Identification of the Gonococcus from Cultures and the Effect of Certain Animal Sera on the Fermentations of the Gonococcus.
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Journal Article↗
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Gonococcus antigen possessing excessive anticomplementary activity; a method for preparing a specific antigen from a strongly anticomplementary dried gonococcus antigen.
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Antibodies↗
Identification of Neisseria gonorrhoeae in cultures from tonsillo-pharyngeal specimens by means of a slide co-agglutination test (Phadebact Gonococcus Test).
A slide co-agglutination test (Phadebact Gonococcus Test) for the serological identification of Neisseria gonorrhoeae was assessed on gonococcal-like, oxidase positive colonies from 120 cultures, originating from about 6,500 consecutive tonsillo-pharyngeal specimens received at the Neisseria Department, Statens Seruminstitut. The test was performed after subculture on a serum-free medium, since this procedure was found to reduce the number of strains showing inconclusive reactions (pseudo co-agglutination). If this pseudo co-agglutination does occur, however, the test can be repeated with the addition of trypsin to the test system. This causes the previously inconclusive reactions to be reverted to clearly positive reactions in the case of gonococci, and to clearly negative reactions in more than half of the previously inconclusive reactions with other bacterial strains. The results obtained by the Phadebact Gonococcus Test were compared with those obtained by bacteriological identification procedures. Fifty-six of the 120 cultures examined contained gonococci, and all strains were identified by the slide co-agglutination test (five strains with the addition of trypsin). The remaining 64 cultures were negative or exhibited consistently pseudo co-agglutination (eight strains). The specificity and sensitivity of the reagent was further confirmed by the examination of 53 strains of Neisseria gonorrhoeae and 50 strains representing Neisseria species commonly occurring in tonsillo-pharyngeal specimens. The Phadebact Gonococcus Test was considered to be a reliable alternative to routine bacteriological identification of Neisseria gonorrhoeae.
Agglutination Tests↗
The biology of the gonococcus.
Gonorrhea has been known since antiquity. Today, this disease is the most commonly reported infectious disease in the U.S. The natural environment of the etiological agent, Neisseria gonorrhoeae, is man. In this host, the organism usually parasitizes mucosal surfaces populated by columnar epithelial cells. Under certain conditions, the gonococcus may disseminate or spread to adjacent organs. The gonococcus is well adapted to its environment and is a successful parasite. Until recently, gonococci were uniformly sensitive to penicilin. However, a plasmid encoding beta-lactamase has been identified in some isolates. Most strains exhibit specific requirements for various amino acids, vitamins, purines, and pyrimidines. Only glucose, pyruvate, and lactate are utilized as sources of energy. Glucose is dissimilated by a combination of the Entner-Doudoroff and pentose phosphate pathways. A tricarboxylic acid cycle is also present and active under certain conditions. Structurally, the cell envelope of the gonococcus resembles that of a typical Gram-negative bacterium. Gonococci are highly autolytic, especially in older cultures or after depletion of the energy source. Autolysis is not due solely to peptidoglycan hydrolysis, but appears to involve a destabilization of the outer membrane as well. Cell surface components such as pili, lipopolysaccharide, outer membrane proteins, and a capsule are associated with the virulence and pathogenicity of this organism.
Amino Acids↗
Degeneration of the Gonococcus (in various media).
The ultrastructure of the gonococcus cultivated in different synthetic media and inoculated into the peritoneal cavity of the mouse makes it possible to distinguish the various degenerative morphological states of the microbe. A comparative study of phagocytosis and transformation of the gonococcus by leukocytes together with the changes observed in artificial media discloses a certain resemblance between the different stages of the progression towards L forms. Reversion to normal forms is only possible if gonococcal degeneration has not reached an extreme point and if the new medium compensates for the unfavourable conditions which produced L transformation.
Animals↗
Beta-lactamase-producing, penicillin-resistant gonococcus.
A gonococcus was isolated from the vagina of a woman with pelvic inflammatory disease. The organism was typical in all respects except that, unlike any previously described gonococcus, it produced a beta-lactamase that conferred resistance to penicillin, ampicillin, and cephaloridine. Among other antibiotics tested streptomycin was the only one to which the organism was resistant.
Ampicillin↗
Evaluation of the Phadebact Gonococcus Test for confirmation of Neisseria gonorrhoeae.
