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  • 1.
    Olsen, Birgitta
    et al.
    Örebro University, Department of Clinical Medicine.
    Hadad, Ronza
    Fredlund, Hans
    Örebro University, Department of Clinical Medicine.
    Unemo, Magnus
    Örebro University, Department of Clinical Medicine.
    A view of the Neisseria gonorrhoeae population in Sweden in 2005: phenotypic and genetic characterisation2006Conference paper (Other academic)
    Abstract [en]

    Aims

    To phenotypically and genotypically characterise clinical N. gonorrhoeae isolates transmitted in Sweden during 2005 and to compare with characteristics of N. gonorrhoeae populations in other countries.

    Introduction

    In Sweden, the gonorrhoea incidence decreased from the beginning of the 1970s to 1996. From 1997 the incidence has almost annually increased, mainly due to a rise in domestic cases of young heterosexuals of both sexes and homosexual men. Furthermore, during recent decades resistance to most of the traditional antibiotics used in the treatment of gonorrhoea has rapidly increased worldwide. Availability of effective diagnostics, treatment and surveillance of epidemiological characteristics (antibiotic susceptibility, serovars and genotypes) are main tools for control of the transmission of infection.

    Materials, Methods & Results

    N. gonorrhoeae isolates (n=175) cultured in Sweden in 2005 and received at the Swedish Reference Laboratory for Pathogenic Neisseria were included. Phenotypic characterisation was performed by antibiotic susceptibility testing and serovar determination using both Genetic systems (GS) and Pharmacia (Ph) panels of monoclonal antibodies (MAbs). Genetic characterisation was performed using N. gonorrhoeae multiantigen sequence typing (NG-MAST) that analyses more variable segments of the porB gene (490 bp) and of the tbpB gene (390 bp).

    All isolates were susceptible to cefixime, ceftriaxone, and spectinomycin. The levels of intermediate susceptibility and resistance to azithromycin, ciprofloxacin, and ampicillin were 2.3%, 49.7% and 75.3%, respectively (Table 1). Fifty-seven (33%) of the isolates were -lactamase producing.

    In total, 33 of the isolates were determined as serogroup WI (PorB1a). These were assigned nine different GS-serovars and seven different Ph-serovars. Two of the PorB1a isolates were not serotypeable using Ph MAbs. The remaining 142 isolates were determined as serogroup WII/III (PorB1b). These isolates were assigned 18 different GS-serovars and 38 different Ph-serovars.

    The isolates displayed 66 and 56 divergent NG-MAST porB and tbpB alleles, respectively. These resulted in assignment of 95 different sequence types (STs), of which 34 have not been previously described. ST40 (n=15), ST225 (n=12), ST1813 (n=9), ST5 (n=8), ST753 (n=6), ST323 (n=5), and ST211 (n=4), were the most prevalent STs. Four STs were represented by three isolates, 20 STs by two isolates and 64 STs by single isolates (Figure 1).

    Conclusions

    A highly diverse N. gonorrhoeae population was transmitted in Sweden during 2005, which can reflect importation of strains from abroad, and/or in some geographic areas suboptimal diagnostics and incomplete epidemiological surveillance. However, many clusters of isolates, which can reflect the existence of several transmission chains, were also identified.

    Serovar determination is still fairly effective, rapid, inexpensive, easily performed and remains a valuable primary epidemiological marker of N. gonorrhoeae, which can supplement the superior genetic characterisation.

  • 2.
    Olsen, Birgitta
    et al.
    Örebro University, School of Health and Medical Sciences.
    Hadad, Ronza
    Fredlund, Hans
    Örebro University, School of Health and Medical Sciences.
    Unemo, Magnus
    Örebro University, School of Health and Medical Sciences.
    The Neisseria gonorrhoeae population in Sweden during 2005: phenotypes, genotypes and antibiotic resistance2008In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 116, no 3, p. 181-189Article in journal (Refereed)
    Abstract [en]

