Penicillin-resistant Neisseria Gonorrhoeae Research Articles

Draft Genome Sequences of Three Penicillin-Resistant Neisseria gonorrhoeae Strains Isolated in Cincinnati, Ohio, in 1994.

Here, we report the draft genome sequences of three penicillin-resistant Neisseria gonorrhoeae isolates. We include associated data on MICs and genetic relationships to other N. gonorrhoeae strains collected from across the United States. Resistance mutations known to contribute to reduced penicillin susceptibility are annotated in each genome.

Typing of Neisseria gonorrhoeae isolates by phenotypic and genotypic techniques in New Delhi, India.

BACKGROUND:The objective of this study is to investigate gonococcal isolates using phenotypic and genotypic methods.METHODOLOGY:Sixty gonococcal isolates obtained were examined. Strains were divided into 9 resistant phenotypes: Chromosomally mediated penicillin-resistant Neisseria gonorrhoeae (CMRNGP), penicillinase-producing NG (PPNG), chromosomally mediated tetracycline-resistant NG (CMRNGT), TRNG, PPNG and TRNG, CMRNGPT, quinolone resistant NG (QRNG), Azithro R, and decreased susceptibility (DS) to ceftriaxone. These isolates were also subjected to auxotyping and NG-multi-antigen sequence typing (MAST).RESULTS:Of 60 isolates, 32 (53.33%) PPNG and only one was CMRNGP; 16 (26.66%) were CMRNGT, while 18 (30%) were TRNG. Both PPNG and TRNG found in 13 (21.66%) and none were CMRNGPT. QRNG was seen in 93.33%, 5% Azithromycin R, and 6.66% were DS to ceftriaxone. Based on auxotyping, 24 (40%) nonrequiring, 16 (26.66%) were proline requiring, 13 (21.66%) arginine requiring while 7 (11.66%) belonged to others. The most common ST was 6058 (32.5%). The discriminatory indices of antibiogram, auxotyping and NG-MAST were 0.77, 0.72, and 0.95, respectively.CONCLUSIONS:NG-MAST is the method of choice for epidemiological studies.

Penicillin-binding protein 2 genes of chromosomally-mediated penicillin-resistant Neisseria gonorrhoeae from Greece: screening for codon Asp-345A.

Low affinity of penicillin for altered penicillin-binding protein 2 (PBP 2) of Neisseria gonorrhoeae that are chromosomally resistant to penicillin (CMRNG) has been attributed, at least partly to the insertion of an additional aspartic acid (Asp-35A) codon in the penA gene. Oligonucleotide probes were used to detect the presence or absence of this extra amino-acid residue in 34 non-penicillinase producing N. gonorrhoeae with decreased susceptibility to penicillin (MICs 0.25-2 mg/L) and 11 strains sensitive to penicillin (MIC < or = 0.06 mg/L) isolated in Greece and previously characterized by auxotype and serovar. The Asp-345A codon was found in all CMRNG strains as well as in strains with intermediate resistance to penicillin, but was absent from all strains exhibiting an MIC of penicillin of < or = 0.03 mg/L. No association of the Asp-345A insertion to any particular auxotype or serovar was found. These results expand the previously reported correlation between the presence of the Asp-345A codon and reduced sensitivity to penicillin, and shows that this particular mutation is common among epidemiologically distinct CMRNG strains.

Expression and purification of a soluble form of penicillin-binding protein 2 from both penicillin-susceptible and penicillin-resistant Neisseria gonorrhoeae

Resistance to penicillin in non-β-lactamase-producing strains of Neisseria gonorrhoeae (CMRNG strains) is mediated in part by the production of altered forms of penicillin-binding protein 2 (PBP 2) that have a decreased affinity for penicillin. The reduction in the affinity of PBP 2 is largely due to the insertion of an aspartic acid residue (Asp-345a) into the amino acid sequence of PBP 2. Truncated forms of N. gonorrhoeae PBP 2, which differed only by the insertion of Asp-345a, were constructed by placing the region of the penA genes encoding the periplasmic domain of PBP 2 (amino acids 42–581) into an ATG expression vector. When the recombinant PBP 2 molecules were over-expressed in Escherichia coli, insoluble PBP 2 inclusion bodies, which could be isolated by low-speed centrifugation of cell lysates, were formed. These insoluble aggregates were solubilized and the truncated PBP 2 polypeptides were partially purified by cation-exchange chromatography and gel filtration in the presence of denaturant prior to the refolding of the enzyme in vitro. After renaturation, gel filtration was used to separate monomeric soluble PBP 2 from improperly folded protein aggregates and other protein contaminants. A 4-liter culture of induced E. coli cells yielded 1.4 mg of soluble PBP 2 or PBP 2′ (PBP 2 containing the Asp-345a insertion), both of which were estimated to be 99% pure. The affinity of soluble PBP 2′ for [ 3H]penicillin G was decreased fourfold relative to that of soluble PBP 2, and their affinities were found to be identical to the affinities of the full-length PBP 2 enzymes that were previously determined in N. gonorrhoeae membranes. Furthermore, soluble PBP 2 displayed a rank order of affinity for several other β-lactam antibiotics that was consistent with the rank order of affinities previously reported for the native molecules. On the basis of these results, both of these soluble PBPs should be suitable for crystallization and X-ray crystallographic analysis.

