Abstract: The present invention discloses a mutant Neisseria having extensive membrane blebbing, both an indicium and a cause of virulence in the gonococcus and meningococcus. Methods are disclosed for making and characterizing the mutant, bmrRS. Methods are disclosed for isolating bmrRS membranes for use as a vaccine. Methods are also disclosed for the use of the mutant for determining the virulence of clinical samples of N. gonorrhoeae and N. meningitidis. Methods are also disclosed for the screening of antibiotics targeted to virulent Neisseria.

Abstract: The present invention relates to a method of transferring at least two saccharide units with a polyglycosyltransferase, a polyglycosyltransferase and a gene encoding such a polyglycosyltransferase.

Abstract: The present invention discloses a mutant Neisseria having extensive membrane blebbing, both an indicium and a cause of virulence in the gonococcus and meningococcus. Methods are disclosed for making and characterizing the mutant, bmrRS. Methods are disclosed for isolating bmrRS membranes for use as a vaccine. Methods are also disclosed for the use of the mutant for determining the virulence of clinical samples of N. gonorrhoeae and N. meningitidis. Methods are also disclosed for the screening of antibiotics targeted to virulent Neisseria.

Abstract: The present invention relates to a method of transferring at least two saccharide units with a polyglycosyltransferase, a polyglycosyltransferase and a gene encoding such a polyglycosyltransferase.

Abstract: The present invention relates to a universal test systems and methods of use thereof for identifying a microorganism among at least two groups of widely divergent microorganisms. The universal test system comprises a predetermined combination of non-redundant biochemical tests comprising a substrate for at least one enzyme wherein the substrate, if acted on by the enzyme results in formation of a detectable product. Detectable products from the combination of biochemical tests are then used to identify the microorganism.

Abstract: The present invention concerns a method and an analytical device for the simultaneous detection of at least two organisms selected from the group consisting of Chlamydia trachomatis (CT), Neisseria gonorrhea (NG), and Mycoplasma (M) from a single specimen.

Abstract: Compositions comprising outer membrane protein "CD", and peptides and oligopeptides thereof, of Branhamella catarrhalis are described. Additionally, nucleotide sequences encoding the protein, peptide or oligopeptide are disclosed, as well as recombinant vectors containing these sequences. Protein, peptide or oligopeptide can be produced from host cell systems containing these recombinant vectors. Peptides and oligopeptides can also be chemically synthesized. Disclosed are the uses of the protein, peptides and oligopeptides as antigens for vaccine formulations, and as antigens in diagnostic immunoassays. The nucleotide sequences are useful for constructing vectors for use as vaccines for insertion into attenuated bacteria in constructing a recombinant bacterial vaccine, and for inserting into a viral vector in constructing a recombinant viral vaccine. Also described is the use of nucleotide sequences related to the gene encoding CD as primers and/or probes in molecular diagnostic assays for the detection of B.

Abstract: The present invention relates to the lower molecular weight subunit of the human transferrin receptor of a strain of N. meningitidis, in purified form, as well as to a vaccinal pharmaceutical composition intended for the prevention or attenuation of the effects of an N. meningitidis infection, containing the said subunit in purified form.

Abstract: A vaccinal pharmaceutical composition which comprises, as therapeutic agents, at least a first and a second molecule capable of binding to human transferrin; the said first molecule originating from a first strain of N. meningitidis which possesses a human transferrin receptor in which the lower molecular weight subunit (Tbp2) is recognised by an antiserum to the receptor of N. meningitidis strain 2394 (receptor 2394) and is not recognised by an antiserum to the receptor of N. meningitidis strain 2169 (receptor 2169); and at least a second molecule originating from a second strain of N. meningitidis which possesses a human transferrin receptor in which the lower molecular weight subunit (Tbp2) is recognised by an anti-receptor 2169 antiserum and is not recognised by an anti-receptor 2394 antiserum.

