Chlamydia trachomatis PCR (Cobas Amplicor) in women: endocervical specimen transported in a specimen of urine versus endocervical and urethral specimens in 2-SP medium versus urine specimen only.

The sensitivity of Roche Cobas Amplicor Chlamydia trachomatis polymerase chain reaction (PCR) including the internal control (IC) programme to identify inhibition, was investigated on 3 different samples from women: (1) swab samples from the urethra and the cervix pooled in 2-SP transport medium, (2) swab sample from the cervix transported in a urine sample from the same patient, and (3) urine sample alone. Out of the 2412 patients, 193 (8.0%) were chlamydia positive and in 14 of these the results showed discrepancies between sampling methods. The sensitivity of PCR on urethra/cervix, urine/cervix and urine was 98.4% (190/193), 97.9% (189/193) and 93.3% (180/193) respectively. The higher sensitivity of PCR on urethra/cervix and urine/cervix as compared with urine alone was statistically significant. Without the IC, the sensitivity of PCR on urethra/ cervix, urine/cervix and urine would have been 95.9% (185/193), 94.8% (183/193) and 90.7% (175/193) respectively. Factors influencing the rate of inhibition were also studied.

Magic Lite Chlamydia immunoassay in urogenital samples and urine versus chlamydial culture.

Magic Lite Chlamydia assay (commercial test kit for the identification of Chlamydia trachomatis) was evaluated on urogenital samples and urine with chlamydial culture as the reference method. The sediment of the transportation buffer of specimens which were Magic Lite positive but culture negative or toxic was investigated for elementary bodies with fluorescein-labelled anti-chlamydial antibodies. The prevalence of chlamydial infection among the 577 men investigated was 13.7% as estimated by culture and direct immunofluorescence and 6.4% among the 173 women. In order to improve the sensitivity a cut-off value below that recommended by the manufacturer was used. The sensitivity of Magic Lite in male urethral specimens was then 60.8% and that in female urethral/cervical specimens 90.9%. The specificity was 99.6% and 100%, respectively. In urine specimens the sensitivity of Magic Lite was 63.3% (men) and 63.6% (women). The specificity was 99.4% and 100%, respectively. The sensitivity of Magic Lite on male urethral specimens was related to the number of inclusion bodies per well in culture and it was higher among men attending with clinical findings of urethritis (69%) than among asymptomatic men sampled as a screening procedure (36%) (P < 0.05). Corresponding differences between the sexes and between those with and without symptoms were not noted for Magic Lite applied on urine samples.

Endemic versus non-endemic gonorrhoea in Stockholm: results of contact tracing.

The aim of the present study was to characterize endemic versus non-endemic gonorrhoea to identify risk groups for transmission and to evaluate the effects of intensified contact tracing performed by specially trained social workers at venereal clinics. A total of 671 gonorrhoea patients (283 women and 388 men) comprised the study group. Seventy percent of the women and 48% of the men had an endemic infection (P < 0.001). Men without a steady partner had an increased risk of non-endemic infection. A decrease from 75% to 40% was noted in the proportion of endemic infection in Stockholm from the first quarter of the study period (2 years) to the last. Contact tracing resulted in 1.2 identified partners per patient. A total of 736 partners were examined either as a result of contact tracing efforts or because they had sought medical care on their own prior to intervention. Forty-seven percent of these partners were infected, 44% were not infected and 9% were examined outside the study with results unknown to us. The partner notification efforts yielded 161 new untreated cases. Contact tracing of women generated one new case per 4.0 interviewed women and contact tracing of men one new case per 4.3 interviewed men. Interviewing index patients with endemic infection yielded the highest number of new cases. Forty-three percent of the patients were infected outside Stockholm but only a smaller part of these patients spread their infection further into the community.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibiotic susceptibility, serovars and auxotypes of gonococcal isolates in Stockholm. Relation to geographical origin of the infection.

The antibiotic susceptibility, serovars and auxotypes were investigated in gonococcal strains isolated from all patients with gonorrhoea during one year in Stockholm, Sweden. The results were correlated to geographical origin of the infection. A total of 394 gonococcal strains were isolated from 392 patients, 135 (34%) women and 257 (66%) men. Beta-lactamase-producing gonococcal strains (PPNG) were isolated from 5% of the women and 16% of the men. Men had acquired their infection abroad more often than women (54% vs 33%) (P < 0.001). The majority (81%) of the PPNG infections were imported. Some serovars and auxotypes were more common among imported strains than among indigenous ones. All strains were sensitive to spectinomycin and 2 strains had decreased susceptibility to norfloxacin and ciprofloxacin. Decreased susceptibility to benzylpenicillin, ampicillin, doxycycline and cefuroxime was related to the geographical origin of the strains with strains imported from regions other than Europe being the most resistant.

