Accurate and efficient diagnostic techniques are an essential requirement of efforts to prevent and treat sexually transmitted diseases. Nucleic acid hybridization assays will be developed for this purpose using new technologies recently developed in our laboratory. These techniques include a chemical method for labeling probes with biotin and assay formats in which hybridization reactions take place in solution and in which assays can be performed in microtiter plates and the results can be measured spectrophotometrically. The format involves a reaction between DNA sequences and complementary DNA sequences. The probe, either DNA or RNA depending on the nucleic acid analyze to be detected, is labeled with biotin. Labeled hybrids are detected in an immunoreaction with a solid phase anti-biotin antibody and enzyme labeled monoclonal antibody specific for DNA-RNA hybrids. Monoclonal solution hybridization assays will be developed and standardized for the detection of Chlamydia trachomatis and human papillomaviruses. A prospective evaluation of the performance of these hybridization assays compared to other diagnostic systems for the detection of these microbes will be performed with urethral and cervical specimens from patients attending an STD clinic. To expand the diagnostic capabilities of hybridization assays, a genomic library from an LGV strain of C. trachomatis will be constructed and the library will be screened for genotype specific clones. Clinical isolates of C. trachomatis will be tested in hybridization assays using genotype probes in order to determine the correlation between genotype, classical serovar and biotype defined by clinical characteristics of the patient from whom the isolate was obtained. The successful development of practical techniques for detection of nucleic acids in clinical specimens may improve the care of patients with STDs and allow for greater understanding of the epidemiology and pathophysiology of these infections.

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Johns Hopkins University
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Thomas, D L; Zenilman, J M; Alter, H J et al. (1995) Sexual transmission of hepatitis C virus among patients attending sexually transmitted diseases clinics in Baltimore--an analysis of 309 sex partnerships. J Infect Dis 171:768-75
Thomas, D L; Rompalo, A M; Zenilman, J et al. (1994) Association of hepatitis C virus infection with false-positive tests for syphilis. J Infect Dis 170:1579-81
West, S; Munoz, B; Bobo, L et al. (1993) Nonocular Chlamydia infection and risk of ocular reinfection after mass treatment in a trachoma hyperendemic area. Invest Ophthalmol Vis Sci 34:3194-8
Kessis, T D; Slebos, R J; Han, S M et al. (1993) p53 gene mutations and MDM2 amplification are uncommon in primary carcinomas of the uterine cervix. Am J Pathol 143:1398-405
Kessis, T D; Slebos, R J; Nelson, W G et al. (1993) Human papillomavirus 16 E6 expression disrupts the p53-mediated cellular response to DNA damage. Proc Natl Acad Sci U S A 90:3988-92
Coutlee, F; Bobo, L; Abbass, H et al. (1992) Detection of HPV-16 in cell lines and cervical lavage specimens by a polymerase chain reaction-enzyme immunoassay assay. J Med Virol 37:22-9
Upchurch, D M; Ray, P; Reichart, C et al. (1992) Prevalence and patterns of condom use among patients attending a sexually transmitted disease clinic. Sex Transm Dis 19:175-80
Muller, M; Viscidi, R P; Sun, Y et al. (1992) Antibodies to HPV-16 E6 and E7 proteins as markers for HPV-16-associated invasive cervical cancer. Virology 187:508-14
Bonnez, W; Kashima, H K; Leventhal, B et al. (1992) Antibody response to human papillomavirus (HPV) type 11 in children with juvenile-onset recurrent respiratory papillomatosis (RRP). Virology 188:384-7
Shah, K V; Daniel, R W; Simons, J W et al. (1992) Investigation of colon cancers for human papillomavirus genomic sequences by polymerase chain reaction. J Surg Oncol 51:5-7

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