Strains of Neisseria gonorrhoeae grown on Thayer-Martin medium and media with and without serum were examined by the Phadebact Gonococcus Test. By using the direct method, in which colonies from growth media were mixed directly with the reagents on a microscope slide, Thayer-Martin medium, contrary to the manufacturer's claim, was not found to be the medium giving best results, and presence or absence of serum in the media did not affect the results. A large number of inconclusive results were obtained. A modification of the alternative procedure, which uses a heated suspension of the gonococcal cells as the antigen, is described. Of the 432 strains of N. gonorrhoeae examined by this method, 427 were accurately identified. Five strains, however, gave false-negative results. None of the 80 strains of N. meningitidis gave positive results. Nine of nineteen strains of N. lactamica, however, gave clear-cut positive results. Use of the o-nitrophenyl-beta-D-galactopyranoside test in conjunction with the Phadebact Gonococcus Test, particularly on isolates from the pharynx, is recommended.
Agglutination↗
Sensitivity of gonococcus strains isolated in Rotterdam to epicillin, ampicillin, penicillin and tetracycline.
Quantitative determinations of the sensitivity to epicillin, ampicillin, penicillin and tetracycline were carried out for 67 gonococcus strains isolated from 67 males with urogenital gonorrhoea, and 38 strains isolated from 38 females with urogenital/rectal gonorrhoea. The results of the sensitivity determinations for ampicillin, penicillin and tetracycline were compared with the findings of a previous investigation. It was found that the gonococcus strains isolated from males had become slightly less sensitive to penicillin between 1971-1972 and 1976. The penicillin and tetracycline had not changed during this period. It was also striking that the in vitro activity of epicillin for strains isolated both from males and females was significantly greater than that of ampicillin.
Ampicillin↗
The capsule of the gonococcus.
16 strains of Neisseria gonorrhoeae were subjected to several established techniques for capsule demonstration by light microscopy. In all strains examined, encapsulation of the gonococcus was demonstrated. Although the capsules were somewhat more easily seen in strains recently isolated from clinical material, organisms that had been passaged for several years also were encapsulated as were all the colony types within these strains. The gonococcal capsule is easily disrupted by mechanical forces.
Microscopy↗
Studies on gonococcus infection. VII. In vitro killing of gonococci by human leukocytes.
The comparative killing of pilated and nonpilated forms of Neisseria gonorrhoeae by human peripheral blood leukocytes was studied in vitro. Some nonpilated gonococci (T2) were killed to a lesser extent than were pilated, T2 organisms, which were killed less readily than another nonpilated (T4*) form of gonococcus. Thus, the relative order of killing of gonococci by human peripheral blood leukocytes appears to be: T4 less than T2 less than T4*. These data suggest that pilation, though correlated with virulence of gonococci, has little influence on the survival or killing of these organisms by human leukocytes.
Bacteriological Techniques↗
Studies on gonococcus infection. XIII. Occurrence of color/opacity colonial variants in clinical cultures.
Colonial variants of Neisseria gonorrhoeae differing in color and opacity characteristics are distributed differently between male urethral and female cervical cultures. Colonial characteristics of cultures isolated from female cervices differ with time of isolation within the menstrual cycle and use of oral contraceptives. These differences may reflect selective forces in the ecology of the cervix, particularly proteolytic enzymes in cervical mucus and menstrual blood. Cycle changes in the characteristics of gonococci isolated from females may have implications for pathogenesis of gonococcal infections and immune response to the gonococcus.
Cervix Uteri↗
Cloning and characterization of the catalase gene of Neisseria gonorrhoeae: use of the gonococcus as a host organism for recombinant DNA.
The structural gene for the catalase of Neisseria gonorrhoeae was cloned into a Kat- strain of that organism by using a recombinant vector derived from one of the beta-lactamase-specifying plasmids found in that organism. The kat gene was then successfully subcloned into both pUC8 and pGB2, transformed into Escherichia coli, and shown to complement the E. coli katE mutants UM2 and UMRl. The gene was subsequently mutagenized and returned to the gonococcus to generate a Kat- strain that was phenotypically identical to the strain originally used to clone the gene. The sequence of the gene and the derived amino acid sequence showed that the gonococcal kat gene closely resembles the hktE gene of Haemophilus influenzae. The sequence of the promoter region of the gonococcal kat gene is unusual and may explain the extremely high, loosely regulated expression of the gene.
Amino Acid Sequence↗
Evaluation of the phadebact gonococcus test, a coagglutination procedure for confirmation of Neisseria gonorrhoeae.
Rapid and accurate immunological confirmation of presumptively positive gonococci could be facilitated with the Phadebact Gonococcus Test, a slide coagglutination procedure. The test was compared with carbohydrate utilization and fluorescent-antibody tests on 235 clinical isolates. With the coagglutination procedure, 97.1% of the isolates were identified as compared with 93.1% by carbohydrate utilization and 98.7% by fluorescent antibody. The Phadebact test was highly specific, showing no cross-reactions with 55 other Neisseria species or with 50 miscellaneous organisms occasionally found growing on selective culture media. Because of its high sensitivity and specificity, ease of performance, and ability to provide results in 2 to 3 min, this procedure provides a suitable alternative to the carbohydrate utilization and fluorescent-antibody tests for confirmation of N. gonorrhoeae.