Characterisation of PPNG and non-PPNG Neisseria gonorrhoeae isolates from Singapore.

OBJECTIVE: To characterise Neisseria gonorrhoeae isolates from Singapore. DESIGN: Characterisation of Neisseria gonorrhoeae isolates by auxotyping, serological analysis and plasmid profile analysis. SPECIMENS: Sixty randomly collected isolates from 41 symptomatic, untreated males and 19 female prostitutes were studied. RESULTS: Auxotyping of 25 PPNG and 35 non-PPNG strains showed that the Pro-auxotype was prevalent among both PPNG (56%) and non-PPNG (42.5%) strains. Prototrophic strains comprised 28% of PPNG and 32.5% of non-PPNG strains respectively. Serovar analysis showed that with the exception of seven serogroup WI strains, the majority belonged to serogroup WII/III. Serovar Aedih was predominant among both serogroup WI PPNG (80%) and non-PPNG (100%) strains. Serogroup WII/III PPNG strains were represented by nine serovars with the predominant serovars being Bacjk (28%) and Bcgjk (16%). Eleven serovars were identified in the WII/III non-PPNG strains and the major serovars were Bajk (20%), Bacjk (17%), Back (11.4%) and Beghjk (11.4%). Analysis of the 25 PPNG strains showed that 16 of them carried the 4.4 MDa (Asian type) resistance plasmid and nine strains harboured the 4.4 MDa plasmid in conjunction with the 24.5 MDa transfer plasmid. The cryptic plasmid of 2.6 MDa was present in 27 of the 35 non-PPNG strains. Five of the non-PPNG strains harbouring the cryptic plasmid also contained the 24.5 MDa transfer plasmid. The plasmid combination of 2.6 + 7.8 + 24.5 MDa was detected in three non-PPNG strains. CONCLUSION: The combination of epidemiological methods used in this study indicated the heterogeneity of N gonorrhoeae strains in Singapore. A total of 16 different combinations of auxotype, plasmid profile and serovar were seen in the 25 PPNG strains compared with 24 such combinations in the 35 non-PPNG strains. Such sensitive differentiation would otherwise not be possible using either auxotype-serovar (A/S) or auxotype-plasmid analysis.

Comparison of Pharmacia Chlamydia EIA, IDEIA and cell culture in the detection of urogenital chlamydial infection.

Two commercial test kits, Pharmacia Chlamydia EIA (PhEIA) and IDEIA Chlamydia Test, for the identification of Chlamydia trachomasis and McCoy cell culture were compared in urogenital specimens. The sediments of the transportation buffers of specimens with discordant results were investigated for elementary bodies (EB) with fluorescein-labelled antichlamydial antibodies. The prevalence of chlamydial infection among the men was 16% (48 of 293), 47 culture positive and one EB positive, and among the women 10% (10 of 97), 10 culture positive. In men, the sensitivity of PhEIA, IDEIA and culture was 71%, 40% and 98%, respectively. In women, irrespective of site, corresponding figures were 100%, 80% and 100%. The specificity and positive predictive values were 100% for both enzyme immunoassays in men and women. The low sensitivity of IDEIA could not be explained by the degree of infection as measured by the number of inclusion bodies in cell culture, the presence of antigen as measured by the number of EBs or the sampling order.

Screening for Chlamydia trachomatis genital infection in young men in Stockholm.

The purpose of this study was to investigate the prevalence of genital chlamydial infection in male military recruits in Stockholm, Sweden. Two hundred and thirty-eight men who admitted to sexual experience were included in the study. One hundred and five (44%) of these 238 men consented to a specimen being taken for chlamydial culture. Eleven (10%) of these 105 men had a positive chlamydial culture. All 11 men were 17-18 years old and 10 were asymptomatic. In order to decrease the prevalence of chlamydial infection, screening of young men is as essential as screening of women.

Sexually transmitted diseases in men in Mogadishu, Somalia.