Treatment of Uncomplicated Gonococcal Urethritis with Single-Dose Ceftizoxime

In areas such as Los Angeles, California, where penicillin-resistant Neisseria gonorrhoeae accounts for 1% or more of all gonorrhea strains isolated, third-generation cephalosporin agents have replaced penicillin as standard therapy. Ceftriaxone, the agent recommended by CDC guidelines, or ceftizoxime, an alternative agent have excellent activity against all strains of N. gonorrhoeae. One hundred seventy-five male patients with clinically and microbiologically confirmed gonococcal urethritis were treated with single-dose ceftizoxime 250 mg IM (50 patients) or 500 mg IM (125 patients). Cure was demonstrated clinically by repeat cultures of urethral discharge in all patients, including those with infections from penicillinase-producing strains of bacteria. None of the patients experienced complications or allergic reactions to ceftizoxime. Given the lower unit-of-use cost of ceftizoxime, its routine in place of ceftriaxone could result in substantial cost savings in those institutions that treat large numbers of patients with uncomplicated gonococcal urethritis.

Effect of penicillin and spectinomycin given for urethritis and cervicitis with Neisseria gonorrhoeae: high prevalence of penicillin-resistant isolates.

Efficacy of single-dose spectinomycin (TRO: 2 g intramuscularly) was compared with that of aqueous procaine penicillin G (APPG:4.8 x 10(6) units) plus 1 g of probenecid for treatment of gonococcal urethritis and cervicitis. Cure rates of the 210 patients who received TRO and 190 patients who received APPG were 97.6% and 91.1%, respectively. MICs of antibiotics were determined using the agar dilution method. Those isolates with MICs of APPG of less than 1.0 microgram/ml had low failure rates (2.9%), while strains with increased resistance to APPG (MICs greater than or equal to 1.0 microgram/ml) had higher failure rates (24%). Treatment failures seen with TRO were not correlated to isolates with the higher MICs. Clinical results suggest TRO could be given for treatment of genital gonorrhoea in Puerto Rico due to the high prevalence of both chromosomally-mediated penicillin-resistant Neisseria gonorrhoeae (20%) and penicillinase-producing Neisseria gonorrhoeae (7.5%) strains and the high rate of failure seen with the use of APPG.

Norfloxacin: clinical pharmacology and clinical use.

Norfloxacin, a nalidixic acid analog, is the first of the fluorinated quinolinecarboxylic acids to be marketed in the United States. It demonstrates potent antibacterial activity against aerobic, gram-negative bacteria including the Enterobacteriaceae, gentamicin-resistant Pseudomonas aeruginosa, and penicillin-resistant Neisseria gonorrhoeae. Norfloxacin exhibits good activity against methicillin-resistant and -sensitive Staphylococcus aureus, but less activity against most other aerobic, gram-positive organisms. Anaerobic bacteria are resistant to the drug. Resistance to norfloxacin is not plasmid mediated, but is secondary to bacterial mutation, and occurs less frequently than nalidixic acid resistance. Its pharmacokinetic properties after a 400-mg oral dose consist of a peak serum concentration of 1.3-1.58 micrograms/ml, an elimination half-life of 3-7 hours, and good penetration into kidney and prostatic tissues. Renal excretion is the major route of elimination. Norfloxacin is highly effective in the treatment of uncomplicated and complicated urinary tract infections, and gonococcal urethritis. Adverse effects are generally well tolerated and usually do not require discontinuation of therapy.

A community-based outbreak of infection with penicillin-resistant Neisseria gonorrhoeae not producing penicillinase (chromosomally mediated resistance): Faruki H, Kohmescher RN, McKinney WP, et al N Engl J Med 313:607–610 Sep 1985

A community-based outbreak of infection with penicillin-resistant Neisseria gonorrhoeae not producing penicillinase (chromosomally mediated resistance): Faruki H, Kohmescher RN, McKinney WP, et al N Engl J Med 313:607–610 Sep 1985

A community-based outbreak of infection with penicillin-resistant Neisseria gonorrhoeae not producing penicillinase (chromosomally mediated resistance) : Faruki H, Kohmescher RN, McKinney WP, et al: N Engl J Med 313:607–610, 1985

A community-based outbreak of infection with penicillin-resistant Neisseria gonorrhoeae not producing penicillinase (chromosomally mediated resistance) : Faruki H, Kohmescher RN, McKinney WP, et al: N Engl J Med 313:607–610, 1985

The nurse practitioner: history, current conflicts, and future survival.

