Iona M. C. Martin

Rapid Sequence-Based Identification of Gonococcal Transmission Clusters in a Large Metropolitan Area

In large metropolitan areas, which typically have the highest rates of gonorrhea, the identification of chains of transmission by use of partner notification is problematic, and there is an increasing interest in applying molecular approaches, which would require new discriminatory high-throughput procedures for recognizing clusters of indistinguishable gonococci, procedures that identify local chains of transmission. Sequencing of internal fragments of 2 highly polymorphic loci, from 436 isolates recovered in London during a 3-month period, identified clusters of antibiotic-resistant and antibiotic-susceptible isolates with indistinguishable genotypes, the vast majority of which were also identical or closely related by other methods, and defined groups of individuals who typically had similar demographic characteristics. This discriminatory sequence-based approach produces unambiguous data that easily can be compared via the Internet and appears to be suitable for the identification of linked cases of gonorrhea and the timely identification of transmission of antibiotic-resistant strains, even within large cities.

Lymphogranuloma venereum in the United kingdom.

BACKGROUND Over the past 2 years, lymphogranuloma venereum (LGV), caused by L serovars of Chlamydia trachomatis, has emerged as a significant problem among men who have sex with men (MSM). We report on, to our knowledge, the largest case series of LGV to date, with detailed epidemiological and clinical characteristics of the epidemic in the United Kingdom. METHODS A national diagnostic service and surveillance system was established in October 2004. Cases were confirmed by the presence of C. trachomatis and an LGV serovar (L1, L2, or L3) from genotyping. For confirmed cases, an enhanced surveillance questionnaire was sent to the clinician. RESULTS Through February 2006, a total of 327 cases of LGV were confirmed. Cases were diagnosed across the United Kingdom, with the majority from London (71%) and Brighton (13%). Case reports were received for 282 MSM. The majority (96%) had proctitis, many with severe local and systemic symptoms. There was a high level of coinfection with human immunodeficiency virus (76%), hepatitis C (19%), and other sexually transmitted infections (39%). Nine cases of human immunodeficiency virus infection were diagnosed around the same time as LGV. Most cases were acquired within the United Kingdom, although patients with early cases were more likely to report contacts in The Netherlands. CONCLUSIONS We found a significant burden of this once-rare sexually transmitted infection among MSM in the United Kingdom. LGV may be contributing to the epidemic of human immunodeficiency virus infection by facilitating transmission. Further control efforts are required, including awareness campaigns, continued detailed surveillance, and expanded chlamydia testing among MSM.

Ciprofloxacin resistance in Neisseria gonorrhoeae in England and Wales in 2002

The Gonococcal Resistance to Antimicrobials Surveillance Programme (GRASP) monitors trends in antimicrobial resistance in consecutive gonococcal isolates from 26 genitourinary medicine clinics in England and Wales. In 2002, 2204 gonococcal isolates were tested, and the overall prevalence of ciprofloxacin resistance (minimum inhibitory concentration > or =1 mg/L) was 9.8%, compared with 3.1% in 2001 and 2.1% in 2000. Between 2001 and 2002, prevalence of ciprofloxacin resistance increased two to three-fold, irrespective of recent sexual contact overseas, sex, or residence within or outside of London. These findings suggest that national and local treatment guidelines need to be reviewed urgently.

Molecular Typing of Neisseria gonorrhoeae Causing Repeated Infections: Evolution of Porin during Passage within a Community

Thirty-three Neisseria gonorrhoeae isolates from 15 persons infected multiple times with the same serovar were compared using por gene sequencing, opa-typing, and arbitrarily primed-polymerase chain reaction. All three molecular techniques were more discriminatory than serotyping and identified differences between some isolates belonging to the same serovar. Although there were differences among Por sequences within some serovars, 10 of 15 subjects became reinfected with gonococci expressing identical Por proteins. Sequence analysis of por genes revealed evidence of horizontal genetic exchange and point mutations in potential surface-exposed regions during passage in the community.

A prospective social and molecular investigation of gonococcal transmission

BACKGROUND Gonorrhoea is a common infectious disease, poorly controlled despite effective treatments. Tracing chains of transmission is difficult, because sexual partners are commonly difficult or impossible to identify. We assess the use of gonococcal opa-typing in identifying transmission links not revealed through interview. METHODS Epidemiological data and gonococcal isolates were collected prospectively from patients at two UK clinics in London and Sheffield. Social and epidemiological data were combined with molecular typing of gonococcal isolates by a new methodology based on the polymorphisms of the opa gene. FINDINGS In London, interview data and opa-typing on samples from 215 cases showed a diverse population with few links. In Sheffield, interview data identified links between 51 (43%) of 120 cases, whereas opa-typing suggested a more connected population: 95 (79%) of cases had shared profiles. There was a highly significant correlation between the two distributions with epidemiological clusters appearing as a subset of the opa clusters. Two large opa clusters, of 18 and 43 cases, accounted for 50% of local cases of gonorrhoea. Discordance between epidemiological and opa-typing data was observed at highly connected points in the sexual network. INTERPRETATION Opa-typing is a more powerful tool for epidemiological investigation of gonorrhoea transmission than earlier methods. Opa-typing can link infections that would otherwise remain unlinked, and may aid interventions to control endemic disease.