The prevalence of viral and bacterial sexually transmitted diseases were studied in 101 men attending a dermatovenereal outpatient clinic in Mogadishu. A control group of 103 healthy adult men were included for the serological part of the study. Serological markers of hepatitis B virus (HBV), human immunodeficiency virus (HIV), cytomegalovirus (CMV) and herpes simplex virus (HSV) were studied. All sera were tested for syphilis markers. HBV serum markers were detected in 84% of the men in the study group and 66% of the healthy controls (P less than 0.005). Hepatitis B virus carriers were detected more frequently in the study group than among the controls. Also, 96% of the men in both groups had CMV antibodies and all of them had antibodies to HSV. No sera were found to contain HIV antibodies. The TPHA-positivity was 10% and 3% in the study and control groups respectively, and 5% of the patients had syphilis IgM antibodies. Sexual contact with prostitutes was recorded in 54% and 48% respectively of patients and controls, and such contact was correlated with TPHA-positivity in the study group. Chlamydia trachomatis antigen was detected in urogenital specimens of 14% of the men in the study group and gonococcal culture was positive in 53% of those with urethral discharge.

Rapid in situ generation of DNA restriction endonuclease patterns for Neisseria gonorrhoeae.

Restriction endonuclease (RE) digestion patterns of 26 isolates of Neisseria gonorrhoeae representing different serovars of serogroups WI, WII, and WIII were generated by agarose pellet entrapment and in situ digestion with HinfI and BglII. The method was fast, simple, and reproducible, and stable RE patterns were produced from subsequent in vitro passages. The cost of culture materials was reduced considerably, and no toxic or flammable solvents needed to be used. Excellent resolution of DNA fragments of higher molecular weights was obtained as a result of minimal mechanical shearing of the DNA. The REs HinfI and BglII were discriminative in the fragment length ranges of 2 to 6.5 and 2.5 to 21.5 kilobases, respectively. On the basis of densitometric scanning of electrophoretograms generated by HinfI digestion, the 26 isolates representing 12 serovars were divided into seven groups. BglII was found to be more discriminative; 15 RE patterns were established among the 26 isolates. Patterns generated by both REs showed that there was no correlation between a particular RE pattern and a serovar, since strains with identical RE patterns were from different serovars. With the exception of two strains (D3 and D14), which demonstrated positive correlation when both enzymes were used, all strains with identical serovar patterns had different RE patterns.

Genotypes and phenotypes of beta lactamase producing strains of Neisseria gonorrhoeae from African countries.

The phenotypes and genotypes of 26 beta lactamase (penicillinase) producing strains of Neisseria gonorrhoeae (PPNG) from African countries were investigated. Using the restriction enzyme technique nine different restriction enzyme patterns were found, two of them in 15 strains. Of the 26 strains, 16 belonged to serogroup WI (containing protein type IA) and 10 to serogroup WII/III (containing protein IB). Among the IA strains four different serovars were represented, whereas six serovars were found among the IB strains. Five different auxotypes were identified, of which proline requiring (found in 12 strains) and prototrophic (found in 10 strains) dominated. Twelve strains harboured a 4.4 megadalton as well as a 24.5 megadalton plasmid. A 3.2 megadalton plasmid was found in 14 strains, one of which also harboured a 24.5 megadalton plasmid. The 2.8 megadalton cryptic plasmid was present in all 26 strains. The MICs of doxycycline ranged from 0.25 to 2.0 mg/l; the MIC 50% for WI strains was 0.25 mg/l and for WII/WIII strains 1.0 mg/l. A total of 10 different combinations of restriction enzyme pattern, serovar, auxotype, and plasmid were seen in the 16 WI strains compared with eight such combinations in the 10 WII/WIII strains. As expected, the restriction enzyme technique and serological classification gave better differentiation than plasmid profiles and susceptibility to doxycycline. More relevantly, however, these techniques also compared favourably with auxotyping. When the different systems were combined, the sensitivity was greatly increased.

Serovars, auxotypes, and plasmid profiles of PPNG strains with Asian type plasmid isolated in Amsterdam.

In 1982 an increase of penicillinase producing strains of Neisseria gonorrhoeae (PPNG), carrying the 4.5 megadalton Asian type plasmid and the 24 megadalton transfer plasmid (Asia+), was observed in Amsterdam. The main auxotypes were proline requiring (Pro-) and proline and hypoxanthine requiring (Pro-Hyx-). Using two monoclonal antibody systems, it was shown that the serovars of strains with these auxotypes isolated in 1981 were different from those isolated in 1982, which indicated the start and end of microepidemics. Different serovars were also observed in Pro- and non-requiring (NR) Asia- PPNG strains isolated in 1981-2 and 1985 respectively. Only one serovar (Aedih/Arst) was common in strains isolated in 1981-2 as well as in 1985.

Comparison between serological classification and auxotyping in the analysis of Neisseria gonorrhoeae infections.

