Antimicrobial susceptibility and genetic characteristics of Neisseria gonorrhoeae isolates from India, Pakistan and Bhutan in 2007–2011

BMC Infectious Diseases (Impact Factor: 2.61). 01/2013; 13(1):35. DOI: 10.1186/1471-2334-13-35
Source: PubMed


Knowledge on antimicrobial drug resistance and genetic characteristics of Neisseria gonorrhoeae isolates circulating in India, Pakistan, and Bhutan is sorely lacking. In this paper, we describe the prevalence of antimicrobial resistance and molecular characteristics of N. gonorrhoeae isolates from India, Pakistan, and Bhutan in 2007–2011.

Antimicrobial susceptibility and β-lactamase production were tested for 65 N. gonorrhoeae isolates from India (n=40), Pakistan (n=18) and Bhutan (n=7) using Etest methodology (eight antimicrobials) and nitrocefin solution, respectively. Resistance determinants, i.e. penA, mtrR, porB1b, gyrA, and parC, were sequenced. N. gonorrhoeae multiantigen sequence typing (NG-MAST) was performed for molecular epidemiology.

The highest resistance level was observed for ciprofloxacin (94%), followed by penicillin G (68%), erythromycin (62%), tetracycline (55%), and azithromycin (7.7%). All the isolates were susceptible to ceftriaxone, cefixime, and spectinomycin. Thirty-four (52%) of the isolates were producing β-lactamase. No penA mosaic alleles or A501-altered alleles of penicillin-binding protein 2 were identified. Forty-nine NG-MAST STs were identified, of which 42 STs have not been previously described worldwide.

Based on this study, ceftriaxone, cefixime, and spectinomycin can be used as an empirical first-line therapy for gonorrhoea in India, Pakistan, and Bhutan, whereas ciprofloxacin, penicillin G, tetracycline, erythromycin, and azithromycin should not be. It is imperative to strengthen the laboratory infrastructure in this region, as well as to expand the phenotypic and genetic surveillance of antimicrobial resistance, emergence of new resistance, particularly, to extended-spectrum cephalosporins, and molecular epidemiology.

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    • "CEF is chemically known as (6R, 7R)-7-{[2-(2-amino-1,3-thiazol- 4-yl)2(carboxymethoxyimino) acetyl]amino}-3-ethenyl-8- oxo-5-thia-1-azabicyclo[4.2.0] oct-2-ene-2-carboxylic acid (Figure 1B) (Ali Ahmed et al., 2013). CEF is the extendedspectrum cephalosporins, are currently the recommended first-line antimicrobials in most countries worldwide, can be used for the treatment of gonorrhea as an empirical first-line therapy (Sethi et al., 2013 and Olsen et al., 2013). Synthesis of cell wall inhibit by the bactericidal action of cephalosporin, which inhibits the final transpeptidation step of the peptidoglycan synthesis in the bacterial cell wall, thus inhibiting biosynthesis of cell wall assembly by arresting, resulting in bacterial cell death. "
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    ABSTRACT: The objective of the current study was to develop and validate a simple, accurate, precise and selective stability-indicating gradient reverse phase high performance liquid chromatographic method for simultaneous estimation of Ofloxacin and Cefixime in pharmaceutical formulation in presence of degradation products. The chromatographic separation of Ofloxacin and Cefixime was achieved on Shimadzu LC-20AT series HPLC having C18-ODS bonded column (250 x 4.6 mm, 40 °C, 10 µL) using UV/Visible detector at 276 nm. The optimized mobile phase was consisted of a methanol: phosphate buffer (50:50) at a flow rate of 1.0 ml/m. The retention times were 4.799 and 1.602 m for Ofloxacin and Cefixime respectively. The proposed method provided linear responses within the concentration ranges 5-25 µg/ml for Ofloxacin and Cefixime both. The limit of detection (LOD) and limit of quantification (LOQ) values were found to be 0.0259, 0.078 µg/ml and 0.0206, 0.062 µg/ml for Ofloxacin and Cefixime F respectively. The developed method was validated as per ICH guidelines with respect to specificity, linearity, accuracy, precision, robustness and ruggedness. The studies data revealed that developed method was convenient, fairly reliable, sensitive, less expensive and reproducible.
    Preview · Article · Feb 2015 · Science, Technology and Arts Research Journal
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    • "Gonorrhoea, which is caused by N. gonorrhoeae, is a common sexually transmitted infection (STI) that is associated with high morbidity and significant socioeconomic consequences. N. gonorrhoeae has repeatedly demonstrated the ability to develop resistance to all antimicrobial drugs introduced as first-line therapy [4]. Surveillance of both of these infectious diseases is critical for an accurate estimation of disease incidence and for appropriate prevention and treatment [5]. "
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    ABSTRACT: Background Although sophisticated methodologies are available, the use of endpoint polymerase chain reaction (PCR) to detect 16S rDNA genes remains a good approach for estimating the incidence and prevalence of specific infections and for monitoring infections. Considering the importance of the early diagnosis of sexually transmitted infections (STIs), the development of a sensitive and affordable method for identifying pathogens in clinical samples is needed. Highly specific and efficient primers for a multiplex polymerase chain reaction (m-PCR) system were designed in silico to detect the 16S rDNA genes of four bacteria that cause genital infections, and the PCR method was developed. Methods The Genosensor Probe Designer (GPD) (version 1.0a) software was initially used to design highly specific and efficient primers for in-house m-PCR. Single-locus PCR reactions were performed and standardised, and then primers for each locus in turn were added individually in subsequent amplifications until m-PCR was achieved. Amplicons of the expected size were obtained from each of the four bacterial gene fragments. Finally, the analytical specificity and limits of detection were tested. Results Because they did not amplify any product from non-STI tested species, the primers were specific. The detection limits for the Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum primer sets were 5.12 × 105, 3.9 × 103, 61.19 × 106 and 6.37 × 105 copies of a DNA template, respectively. Conclusions The methodology designed and standardised here could be applied satisfactorily for the simultaneous or individual detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum. This method is at least as efficient as other previously described methods; however, this method is more affordable for low-income countries.
    Full-text · Article · Jul 2014 · BMC Research Notes
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    • "Exceedingly high prevalence of resistance was observed for previous first-line antimicrobials such as ciprofloxacin (98%), tetracycline (82%) and penicillin G (48%), but also relatively high for azithromycin (11%) (Table 1). This is in accordance with previous studies from other countries in WHO WPR such as Japan, The Philippines, China, Hong Kong, Korea and Taiwan [4,46-51], South Asia, e.g., India, Pakistan, Thailand, Sri Lanka and Bhutan [47,52,53], and many other regions globally [3-6,18,19,54-58]. None of these antimicrobials can be recommended for first-line empiric therapy of gonorrhoea in Vietnam as well as in most parts of the world. "
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    ABSTRACT: 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.
    Full-text · Article · Jan 2013 · BMC Infectious Diseases
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