In a controlled trial of rosoxacin in patients with non-gonococcal urethritis (NGU), 150 mg of the antibiotic given twice daily for 10 days was compared with 300 mg triple tetracycline (Deteclo) given twice daily for the same period. Only six (19%) of 31 patients treated with rosoxacin were free of urethritis after 10 days; Chlamydia trachomatis was reisolated from 12 (92%) of 13 patients who were chlamydia positive originally, and Ureaplasma urealyticum was reisolated from 12 (80%) of 15 patients who were ureaplasma positive originally. In contrast, 18 (58%) of 31 patients treated with triple tetracycline were cured clinically after 10 days; C trachomatis was not reisolated from any of 10 patients who were chlamydia positive originally, and U urealyticum was reisolated from only three (17%) of 18 patients who were ureaplasma positive originally. These results were consistent with the antimicrobial inactivity of rosoxacin in vitro and they cannot be reconciled with previous reports of successful use of this antibiotic in NGU. Ureaplasmas were isolated more frequently and in larger numbers from chlamydia negative than from chlamydia positive patients, but it is probable that ureaplasmas resistant to tetracycline were not responsible for persistent urethritis.

The chlamydial genus-specific antigen was extracted with phenol/chloroform/petroleum ether (PCP) from preparations of Chlamydia trachomatis and C. psittaci, and quantities measured using an assay for lipopolysaccharide (LPS). The LPS from C. trachomatis contained 2.2% (w/w) of ketodeoxyoctanoic acid. Five IgG monoclonal antibodies reacted in an ELISA with LPS from both species, the antigen being periodate-sensitive and heat-resistant, confirming that all antibodies were against the genus-specific antigen. All the antibodies bound to the PCP extract of C. trachomatis on an immunoblot, at a position corresponding to the periodate-Schiff-stained bands of both C. trachomatis extract and Salmonella Re-LPS. When linked to trypsin-treated sheep erthrocytes and used in reverse passive haemagglutination tests, all antibodies gave indicator cells capable of detecting chlamydial LPS or crude preparations of chlamydiae grown in McCoy cells, the sensitivity varying with the antibody used. The antibodies varied in IgG subclass (either IgG2a or IgG3), and in ability to precipitate in immunodiffusion tests. Two antibodies cross-reacted with one strain of Acinetobacter in ELISA and with Salmonella Re-LPS in both ELISA and immunodiffusion tests. The other three did not react in ELISA with Acinetobacter strains or Salmonella Re-LPS, and none of the five reacted with LPS of E. coli or Pseudomonas morsprunorum.

Journal Article DETECTION OF CHLAMYDIAE BY ISOLATION AND DIRECT EXAMINATION Get access R T EVANS, MIBiol, R T EVANS, MIBiol Division of Communicable Diseases Clinical Research CentreHarrow Search for other works by this author on: Oxford Academic PubMed Google Scholar R M WOODLAND, PhD R M WOODLAND, PhD Sub-Department of Virology Institute of OphthalmologyLondon Search for other works by this author on: Oxford Academic PubMed Google Scholar British Medical Bulletin, Volume 39, Issue 2, 1983, Pages 181–186, https://doi.org/10.1093/oxfordjournals.bmb.a071813 Published: 01 June 1983

Chlamydial group antigen was extracted from Chlamydia trachomatis strain SA2(f) and used as the antigen for an ELISA. The assay was reproducible since chlamydial antibody titres differed by no more than twofold when sera were tested on up to eight occasions. In tests on sera from 75 patients attending venereal disease or rheumatology clinics, the results of the ELISA and of a microimmunofluorescence (MIF) technique were similar for 61 of the sera, that is an 81% agreement. However, the ELISA was a little more sensitive than the MIF technique and at least tenfold more sensitive than the complement fixation procedure. Chlamydial IgG antibody at a titre of 1/greater than or equal to 16 was detected by the ELISA in 6% of children's sera, in 20% of sera from adult patients attending hospital with non-venereal diseases and in 85% of sera from persons attending venereal disease or rheumatology clinics. IgM and IgG antibodies were detected also by the ELISA in the sera of chimpanzees and marmosets which had been infected genitally with C trachomatis and, in general, the titres were greater than those recorded by the MIF test. The value of the ELISA in comparison with the MIF test is discussed.