Auxotyping and serological classification, using monoclonal antibodies, were performed on 730 gonococcal strains. These strains were isolated from 725 consecutive patients seen at the Venereal Outpatients Clinic at the Department of Dermatology, Södersjukhuset, Stockholm, during one year, up to April 1983. Strains from patients with repeated gonococcal infections were, besides auxotyping and serological classification, analysed with restriction endonuclease cleavage. The strains were distributed into 16 auxotypes, of which the eight most common accounted for 97.4% of all isolates. The same strains were distributed into 38 serovars, of which the eight most common accounted for 88.4% of all isolates. When the two methods were combined, 98 combinations of auxotypes and serovars were seen. The eight most common combinations included 60.0% of all strains. Correlations were found between auxotypes and serogroups as well as serovars. The serological classification gave a better resolution compared with auxotyping; however, when the two systems were combined the sensitivity was highly increased.

Serovars, auxotypes, and plasmid contents of PPNG strains from outbreak in Amsterdam.

The first outbreak of penicillinase producing strains of Neisseria gonorrhoeae (PPNG) in Amsterdam in 1981-2 was caused mainly by African strains carrying the 24 megadalton transfer plasmid (Afr+) that were non-requiring (NR) and inhibited by phenylalanine (pheni), but African strains without the transfer plasmid (Afr-) that were NRpheni and Afr+ NR strains were also found. Serological classification, using two monoclonal antibody systems, showed that three main serovars (Ae/Av, Aedih/Arst, and Bacejk/Brpyust) could be distinguished in these PPNG strains, which indicated exchanges of plasmids in these serovars. The serovar Ae/Av predominated in the Afr+ and Bacejk/Brpyust in the Afr- strains.

Serological ecology of Neisseria gonorrhoeae (PPNG and non-PPNG) strains: Canadian perspective.

One hundred and thirty eight penicillinase producing Neisseria gonorrhoeae (PPNG) and 239 non-PPNG strains were characterised serologically using a panel of seven monoclonal antibodies directed against protein 1A and seven against protein 1B. An association between serovar and susceptibility to antimicrobial agents, auxotype, and plasmid content was observed. Serogroup WI strains were more sensitive to penicillin, ampicillin, tetracycline, erythromycin, cefoxitin, and cefuroxime. Sixty five (82%) of the 79 WI strains were typed as being serovar Aedgkih, and 47 (72%) of these strains required arginine, uracil, and hypoxanthine for growth (AUH-). Seventy one (44%) of 160 WII/WIII strains were serovar Bacejk, and 42 (59%) of these required proline, citrulline, and uracil for growth (PCU-) and were plasmid free. Serovars Bcgk, Beghjk, Bacjk, and Bajk were associated with resistance to antimicrobial agents. Analysis of PPNG isolates showed a new serovar, Af, which was associated with strains imported from Malaysia and Singapore that required proline and ornithine for growth (Pro-Orn-) and carried the 24.5 megadalton transfer plasmid, the 2.6 megadalton cryptic plasmid, and the 4.5 megadalton penicillinase producing plasmid. Other associations between serovar and geographical location were noted.

Serologic classification and contact-tracing in the control of microepidemics of beta-lactamase-producing Neisseria gonorrhoeae.

Serologic classification of isolated gonococcal strains and thorough contact-tracing were proved to be valuable in controlling an indigenous outbreak of infections caused by beta-lactamase-producing Neisseria gonorrhoeae (PPNG). Only 1-2% of gonococcal strains isolated in Sweden are PPNG, and before 1983 most of them were imported. During January-August 1983, 43 PPNG strains were isolated from 42 patients in Gothenburg. The PPNG strains were auxotyped and classified serologically. PPNG strains of serogroup WI, serovar (subgroup) Ae and of the prototrophic auxotype were isolated from 27 patients, including six prostitutes. Information obtained at interviews with these patients indicated that there had been a chain of infections. Quick and thorough contact-tracing stopped this microepidemic within three months. The serologic classification of the PPNG strains helped us to concentrate the epidemiologic efforts on those persons known to be infected with the epidemic strain, to trace this infection to other parts of Sweden, and to determine when the outbreak was finished.

Gonococcal serovar distribution in Stockholm, with special reference to multiple infections and infected partners.