Abstract: Compositions comprising outer membrane protein "CD", and peptides thereof, of Branhamella catarrhalis are described. Additionally, nucleotide sequences encoding the protein or peptide are disclosed, as well as recombinant vectors containing these sequences. Protein or peptide can be produced from host cell systems containing these recombinant vectors. Peptides can also be chemically synthesized. Disclosed are the uses of the protein and peptides as antigens for vaccine formulations, and as antigens in diagnostic immunoassays. The nucleotide sequences are useful for inserting into a viral vector in constructing a recombinant viral vaccine. Also described is the use of nucleotide sequences related to the gene encoding CD as primers and/or probes in molecular diagnostic assays for the detection of B. catarrhalis.

Abstract: A nucleotide sequence characteristic of Neisseria gonorrhoeae is disclosed. The sequence can be the basis for hybridization type, nucleic acid-based, rapid, in vitro diagnostic assays. The unique nature of the sequence makes it possible to clearly discriminate N. gonorrhoeae from other Neisseria species thus eliminating or substantially reducing the number of false positive readings. A 350 base pair N. gonorrhoeae DNA restriction fragment was cloned after subtractive hybridization to Neisseria meningitidis DNA. In further cloning experiments the sequences adjacent to the original 350 base pair fragment were determined. A portion of this sequence was shown to detect 105 of 106 N. gonorrhoeae strains and no other Neisseria species. In addition to use as detection probes, all or portions of the nucleotide sequence can be used as a ligand for the sandwich capture of N. gonorrhoeae sequences and as primers for in vitro amplification of N. gonorrhoeae sequences.

Abstract: An extract based on modified bacterial proteins includes a mixture of acid bacterial polyanions having a molecular weight in the range from 10,000 to 1,000,000 and an isoelectric point in the range from 2.5 to 5.5, and in which the added weights of the constituent amino acids amount to at least 50% of the extract. The preparation or this protein extract includes a cultivation of bacteria in an aqueous medium and then the alkaline extraction of this bacterial suspension and the purification of the protein extract. The alkaline extraction is carried out in the presence of a dilute aqueous source of OH.sup.- ions and at a stable pH in the range from 11 to 13, the decrease of this pH during the acid extraction not exceeding 0.4. The protein extract thus obtained can be used as an active ingredient in a pharmaceutical composition.

Abstract: Enhanced extraction of solubilized antigens is obtained from bacteria such as Chlamydia and Neisseri by the use of a buffer containing a zwitterionic surface active agent, especially CHAPS or CHAPSO, in the absence of divalent cations. The extraction is conducted at elevated temperature, and provides a sample useful in assays for the presence of the bacteria.

Abstract: A novel colorimetric method for beta-lactamase assay and its applications are disclosed. The novel assay method for beta-lactamase is based on the discovery described in this invention. The chromophore formed by oxidation of either the N-alkyl derivative of p-phenylenediamine or the 3,3',5,5'-tetraalkyl derivative of benzidine can be decolorized by the open beta-lactam ring end product resulting from the hydrolysis of a penicillin in the presence of a mercury-containing compound. The same chromophore can be decolorized by the open beta-lactam ring end product resulting from the hydrolysis of a cephalosporin in either the presence or the absence of the mercury-containing compound. None of the intact beta-lactam antibiotics can decolorize the chromophore in either the presence or the absence of the mercury-containing compound. The final concentration of the mercury-containing compound in the decolorization mixture ranges from approximately 0.

Abstract: A nucleotide sequence characteristic of Neisseria gonorrhoeae is disclosed. The sequence can be the basis for hybridization type, nucleic acid-based, rapid, in vitro diagnostic assays. The unique nature of the sequence makes it possible to clearly discriminate N. gonorrhoeae from other Neisseria species thus eliminating or substantially reducing the number of false positive readings. A 350 base pair N. gonorrhoeae DNA restriction fragment was cloned after subtractive hybridization to Neisseria meningitidis DNA. In further cloning experiments the sequences adjacent to the original 350 base pair fragment were determined. A portion of this sequence was shown to detect 105 of 106 N. gonorrhoeae strains and no other Neisseria species. In addition to use as detection probes, all or portions of the nucleotide sequence can be used as a ligand for the sandwich capture of N. gonorrhoeae sequences and as primers for in vitro amplification of N. gonorrhoeae sequences.

