J; Sarantopoulos, S; Den, W et al. (2000) Recombinant polyclonal antibody libraries. Comb Chem High Throughput Screen 3:185-96|