    In Sweden, the gonorrhoea incidence has significantly increased since an all-time low in 1996. We aimed to phenotypically and genotypically characterise N. gonorrhoeae isolates (nΩ180) transmitted in Sweden during 2005. All isolates were susceptible to cefixime, ceftriaxone, and spectinomycin. However, 2%, 50% and 75% displayed intermediate susceptibility or resistance to azithromycin, ciprofloxacin and ampicillin, respectively. The isolates were assigned to 28 different serovars using Genetic Systems monoclonal antibodies (Mabs) (discriminatory index, 91.0%) and 46 different serovars using Pharmacia Mabs (index, 94.4%). Furthermore, they displayed 95 porB sequences (index, 97.8%) and 95 N. gonorrhoeae multiantigen sequence typing (NG-MAST) sequence types (STs) (index, 98.0%). 51 (54%) of these STs have not been previously described. 14 ST clusters, comprising between 3 and 15 isolates, were identified that indicate the existence of several transmission chains. The high number of unique STs (nΩ63) may be associated with import of strains from abroad, local emergence of new STs, incomplete epidemiological surveillance, and/or suboptimal diagnostics, including contact tracing. Overall, the Swedish N. gonorrhoeae population was remarkably diversified. Comprehensive knowledge regarding transmission, phenotypes (including antibiotic resistance), but also in many cases highly discriminative and precise genotypic characteristics of the N. gonorrhoeae strains circulating in our societies, is crucial.

  • 3.
    Olsen, Birgitta
    et al.
    Örebro University, Department of Clinical Medicine.
    Lan, P. T.
    Stålsby Lundborg, Cecilia
    Tran, Hau Khang
    Unemo, Magnus
    Örebro University, Department of Clinical Medicine.
    First ever population-based assessment of Mycoplasma genitalium in Vietnam: low prevalence among married women of reproductive age in rural areas2008Conference paper (Other academic)
    Abstract [en]

    Objective: To analyse the prevalence of Mycoplasma genitalium infection in a population-based study among sexually active married women from a demographic surveillance site in a rural geographical area of Vietnam.

    Materials and Methods: Women, 18-49 years of age, were randomly selected to participate. DNA was isolated from endocervical swabs sampled from 990 participating women. The M. genitalium MgPa adhesion gene was detected using a real-time PCR with TaqMan probe

    Results: Eight (0.8% [95% confidence interval, 0.25-1.35%]) of the included women were infected with M. genitalium. Two of these positive women reported clinical symptoms. One additional M. genitalium positive but symptom-free woman, however, showed clinical signs of vaginitis. None of the M. genitalium positive women was concomitantly infected with Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum or HIV. Furthermore, there was no obvious association between M. genitalium infection and vaginal douching, use of intra-uterine device (IUD), or occurrence of bacterial vaginosis, candidiasis, or Trichomonas vaginalis.

    Conclusions: The prevalence of M. genitalium among sexually active married women in Vietnam was relatively low. However, more large, well-designed and appropriately performed studies in other population groups including unmarried women and men, and in other geographical areas, rural as well as urban, are crucial in order to extract any evidence-based conclusions regarding the overall prevalence of STIs, including M. genitalium infections, in the Vietnamese society. The present study compiled with such future studies may form the basis for a national sexual health strategy for prevention, diagnosis, and surveillance of STIs, including M. genitalium infections, in Vietnam.

  • 4.
    Olsen, Birgitta
    et al.
    Örebro University, School of Health and Medical Sciences.
    Lan, Pham T.
    Stålsby Lundborg, Cecilia
    Khang, T. H.
    Unemo, Magnus
    Örebro University, School of Health and Medical Sciences.
    Population-based assessment of Mycoplasma genitalium in Vietnam: low prevalence among married women of reproductive age in a rural area2009In: Journal of the European Academy of Dermatology and Venereology, ISSN 0926-9959, E-ISSN 1468-3083, Vol. 23, no 5, p. 533-537Article in journal (Refereed)
    Abstract [en]

    Objective To analyse the prevalence of Mycoplasma genitalium infection in a population-based study among married women from a demographic surveillance site in a rural geographical area of Vietnam.

    Materials and Methods Women, 18-49 years of age, were randomly selected to participate. DNA was isolated from endocervical swabs sampled from 990 participating women. The M. genitalium MgPa adhesion gene was detected using a real-time PCR with TaqMan probe.