Abstract: A Neisseria-specific DNA probe is chosen from the group of the oligonucleotides Opa 1, Opa 2, Opa 3, P III, Pil 1, IgA 1 and IgA 2 or from sequences complementary to these and it can have up to 5 further nucleotides at the 5' and/or 3' end. The probe is suitable for the detection of at least one of the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis under hybridization conditions by the detection of the hybrid formation.

Abstract: The present invention is directed to recombinant hosts which contain and express various Type II restriction endonuclease and/or modification methylase genes. In particular, the present invention is concerned with the cloned restriction endonucleases, NgoAIII and NgoAI, which recognize and cleave within or near the double-stranded DNA sequence, 5' CCGCGG 3' and 5' PuGCGCPy 3', respectively. Also provided in this invention are cloned modification methylase genes corresponding to said restriction endonucleases. This invention is further concerned with a cloned modification methylase, M.NgoAII. One source of these enzymes is Neisseria gonorrhoeae, although other microorganisms may be used to isolate the restriction endonuclease isoschizomers and modification methylase isoschizomers of this invention.

Abstract: An extraction composition is useful for lysing chlamydial, gonococcal or herpes organisms and extracting detectable antigen from the organisms. In particular, this composition has a high pH (at least about 8) and comprises an alcoholamine which is effective in extracting antigen. It is particularly useful for extracting either or both of the lipopolysaccharide and major outer membrane protein chlamydial antigens.

Abstract: The invention discloses nucleic acid probes specific for Neisseria gonorrhoea useful in diagnostic methods to detect the presence of N. gonorrhoea in biological specimens.

Abstract: The invention discloses nucleic acid probes specific for Neisseria gonorrhoea useful in diagnostic methods to detect the presence of N. gonorrhoea in biological specimens.

Abstract: A buffered composition includes whole chlamydial or gonococcal organisms or an antigen thereof, and a nonionic fluorinated surfactant in an amount of at least about 0.05 weigh percent. This composition is useful in a positive control test used in diagnostic assays for chlamydial or gonococcal organisms. It is particularly useful when coated in an article used in such assays, such as disposable test devices having nonionic polyamide filtration membranes in the bottom of test wells.

Abstract: A specimen containing N. lactamica, N. meningitidis, N. gonorrhoeae or B. catarrhalis is incubated simultaneously with a first substrate for beta-galactosidase and a second substrate for gamma-glutamyl aminopeptidase to form reaction products with one or the other Neisseriae which yields a detectible first or second signal distinct from each other depending upon the presence of the enzyme. Signals may comprise distinct colors which emit either in the absence or presence of diazo dye coupler. A third substrate specific for prolyliminopeptidase in N. gonorrhoeae may be added to form a detectible third signal of the same type. The third substrate may be incubated simultaneously with the first two substrates or later. The absence of all of the first three signals is a positive indication that the specimen contains B. catarrhalis. Also, a diagnostic test kit for performing the above tests with or without lectin or antibody for agglutination based upon specific recognition.

Abstract: The invention discloses nucleic acid probes specific for Neisseria gonorrhoea useful in diagnostic methods to detect the presence of N. gonorrhoea in biological specimens.

Abstract: Antigens from chlamydial or gonococcal organisms in specimens containing whole blood, mucus or components thereof can be rapidly and sensitively determined using a polyamide microporous membrane which is coated with a surfactant. This determination is accomplished by contacting extracted antigen with the coated polyamide microporous membrane which is substantially free of any particulate matter. The membrane has an average pore size of from about 1 to about 10 .mu.meter. Within about 10 minutes of that contacting, antigen bound to the coated membrane is contacted with chlamydial or gonococcal antibody, respectively, so as to form an immunological complex on the membrane. The presence of the complex on the membrane is then determined as a measure of the amount of chlamydial or gonococcal antigen present in the specimen.