McCoy cell cultures were inoculated with 121 urethral and cervical specimens taken from patients attending one of two sexually transmitted disease clinics. The mean number of Chlamydia trachomatis inclusions was greater when the cultures were inoculated with the specimens and centrifuged in the clinic than when the specimens were first stored in liquid nitrogen. Furthermore, 18 of the 29 chlamydia-positive specimens produced larger numbers of of inclusions when inoculated immediately. Despite this, the isolation rate from specimens inoculated directly (22%) was about the same as from specimens which had been frozen (21%). Of the 30 occasions on which the cell monolayers were disrupted, 29 followed immediate inoculation. This may possibly have been due to some toxic factor in some of the specimens.

A laboratory-passaged genital strain of Chlamydia trachomatis and two unpassaged genital strains from patients with nongonococcal urethritis were inoculated intraurethrally into three young male chimpanzees. Chlamydia were recovered from the urethra of two animals and specific antibody responses were detected in all of them. Furthermore, a urethral polymorphonuclear leucocyte response, but not an overt discharge, occurred in all the chimpanzees about 1-2 weeks after inoculation. None of these events occurred in a chimpanzee inoculated with medium only. At necropsy three months after inoculation the submucosa of the urethra of one chimpanzee was densely infiltrated with small round cells. This suggests that a similar chronic lymphocytic response may occur in human chlamydial infection of the urethra.

SummaryFollicular cervicitis was recognised in 15 (44 per cent) of 34 women who were examined colposcopically and who were sexual partners of men with non‐gonococcal urethritis. Valid results of culture for Chlamydia trachomatis were obtained in 26 cases: the organism was isolated from the cervix of five of 11 women in whom follicular cervicitis had been diagnosed, but from only one of 15 whose cervices did not have this change. A similar correlation was not found for infection with Mycoplasma hominis or Ureaplasma urealyticum.

The 24-hour delay between seeding coverslips with cells and inoculating samples for culture of chlamydiae was reduced to less than 1 hour by using coverslips which had been pre-treated with glutaraldehyde-activated gamma-aminopropyl-triethoxysilane. Treated coverslips were not toxic for McCoy cells and even one year after treatment monolayers formed rapidly on them. Furthermore, all of 13 Chlamydia trachomatis serotypes and one C. psittaci strain tested produced inclusions in such cell monolayers. In comparative tests, when there were large numbers of inclusions, more were always seen in conventionally produced monolayers than in monolayers on treated coverslips. However, when there were few inclusions, more were seen in the latter monolayers, a phenomenon observed with unpassaged chlamydiae in clinical specimens as well as in laboratory-passaged strains. The rapid method is, therefore, as sensitive for isolating chlamydiae as using conventionally produced monolayers.

McCoy cells treated in six different ways, in addition to untreated cells, were compared to determine which gave rise to the largest number of Chlamydia trachomatis inclusions when tested with a laboratory-passaged strain. The same batch of cells was treated by irradiation, preinoculation exposure to cytochalasin B or 5-iodo-2'-deoxyuridine, and postinoculation exposure to cycloheximide, hydrocortisone, or emetine. Significantly more inclusions were always found in cells which had been treated with cycloheximide than in cells treated in any other way. Conversely, untreated McCoy cells always had significantly fewer inclusions than cells which had received some form of treatment. Similar results were obtained when cycloheximide-treated, irradiated, and untreated cells were inoculated with urethral specimens containing unpassaged organisms.

Journal Article The incidence of tetracycline-resistant strains of Ureaplasma urealyticum Get access R. T. Evans, R. T. Evans MRC Clinical Research Centre, Division of Communicable DiseasesWatford Road, Harrow, Middlesex HA1 3UJ, England Search for other works by this author on: Oxford Academic PubMed Google Scholar D. Taylor-Robinson D. Taylor-Robinson MRC Clinical Research Centre, Division of Communicable DiseasesWatford Road, Harrow, Middlesex HA1 3UJ, England Search for other works by this author on: Oxford Academic PubMed Google Scholar Journal of Antimicrobial Chemotherapy, Volume 4, Issue 1, January 1978, Pages 57–63, https://doi.org/10.1093/jac/4.1.57 Published: 01 January 1978 Article history Accepted: 12 August 1977 Published: 01 January 1978