The molecular diagnosis of lymphogranuloma venereum: evaluation of a real-time multiplex polymerase chain reaction test using rectal and urethral specimens.

Objectives: The objectives of this study were to evaluate the use of a real-time multiplex polymerase chain reaction (M-PCR) assay to differentiate between trachoma and lymphogranuloma venereum (LGV) biovars of Chlamydia trachomatis and to validate its performance with the conventional genotyping method. Study: Swab specimens from 115 patients with anorectal symptoms or syndromes associated with LGV were tested by a real-time M-PCR assay and the results compared with the PCR-based restriction fragment length polymorphism analysis of the major outer membrane protein gene (omp1). Results: A high agreement of 96.5% (111 of 115 specimens) was found between the real-time M-PCR testing and the standard genotyping method for the detection of C. trachomatis DNA (&kgr; value, 0.945, P <0.00001). Both methods identified 53 LGV, 32 non-LGV C. trachomatis, and 26 negative specimens. Conclusions: The real-time M-PCR assay simultaneously detects and differentiates LGV from non-LGV strains using swab specimens. This assay offers a relatively rapid and sensitive alternative for the diagnosis of LGV infection and is a useful tool for screening and for outbreak investigations.

Changing Epidemiologic Profile of Quinolone-Resistant Neisseria gonorrhoeae in London

The percentage of quinolone-resistant Neisseria gonorrhoeae isolated in London increased between 2000 and 2003, from 0.9% to 7.9% of total isolates. This increase was investigated by genotyping resistant isolates and comparing demographic and behavioral data. In 2000, resistant isolates predominantly had unique sequence types (STs) that were associated with imported infection, whereas, in 2002 and 2003, large ST clusters of indistinguishable isolates were associated with endemic acquisition. Resistant isolates that belonged to these large clusters were typically from patients who had similar epidemiological characteristics (such as ethnicity and sexual orientation) and behavioral characteristics (such as multiple sex partners and previous gonorrhea). In London, quinolone resistance is no longer associated with importation from areas of high prevalence and is spreading endemically in high-risk groups.

Rise in gonorrhoea in London, UK

Summary We report an alarming 34·9% overall increase in the number of gonococcal infections diagnosed during the same 3 month study period during 3 years. Between 1997 and 1998, an increase of 12·1% was detected, with a larger increase between 1998 and 1999, of 20·4%.

Concordance between Neisseria gonorrhoeae genotypes recovered from known sexual contacts.

ABSTRACT Neisseria gonorrhoeae multiantigen sequence typing (NG-MAST) is a highly discriminatory molecular typing procedure that provides precise and unambiguous strain characterization. Since molecular typing can complement contact tracing for reconstructing gonorrhea sexual networks, the concordance between the NG-MAST genotypes of pairs of N. gonorrhoeae isolates from recent sexual contacts was examined. Among 72 pairs of gonococci from recent sexual contacts, the genotypes of each pair were concordant in 65 cases (90.3%). In two further pairs, the isolates from sexual contacts differed by only a single nonsynonymous substitution in the porin gene, and in both of these pairs, the isolates were the same by opa typing. The other five nonconcordant pairs of isolates were clearly different strains. opa typing data were available for 51 of the pairs of isolates from sexual contacts, and concordant opa types were obtained in 38 cases (74.5%). NG-MAST should therefore be better than opa typing at identifying recent sexual contacts and has the important advantage over opa typing of being a more precise method of strain characterization.

Confirming the Chlamydia trachomatis status of referred rectal specimens

Objectives: To assess the reliability of different laboratory methods for the detection of Chlamydia trachomatis in rectal specimens Methods: 1782 rectal specimens confirmed as C trachomatis positive using a standard laboratory method, were forwarded to the Sexually Transmitted Bacteria Reference Laboratory (STBRL). All specimens were retested using a C trachomatis specific independent in-house real time polymerase chain reaction (PCR). If this test was negative, a second test (Artus Real-Art PCR Kit) was employed as a confirmation. A correlation between real time PCR results obtained at the reference centre (STBRL), and the method of C trachomatis detection used in the primary laboratory was undertaken. Results: The percentage of specimens that could be confirmed as positive, compared with primary method of detection was as follows: C trachomatis culture 87.5%, strand displacement assay (SDA: Becton Dickinson) 93.4%, Cobas Amplicor (Roche) 89.2%, Aptima Combo Two assay (Genprobe) 83.3%, and enzyme immunoassays (EIA) 35.4%. Conclusions: High rates of confirmation can be achieved using an independent real time PCR assay to examine rectal specimens which had initially tested C trachomatis positive using nucleic acid amplification tests and chlamydia tissue culture. This is not possible for specimens that had been screened using EIA tests, which reflects the low specificity of this test when used for rectal specimens. Laboratories currently using EIA based assays to test rectal specimens should review this approach.