Agglutination Tests↗
Comparison of the Phadebact Gonococcus Test with the rapid fermentation method.
The Phadebact Gonococcus Test (Pharmacia Diagnostics, Piscataway, N.J.), a coagglutination technique, was compared with the rapid fermentation method of Kellogg and Turner (D. S. Kellogg, Jr., and E. M. Turner, Appl. Microbiol. 25: 550--552, 1973). A total of 93 organisms isolated on Martin-Lewis media were determined to be Neisseria gonorrhoeae based on the following criteria: presence of gram-negative diplococci, oxidase positivity, and appropriate reaction in the rapid fermentation method. These 93 isolates were then serologically tested with the Phadebact test. The direct method was attempted on the first 46 N. gonorrhoeae isolates. Difficulty in interpreting results was encountered in 39%. Thereafter, the alternate method of boiling was instituted on an additional 47 N. gonorrhoeae isolates, with 2 isolates producing noninterpretable results. All 93 isolates were frozen for a maximum of 2 months in skim milk at -25 degrees C. These isolates were thawed and retyped with the alternate boiling procedure, with 97% being confirmed as N. gonorrhoeae. In addition, 33 Neisseria meningitidis isolates, 14 Neisseria species, and 7 Moraxella species were tested with similar techniques. No positive reactions were observed. A cost effectiveness study using 5, 10, and 20 microliters of the gonococcal reagent was undertaken to reduce the cost of the test. When 10 and 20 microliters of reagent were used, no difficulty was encountered in interpreting the reaction. The coagglutination technique was difficult to read when 5 microliters of reagent was used.
Agglutination Tests↗
Enhancement of coagglutination reactions of the Phadebact gonococcus test by ethylenediaminetetraacetate and ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetate.
Incubation of gonococci under conditions optimal for autolysis resulted in increased sensitivity and enhancement of the coagglutination reaction of the Phadebact gonococcus test. These conditions included an alkaline pH (pH 8.3) and the presence of divalent cation chelators such as ethylenediaminetetraacetic acid or ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid. Heating cell suspensions at 90 degrees C for 15 min before assay by coagglutination produced a further increase in sensitivity and enhancement of the reaction. Gonococcal lipopolysaccharide was found to be an important antigen in these coagglutination reactions. The detection of lipopolysaccharide was markedly enhanced by the addition of chelating agents.
Agglutination↗
Evaluation of the Phadebact Gonococcus Test in the identification of Neisseria gonorrhoeae in a routine diagnostic laboratory.
The Phadebact Gonococcus Test, a coagglutination procedure for the confirmation of identity of presumptive N gonorrhoeae isolates, was evaluated under normal working conditions in a routine diagnostic laboratory and compared with an immunofluorescence technique. Of 166 isolates of N gonorrhoeae from urogenital, rectal, and pharyngeal sites, 164 gave a positive coagglutination reaction, and one of the two negative isolates gave a positive reaction on retesting after subculture. There were no cross-reactions with other organisms tested. This was in contrast with the immunofluorescence technique; with this method, three of 46 isolates of N meningitidis fluoresced brightly, and a further 17 isolates of N meningitidis gave reactions that were difficult to interpret. The coagglutination test is rapid, simple and a more specific alternative to immunofluorescence.
Agglutination Tests↗
Induction of a cell wall variant of the gonococcus by human amylase.
Human amylase has been shown to be an effective and powerful inhibitor of the gonococcus in vitro. Its action appears to be on the bacterial cell wall. When tested in osmotically stabilised liquid media the organism was not inhibited; however the organisms which grew in such cultures appeared to be cell wall variants, which were less sensitive to penicillin than the parent strains. Studies on these variants suggest that they are "transitional-phase variants." Since cervical mucus has much in common with an alkaline osmotically stabilised liquid medium and also contains a high concentration of amylase, it seems possible that these variants may occur in vivo.
Amylases↗
[Gonococcus-associated arthritis].
The various forms of arthritis associated with a gonococcus infection are pathogenetically and clinically differentiated. Whereas an infectious systemic process with different clinical symptoms is said to be underlying the arthritis-dermatitis syndrome as well as the septic GO-arthritis, the third form is para-infectious reactive arthritis. It is often difficult to diagnose an infectious GO-arthritis, as direct evidence of the virus found in joint and blood is rarely positive, so that the diagnosis can be affirmed or negated on the basis of clinical facts of the reaction of arthritis after an appropriate antibiotic therapy. Differential diagnostic considerations may help to find the correct diagnosis in view of an acute urethritis arthritis.
Arthritis, Infectious↗