    Results Eight (0.8% [95% confidence interval, 0.25-1.35%]) of the included women were infected with M. genitalium .Two of these positive women reported clinical symptoms. One additional M. genitalium positive but symptom-free woman, however, showed clinical signs of vaginitis. None of the M. genitalium positive women was concomitantly infected with Chlamydia trachomatis , Neisseria gonorrhoeae , syphilis or HIV. Furthermore, there was no obvious association between M. genitalium infectionand vaginal douching, use of intra-uterine device (IUD), or occurrence of bacterial vaginosis, candidiasis, or Trichomonas vaginalis .

    Conclusions The prevalence of M. genitalium among married women in Vietnam was relatively low. However, more large, well-designed and appropriately performed studies in other population groups including unmarried women and men, and in other geographical areas, rural as well as urban, are crucial in order to extract any evidence-based conclusions regarding the overall prevalence of STIs, including M. genitalium infections, in the Vietnamese society. The present study compiled with such future studies may form the basis for a national sexual health strategy for prevention, diagnosis, and surveillance of STIs, including M. genitalium infections, in Vietnam.

  • 5.
    Olsen, Birgitta
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Månsson, Fredrik
    Infectious Diseases Research Unit, Department of Clinical Sciences, Lund, University, Malmö, Sweden.
    Camara, Cidia
    National Public Health Laboratory (LNSP), Bissau, Guinea-Bissau.
    Monteiro, Mario
    National Public Health Laboratory (LNSP), Bissau, Guinea-Bissau.
    Biai, Ansu
    National Public Health Laboratory (LNSP), Bissau, Guinea-Bissau.
    Alves, Alfredo
    Department of Obstetrics and Gynaecology, Simaõ Mendes National Hospital, Bissau, Guinea-Bissau.
    Andersson, Sören
    WHO Collaborating Centre for Gonorrhoea and other STIs, National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden .
    Norrgren, Hans
    Division of Infection Medicine, Department of Clinical Sciences, Lund University, Lund, Sweden.
    Unemo, Magnus
    WHO Collaborating Centre for Gonorrhoea and other STIs, National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
    Phenotypic and genetic characterisation of bacterial sexually transmitted infections in Bissau, Guinea-Bissau, West Africa: a prospective cohort study2012In: BMJ Open, ISSN 2044-6055, E-ISSN 2044-6055, Vol. 20, no 2, article id e000636Article in journal (Refereed)
    Abstract [en]

    Background: Knowledge regarding characteristics and transmission of Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium and antibiotic resistance in N gonorrhoeae in Guinea-Bissau, West Africa, is entirely lacking.

    Objectives: To characterise N gonorrhoeae, C trachomatis and M genitalium samples from Guinea-Bissau and to define bacterial populations, possible transmission chains and for N gonorrhoeae spread of antibiotic-resistant isolates.

    Design: Prospective cohort study.

    Setting: Two sexual health and family planning clinics, Bissau, Guinea-Bissau.

    Participants: Positive samples from 711 women and 27 men.

    Material and methods: Positive samples for N gonorrhoeae (n=31), C trachomatis (n=60) and M genitalium (n=30) were examined. The gonococcal isolates were characterised with antibiograms, serovar determination and N gonorrhoeae multiantigen sequence typing (NG-MAST). The C trachomatis ompA gene and the M genitalium mgpB gene were sequenced, and phylogenetic analyses were performed.

    Results: For N gonorrhoeae, the levels of resistance (intermediate susceptibility) to ciprofloxacin, erythromycin, rifampicin, ampicillin, tetracycline, penicillin G and cefuroxime were 10% (0%), 6% (10%), 13% (10%), 68% (0%), 74% (0%), 68% (16%) and 0% (84%), respectively. All isolates were susceptible to cefixime, ceftriaxone, spectinomycin and azithromycin, and the minimum inhibitory concentrations of kanamycin (range: 8-32 mg/l) and gentamicin (range: 0.75-6 mg/l) were low (no resistance breakpoints exist for these antimicrobials). 19 NG-MAST sequence types (STs) (84% novel STs) were identified. Phylogenetic analysis of the C trachomatis ompA gene revealed genovar G as most prevalent (37%), followed by genovar D (19%). 23 mgpB STs were found among the M genitalium isolates, and 67% of isolates had unique STs.