Serological classification with co-agglutination, using monoclonal antibody reagents, was used to classify gonococcal strains from 731 consecutive patients seen at the Venereal Outpatients clinic at the Department of Dermatology, Södersjukhuset, Stockholm, up to April 1983. The strains could be divided into two serogroups, WI and WII/III. For the identification of strains belonging to serogroup WI, six Protein IA specific antibody reagents were used, and for strains of serogroup WII/III, seven Protein IB specific antibody reagents. The serogroup WI could be further subdivided into eight different serovariants (serovars), and serogroup WII/III into 30 different serovars. All strains reacted with at least one monoclonal antibody reagent and no strain reacted with both WI and WII/III specific reagents. In both serogroups there was one serovar that was common among women and heterosexual men and another which was more frequent among homosexual men. The 84 contact pairs had strains of corresponding serovar in 95%. Among 258 patients with two or more gonococcal isolates on the same occasion, the isolates from 93% had the corresponding serovar. Repeated gonococcal infections were more frequent among heterosexual men than among women and more frequent among homosexual than among heterosexual men. The serological classification of N. gonorrhoeae is a stable and rapid method and a useful epidemiological tool.

Use of gonococcal W serogrouping in the evaluation of a clinical trial of rosoxacin.

Serogrouping by co-agglutination was used for the characterization of isolates of Neisseria gonorrhoeae in a clinical trial of rosoxacin in Stockholm. Twenty-four isolates (56%) belonged to serogroup WI, 17 (40%) to WII, and two (5%) to WIII. The proportion of WI isolates in Stockholm was reported in earlier studies to be approximately 40%. On the basis of serogrouping data and clinical information, five (10%) of 48 patients in this study were classified as therapeutic failures. Of the initial WI isolates, 22 (92%) of 24 were inhibited by less than or equal to 0.03 microgram of rosoxacin/ml, as compared with ten (59%) of 17 of the initial WII isolates (.05 greater than P greater than .01). Thus, this study might underestimate the failure rate as compared with that for patient populations in which WII isolates are more prevalent, since WI isolates are more susceptible to rosoxacin than WII isolates. A certain WII serovar was correlated with decreased susceptibility to rosoxacin (P less than .001). Correlations were found between decreased susceptibility to rosoxacin and decreased susceptibility to other antibiotics (P less than or equal to .01). A high frequency of side effects (40%) was seen among the patients studied.

Susceptibility of Neisseria gonorrhoeae to rifampicin and thiamphenicol: correlation with protein I antigenic determinants.

Tests of susceptibility to thiamphenicol and rifampicin were done for 85 strains of Neisseria gonorrhoeae, including 28 beta-lactamase-producing and three spectinomycin-resistant isolates. Strains were serogrouped by co-agglutination with 14 monoclonal antibodies that are specific for different antigenic determinants on protein I of the gonococcal outer membrane. Thus the isolates could be classified into one of the serogroups WI, WII, or WIII and could be subgrouped further into several serovars. The minimal inhibitory concentration (MIC) of thiamphenicol ranged between 0.125 and 4.0 micrograms/ml and that of rifampicin between 0.016 and 4.0 micrograms/ml. All isolates with a MIC of rifampicin of greater than or equal to 0.5 microgram/ml had a MIC of thiamphenicol of greater than or equal to 2.0 micrograms/ml. Decreased susceptibility (thiamphenicol MIC, greater than or equal to 1.0 microgram/ml; rifampicin MIC, greater than or equal to 0.5 microgram/ml) to both drugs was correlated with serogroup WII specificity and also correlated with strains that belonged to serovars characterized by a positive reaction with the WII monoclonal antibody 2G2.

Antimicrobial susceptibility and susceptibility testing of Mycoplasma hominis: a review.

The determination of the minimal growth-inhibiting concentration (MIC), the minimal metabolism-inhibiting concentration (MMC), and the minimal mycoplasmacidal concentration (MCC) of various antimicrobial compounds for Mycoplasma hominis is influenced by the pH of the test media, the inoculum size, and the incubation time, although each of these factors generally do not affect the minimal concentration more than fourfold. M. hominis is resistant to beta-lactam antibiotics, vancomycin, sulfonamides, trimethoprim, and polymyxin B. There are great differences in the susceptibility of M. hominis to various macrolide antibiotics. Thus the organism is resistant to erythromycin and oleandomycin, moderately resistant to tylosin and spiramycin, susceptible to josamycin as well as to another macrolide drug, labelled M-4365G. M. hominis is also highly susceptible to the macrolide-like compound rosaramicin and to the tetracyclines (although resistant strains occur). It is susceptible to lincomycin and clindamycin, and moderately susceptible to chloramphenicol and rifampicin. The aminoglycosides have limited activity against M. hominis.