APPROACHING A METHOD TO DETECT THE ETIOLOGIC AGENTS OF INTERSTITIAL CYSTITIS
A Thesis
Submitted to the Graduate Faculty of the Medical Center of
Louisiana State University and
Agricultural and Mechanical College
in partial fulfillment of the
requirements for the degree of
Master of Science
in
The Department of Microbiology, Immunology and Parasitology
by
Kuoting Wu
M.S., Fu Jen Catholic University, 1990
April, 1996
Abstract
Interstitial cystitis (IC) is a chronic inflammation of the bladder of unknown etiology, which is characterized by bladder pain, urinary frequency and urgency. The purpose of this work was to develop a method to identify potential infectious agents in IC patients’ bladders. In collaborative work, these extracted DNAs from IC patients’ bladder biopsies were polymerase chain reaction (PCR)-amplified by bacterial 16S rRNA universal primers; PCR products were cloned into pUC18. In this thesis, the clones were sequenced and analyzed. To sequence these clones (pUC18-IC) efficiently, inserted DNA was PCR amplified by either forward or reverse biotinylated primers. Streptavidin-bound magnetic beads were used to separate biotinylated single-stranded DNA and dideoxy DNA sequencing was performed on this bead-bound single-stranded DNA. This novel method may be useful to design experimental treatments of IC patients and aid in the development of a convenient diagnostic method for clinical laboratories. By two computer database (RDP and NCBI) search of bacteria sequences, we obtained different results; however, similarity to possible pathogens, E. coli and Pseudomonas spp. was consistently observed. Nevertheless, the definitive etiologic agents of IC are still unknown. In the future, identification of the true agent of IC may require animal models and immunological studies.