“A Community-Based Outbreak of Infection With Penicillin-Resistant Neisseria Gonorrhoeae Not Producing Penicillinase (Chromosomally Mediated Resistance),” HAWAZIN FARUKI, et al. From February throu...

Lectin characterization of gonococci from an outbreak caused by penicillin-resistant Neisseria gonorrhoeae.

A total of 40 Neisseria gonorrhoeae isolates, representing 19 penicillin-resistant isolates (from 8 heterosexual patients and 11 homosexual patients) and 21 penicillin-susceptible isolates (from 15 heterosexual patients and 6 homosexual patients) and obtained from the same geographic area, were examined. Lectin agglutination patterns were based on the reactivity of the isolates with the following 14 lectins: concanavalin A, Lens culinaris, Trichosanthes kinlowii, Griffonia simplicifolia I, Arachis hypogeae (peanut agglutinin), Glycine max (soybean agglutinin), Dolichos bifloris, Griffonia simplicifolia II, Solanum tuberosum (potato starch agglutinin), Triticum vulgaris (wheat germ agglutinin), Limax flavus, Phaseolus vulgaris, Ulex europaeus I, and Lotus tetragonolobus. All isolates were serotyped with monoclonal antibodies specific for gonococcal outer membrane protein I and auxotyped, and the plasmid content was determined. Resistant patient isolates were selected for their decreased penicillin susceptibility, and control isolates were selected for their penicillin susceptibility. Even though the patient isolates demonstrated resistance to penicillin, no phenotypic differences in lectin-grouping patterns were demonstrated between the two study groups; i.e., two predominant lectin groups were observed. No resistance-associated plasmids were detected. All patient isolates were serogroup IB (serovars IB-1, IB-2, and IB-4), whereas 12 of 21 control isolates were serogroup IA (P less than 0.05). Isolates obtained from different anatomical sites in the same patient (cervical and rectal) agreed with regard to lectin patterns and serovars but not auxotypes.

A community-based outbreak of infection with penicillin-resistant Neisseria gonorrhoeae not producing penicillinase (chromosomally mediated resistance).

From February through November of 1983, 199 cases of penicillin-resistant Neisseria gonorrhoeae infection were identified in a localized epidemic in Durham, North Carolina. The isolates did not produce beta-lactamase but were unusually resistant to penicillin (minimum inhibitory concentration, 2.0 to 4.0 micrograms per milliliter), and 15 of 16 patients treated with 4.8 million units of penicillin G procaine plus 1.0 g of probenecid did not respond to therapy. Recognition of the outbreak was impeded by a lack of routine surveillance for resistance other than that mediated by beta-lactamase. All epidemic isolates had a single serotype, auxotype, and antibiotic-susceptibility profile. The outbreak was halted by changing the treatment for all patients and their contacts to spectinomycin, and by intensive epidemiologic case-finding efforts. The emergence of such resistant strains poses potential major public health problems and indicates a need for reassessment of current surveillance procedures.

Use of Non radioactive DNA Probes for the Detection of infectious Bacteria

Application of the biotinylated DNA probe systems in the detection of Chlamydia trachomatis and penicillin-resistant Neisseria gonorrhoeae is presented. The three formats employed are the dot-blot, in-situ, and Southern hybridization.

The Mental Health Walk-In Clinic: the University of Massachusetts experience.

Abstract A mental health walk-in clinic was introduced at the University of Massachusetts at Amherst in 1978 and was successful in maintaining accessible and early intervention services. Over the last few years, serious questions arose about the quality of service provided and the amount of stress on staff based on the volume of people needing to be seen during the walk-in clinic time period. A survey of 100 consecutive walk-in clients indentified a number of positive and negative factors including the fact that 43% of the people would have taken a regular 50-minute intake appointment if available within four days to two weeks. Several changes were implemented in the clinical intake system with very positive results. “Treatment of Penicillin-Resistant Neisseria Gonorrhoeae with Oral Norfloxacin,” STEVEN R. CRIDER, et al. Norfloxacin, an orally administered quinoline carboxylic acid that is structurally related to nalidixic acid, has been shown to be highly active in vitro against penicillinase-producing N...