    Conclusions: The diversity among the sexually transmitted infection (STI) pathogens may be associated with suboptimal diagnostics, contact tracing, case reporting and epidemiological surveillance. In Guinea-Bissau, additional STI studies are vital to estimate the STI burden and form the basis for a national sexual health strategy for prevention, diagnosis and surveillance of STIs.

  • 6.
    Olsen, Birgitta
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Thi Lan, Pham
    Department of Dermato-Venerology, Hanoi Medical University, Hanoi, Vietnam; National Hospital of Dermatology and Venerology, Hanoi, Vietnam.
    Golparian, Daniel
    WHO Collaborating Centre for Gonorrhoea and Other STIs, National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital, Örebro, Sweden.
    Johansson, Emma
    WHO Collaborating Centre for Gonorrhoea and Other STIs, National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital, Örebro, Sweden.
    Khang, Tran Hau
    Department of Dermato-Venerology, Hanoi Medical University, Hanoi, Vietnam; National Hospital of Dermatology and Venerology, Hanoi, Vietnam.
    Unemo, Magnus
    Örebro University Hospital. Örebro University, School of Medical Sciences. WHO Collaborating Centre for Gonorrhoea and Other STIs, National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Clinical Microbiology.
    Antimicrobial susceptibility and genetic characteristics of Neisseria gonorrhoeae isolates from Vietnam, 20112013In: BMC Infectious Diseases, ISSN 1471-2334, E-ISSN 1471-2334, Vol. 13, no 1, article id 40Article in journal (Refereed)
    Abstract [en]

    BACKGROUND: Antimicrobial resistance (AMR) in Neisseria gonorrhoeae is a major public health concern worldwide. In Vietnam, knowledge regarding N. gonorrhoeae prevalence and AMR is limited, and data concerning genetic characteristics of N. gonorrhoeae is totally lacking. Herein, we investigated the phenotypic AMR (previous, current and possible future treatment options), genetic resistance determinants for extended-spectrum cephalosporins (ESCs), and genotypic distribution of N. gonorrhoeae isolated in 2011 in Hanoi, Vietnam.

    METHODS: N. gonorrhoeae isolates from Hanoi, Vietnam isolated in 2011 (n = 108) were examined using antibiograms (Etest for 10 antimicrobials), Neisseria gonorrhoeae multi-antigen sequence typing (NG-MAST), and sequencing of ESC resistance determinants (penA, mtrR and penB).

    RESULTS: The levels of in vitro resistance were as follows: ciprofloxacin 98%, tetracycline 82%, penicillin G 48%, azithromycin 11%, ceftriaxone 5%, cefixime 1%, and spectinomycin 0%. The MICs of gentamicin (0.023-6 mg/L), ertapenem (0.002-0.125 mg/L) and solithromycin (<0.016-0.25 mg/L) were relatively low. No penA mosaic alleles were found, however, 78% of the isolates contained an alteration of amino acid A501 (A501V (44%) and A501T (34%)) in the encoded penicillin-binding protein 2. A single nucleotide (A) deletion in the inverted repeat of the promoter region of the mtrR gene and amino acid alterations in MtrR was observed in 91% and 94% of the isolates, respectively. penB resistance determinants were detected in 87% of the isolates. Seventy-five different NG-MAST STs were identified, of which 59 STs have not been previously described.

    CONCLUSIONS: In Vietnam, the highly diversified gonococcal population displayed high in vitro resistance to antimicrobials previously recommended for gonorrhoea treatment (with exception of spectinomycin), but resistance also to the currently recommended ESCs were found. Nevertheless, the MICs of three potential future treatment options were low. It is essential to strengthen the diagnostics, case reporting, and epidemiologic surveillance of gonorrhoea in Vietnam. Furthermore, the surveillance of gonococcal AMR and gonorrhoea treatment failures is imperative to reinforce. Research regarding novel antimicrobial treatment strategies (e.g., combination therapy) and new antimicrobials is crucial for future treatment of gonorrhoea.

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