Abstract: Methods are disclosed for detecting an antigen in a biological sample. The methods involve providing in combination a solid support, which is substantially free of specific binding proteins, and a medium comprising an antigen from the sample and an antibody for the antigen. The combination is incubated under conditions sufficient for the antibody when bound to the antigen to bind to the support. The presence or amount of antibody on the support or in the medium is determined and is related to the presence of antigen in the sample. The methods have particular application to the detection of gram-negative bacteria.

Abstract: This invention relates to specific discrete nucleotide sequences. More precisely, this invention relates to a composition comprising discrete nucleotide sequences that are specific for Neisseria gonorrhoeae, as defined hereinbelow, i.e. the compositions of the subject invention can be utilized to detect Neisseria gonorrhoeae chromosomal DNA and thus, Neisseria gonorrhoeae.

Abstract: A selective growth medium for the isolation of Mobiluncus includes a medium base including an aqueous preparation of a gelling agent and a nutrient broth effective for culturing Mobiluncus. The selective growth medium of the invention further includes a first antimicrobial agent in effective antimicrobial amounts, the antimicrobial agent being selected from the group consisting of colistin, nalidixic acid, and combinations of colistin and nalidixic acid, the colistin having a concentration less than 32 micrograms/ml and the nalidixic acid having a concentration less than 100 micrograms/ml. Tinidazole is included as a second antimicrobial agent in effective, antimicrobial amounts not greater than 1.0 micrograms/ml. Nile Blue A is also included in Gardnerella Vaginalis inhibiting concentrations.

Abstract: A nutrient growth medium which is highly selective for pathogenic Neisseria is provided. The nutrient growth medium includes an antibiotic agent which is a combination of vancomycin and lincomycin. The combination of lincomycin and vancomycin allows the use of lower concentrations of vancomycin than are generally employed in culture media used for the selective isolation of pathogenic Neisseria species.

Abstract: A bacterial lectin isolatable from Neisseria gonorrhoeae is disclosed. This lectin binds to gonococcal carbohydrates such as gangliotetraosylceramide, has a relative molecular weight of about 22,400 daltons, and an isolectric pH value in the range of about 6.1 to about 6.4. The disclosed lectin is useful as a constituent of a vaccine against gonorrhea and as a diagnostic means for gonorrhea. A method for isolating this lectin is also disclosed, as well as means for producing it.

Abstract: Monoclonal antibodies specific to Neisseria gonorrhoeae lipopolysaccharide components having no cross-reactivity with N. meningitidis have been produced and found useful in the diagnosis of the presence of N. gonorrhoeae.

Abstract: Process for preparing a detoxified polysaccharide-outer membrane protein complex from bacterial envelopes; the so-obtained products which are useful as vaccines against infection by the same bacteria and method for protecting animals against the same infection by administration of a pharmaceutical composition containing the detoxified polysaccharide-outer membrane protein complexes.

Abstract: There are provided a crystalline and single rod products derivable from the Pili of Type 1 and Type 2 Neisseria gonorrhoese organisms. There are provided methods of growing said organisms to produce the maximum yield of Pili and procedures for purifying said Pili to produce said crystalline material. There are further provided methods of utilizing said Pili to determine the presence, in a system infectable by N. gonorrhoese organisms, of antibodies to the Pili of said organisms, and methods of serotyping said Pili. There is also provided a mode of utilizing said crystalline material to provide a substantial degree of immunization infection by N. gonorrhoese in mammalian systems susceptible to such infection.