ChemInform Abstract: 2‐ACETYLPYRIDINE THIOSEMICARBAZONES. XI: 2‐(α‐HYDROXYACETYL)PYRIDINE THIOSEMICARBAZONES AS ANTIMALARIAL AND ANTIBACTERIAL AGENTS

Abstract2‐Vinylpyridin (I) läßt sich mit Permanganat zum Glykol (II) hydroxylieren, das mit Br, das Hydroxyacetylpyridin (III) liefert;dieses ergibt mit Hydrazin (IV) das Hydrazon (V), an das sich die ′ Isothiocyanate (VI) zu den Thiosemicarbazonen (VII) addieren.

2-Acetylpyridine Thiosemicarbazones XI: 2-(α-Hydroxyacetyl)pyridine Thiosemicarbazones as Antimalarial and Antibacterial agents

A series of 2-(α-hydroxyacetyl)pyridine thiosemicarbazones was synthesized as potential antimalarial and antibacterial agents. Their synthesis was achieved by the condensation of N4-mono- or N4,N4-disubstituted thiosemicarbazides with 2-(α-hydroxyacetyl)pyridine. The latter was prepared by selective bromine oxidation of (2-pyridinyl)-1,2-ethanediol. The new compounds show potent inhibitory activity against penicillin-sensitive as well as penicillin-resistant Neisseria gonorrhoeae (MIC, 0.5–0.004 μg/mL), against Neisseria meningitidis (MIC, 0.5–0.032 μg/mL), and Staphylococcus aureus (MIC, 0.5–2 μg/mL). Good in vitro antimalarial effects against Plasmodium falciparum (Smith strain; ID50, 6.7–38 ng/mL) were observed in most of these new agents, but only 3 of 12 compounds exhibit moderate in vivo activity against Plasmodium berghei. These new agents appear to be less toxic to the host and more water soluble than the corresponding 2-acetylpyridine thiosemicarbazones.

In vitro antimicrobial activity of eight new beta-lactam antibiotics against penicillin-resistant Neisseria gonorrhoeae.

Increasing numbers of cases of penicillin-resistant gonorrhea necessitate the evaluation of new antibiotics for treatment of this disease. We tested the susceptibility of 92 penicillinase-producing (PP) Neisseria gonorrhoeae isolates and 88 penicillin-susceptible (PS) isolates to eight new beta-lactam antibiotics. The minimal inhibitory concentrations of these antibiotics were determined by the agar plate method. PP and PS N. gonorrhoeae isolates were susceptible to clinically achievable levels of all antibiotics tested. There were, however, marked differences among the drugs with regard to the concentration required to inhibit growth. The PP N. gonorrhoeae isolates were extremely susceptible to ceftriaxone, ceftizoxime, and cefotaxime, highly susceptible to moxalactam and cefoperazone, and less susceptible to cefoxitin, ceforanide, and cefonicid (geometric mean minimal inhibitory concentrations were 0.002, 0.003, 0.007, 0.03, 0.07, 0.6, 2.4, and 3.1 micrograms/ml, respectively). Although this in vitro study showed PP N. gonorrhoeae isolates to be comparatively more susceptible to ceftriaxone, ceftizoxime, and cefotaxime than to the other antibiotics, these results may not correlate with clinical efficacy.

Resistance to penicillin and identification of penicillinase-producing Neisseria gonorrhoeae among clinical isolates in Thailand.

Penicillin-resistant Neisseria gonorrhoeae from 405 patients were studied by determination of penicillin minimal inhibitory concentrations an penicillinase production. Eighteen percent were identified as penicillin-producing N. gonorrhoeae and the mean minimal inhibitory concentration, for all except penicillin-producing strains, was 0.805 micrograms/ml.

Penicillin-resistant gonococcal polyarthritis

PENICILLIN has been the preferred treatment of gonococcal infections for years. However, in 1976, reports from Great Britain and the United States described patients with uncomplicated genitourinary infections due to penicillin-resistant Neisseria gonorrhoeae , which produced β-lactamase (penicillinase). 1,2 We describe a patient with infection of the pharynx and destructive polyarthritis due to a β-lactamase—producing strain of N gonorrhoeae . Report of a Case A 25-year-old man was admitted to a US Navy Hospital in the Philippine Islands on Aug 15, 1976, with a two-day history of fever, migratory arthritis, and transient red papules on his limbs. He had an effusion of the left knee, with a WBC count of 14,300/cu mm, with 85% polymorphonuclear leukocytes. Gram's stain and cultures of the synovial fluid were negative, and roentgenograms of his joints were normal. He was treated with oral penicillin G potassium, salicylates, and prednisone for possible acute rheumatic fever. Temperatures of 38