Abstract: A method for identification of a microorganism includes combining in a suitable fluid the microorganism and a latex containing a particle coated with about 1 to 150 .mu.g per ml of latex of a lectin specific for the microorganism. Binding of the microorganism and lectin causes agglutination which is detected. The microorganism is identified by the agglutination as that organism which is specific for the lectin. The invention includes a reagent which may be included with other materials in a kit of materials useful for performing the method of the invention.

Abstract: The method disclosed herein allows for the pretreatment of organisms, suspected of being a material of interest, prior to performing an assay for a determination thereof. The method comprises contacting the organism in an aqueous medium with a composition comprising (1) an enzyme capable of hydrolyzing bonds between N-acetylglucosamine and N-acetylmuramic acid and (2) a chelating agent, in amounts and under conditions sufficient to produce a homogeneous suspension of the organism of interest in the aqueous medium but insufficient to produce lysed cells or spheroplasts. The method and composition are particularly applicable to the pretreatment of cells of gram negative bacteria such as, for example, N. gonorrhoeae, and of chlamydia.

Abstract: A medicament and method for inducing immunity in to infectious bovine keratoconjunctivitis in cattle. The medicament comprises the gram negative cocci Neisseria or Branhamella which are non-etiological agents of infectious Keratoconjunctivitis yet unexpectedly are found to afford an immunity to infectious bovine keratoconjunctivitis when administered to cattle.

Abstract: Immunoassay detection of bacterial diseases, bacteria, and microorganisms using a deionized water collection medium. The preferred enzyme immunoassay is of particular use in clinical or home testing application for detection of bacteria such as gonococcus, antigens derived from such bacteria, and antibodies against the bacteria. A colorimetric detection technique may be employed using chromogenic solutions containing tetramethylbenzidine or water soluble derivatives of tetramethylbenzidine.

Abstract: The present disclosure relates to a solid phase immunoassay for the detection of Neisseria gonorrhoeae antigens in a clinical specimen, wherein the Neisseria gonorrhoeae antigens to be determined are coated or adsorbed on the solid phase.

Abstract: There are provided a crystalline and single rod products derivable from the Pili of Type 1 and Type 2 Neisseria gonorrhoeae organisms. There are provided methods of growing said organisms to produce the maximum yield of Pili and procedures for purifying said Pili to produce said crystalline material. There are further provided methods of utilizing said Pili to determine the presence, in a system infectable by N. gonorrhoeae organisms, of antibodies to the Pili of said organisms, and methods of serotyping said Pili. There is also provided a mode of utilizing said crystalline material to provide a substantial degree of immunization to infection by N. gonorrhoeae in mammalian systems susceptible to such infection.

Abstract: A strain of Neisseria gonorrhoeae ATCC 31953 is described which is abnormal in that it has characteristically poor growth on chocolate agar at a temperature range of about 30.degree. C. to about 37.degree. C. in a CO.sub.2 atmosphere suitable for growth of N. gonorrhoeae. This strain is resistant to nalidixic acid at the 5-10 mcg/ml level and resistant to streptomycin at the 1000 mcg/ml level or greater. N. gonorrhoeae ATCC 31953 is a test strain suitable for use in the method described for the laboratory diagnosis of gonorrhea.

Abstract: Method for rapid, high intensity visual indication of the presence of Neisseria bacteria species utilizing a new and improved peptone buffer and pH indicator reagent system as the specimen carrier for oxidation series testing, which series test may also include beta lactamase enzyme detection utilizing the reaction of the same reagent system carrying the specimen with a penicillin substrate.

Abstract: An over the counter swab kit for self detection of gonorrhea in the male using saline ampul. A method wherein a pledget in the form of fibrous material is impregnated with a compound which reacts with Neisseria gonorrhea to produce a color change. The pledget, before the test, is in a dry condition and is activated by a wetting agent such as saline, when placed in contact with the pledget. The chemical compound used and incorporated into the pledget is selected from a group consisting of phenylenediamines. A sufficient amount of saline is held within a frangible ampul to wet the pledget and then to be drawn by capillary action through the swab on which a sample of exudate as a specimen is placed. The chemical compound is in the form of a mineral acid salt which dissolves in the saline and is transported by capillary action through the pledget into the swab. A purple color is created on the tip of the swab at the site of the bacteria and is a positive test for gonorrhea.

Abstract: An over the counter swab kit for self detection of gonorrhea in the male using an ampul containing 1% N, N, N' N' tetramethyl-p- phenylenediamene or tetramethyl chromogen. The method of taking a sample is also shown in which the open end of a tube is filled with a plug in the form of fibrous material. After a sample is taken of exudate from a male, the 1% solution of tetramethyl chromogen reacts with Neisseria gonorrhea which may be present in the exudate to produce a color change. The plug, before the test, is in a dry condition, the plug is activated only by the tetramethyl chromogen which is placed below the plug in the tube and the ampul broken to release its contents. A sufficient amount of 1% tetramethyl chromogen is held within the frangible ampul to wet the plug and this amount is 1/2 the volume than is used in the saline ampul in my companion patent application entitled Over the Counter Swab Kit for Self Detection of Gonorrhea in the Male Using Saline Ampul.

Abstract: A procedure for detection of Neisseria gonorrhoeae in a microbial growth culture. Lysates of the suspect organism are tested by reverse passive hemagglutination with particles sensitized with antibodies to eleven N. gonorrhoeae strains, ATCC 31397, 31398, 31399, 31400, 31401, 31402, 31403, 31404, 31405, 31406, and 31407.

Abstract: Gonococcal antigens are solubilized by a process involving trypsin-digestion.

Abstract: A method for the continuous culturing of microbes in a plug-flow reactor which comprises the steps of:A. supplying medium to microbes immobilized on a porous inorganic support at a rate sufficient to maintain such microbes substantially in a logarithmic growth stateandB. removing microbe-containing effluent from the immobilized microbes at a rate equal to the medium supply rate, wherein the microbes are selected from the group consisting of bacteria, yeasts, and fungus-like organisms; such reactor is operated continuously in a substantially plug-flow mode; the immobilized microbes are substantially covered by said medium; and such porous inorganic support has a controlled porosity such that at least 70% of the pores, on a pore size distribution basis, have a pore diameter,a. in the case of bacteria, at least as large as the smallest major dimension of the microbes but less than about five times the largest major dimension of the microbes;b.

Abstract: A microbial growth medium for the isolation and rapid detection of Neisseria gonorrhoeae and Neisseria meningitidis, having incorporated therein an iron-dextran complex in a quantity sufficient to stimulate maximum colony growth.

Abstract: The disclosure is directed to a method, reagents and apparatus for the rapid (one to four hours) identification of Neisseria gonorrhoeae and Neisseria meningitidis from cultures grown on a selective or a non-selective medium. One drop of growth suspended in sterile salt solution is placed in each of a plurality of miniature reaction chambers supported on a common base and incubated at 35.degree.-37.degree. C. for one to four hours. A dried substrate and buffer is contained in each chamber. A detector reagent such as a diazo dye in an aqueous solution with a polar solvent is added to each chamber after incubation. The color change in each chamber is noted and compared with a profile to identify N. gonorrhoeae and Neisseria meningitidis. Synthetic substrates rather than sugars are used in the reaction chambers. The substrates are naphthyl derivatives, .beta.-naphthyl or .beta.-naphthylamides, and are(1) .beta.-naphthyl-.beta.,D-galactopyranoside(2) N-L-.lambda.-glutamyl-.beta.

Abstract: Bacteria of the genus Neisseria can be detected in a sample by testing for the presence of an enzyme capable of oxidizing 1,2-propanediol and reducing NAD (nicotinamide-adenine-dinucleotide). The complete structure of the enzyme is not known but, because of those two characterizing properties, the nomenclature 1,2-propanediol dehydrogenase therefor is proposed herein.