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Paper strip assay for neisseria species
| Details |
Inventors: Cohenford, Menashi A.;
Assignee:
Primary Examiner: Rosen; Sam
Assistant Examiner:
Attorney, Agent or Firm: Linek; Ernest V., Brown; Donald
The present invention is directed to a rapid enzymatic method using chromogenic substrates for the identification of Neisseria gonorrhoeae, Neisseria meningitidis and Neisseria lactamica. The assay correlated 100% in its identification of pathogenic Neisseria with modified NYC fermentation medium. The assay is more sensitive in its direction of prolylaminopeptidase activity in Neisseria meningitidis than any of the commercially available systems. The test method of the present invention is performed by first applying a small amount of buffer, then applying colonial growth, to each of three test areas (PAP, GAP, and BDG) on filter paper test strips. The strips are then incubated at from about 35.degree.-37.degree. for 10 minutes or at room temperature for 20 minutes. If the BDG area is positive (blue-green color) the isolate is identified as Neisseria lactamica. If the BDG area is negative, a chromogenic reagent, such as dimethylaminocinnaminaldehyde, is added to the PAP and GAP test areas. If a purple color develops in area B. N. meningitidis is present; however, if a red color develops solely in area C the presence of Neisseria gonorrhoeae is indicated. |
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DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention is directed to a test system for the rapid identification of pathogenic Neisseria from plated media growth and to the method of using that system.
The system utilizes three chromogenic substrates, preferably impregnated on filter paper, to detect the preformed enzymes associated with Neisseria gonorrhoeae, Neisseria meningitidis, and Neisseria lactamica.
Referring to the Figures, there are depicted in FIGS.
1, 2 and 3, three views of one preferred arrangement for the test strip of the present invention.
As illustrated, the preferred test strip comprises a section of filter paper 10 and a separator 12.
The separator 12 has three openings 14, 16, and 18, exposing the filter paper 10.
Beneath filter paper 10 is a backing 20, which supports the filter paper.
Openings 14, 16 and 18 serve as the test areas and are each individually treated with an effective amount of an enzyme substrate specific for Neisseria lactamica (e.
g.
, 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside, for BDG), Neisseria meningitidis (e.
g.
, 1-gamma-glutamyl-p-nitroanalide hydrochloride, for GAP), and Neisseria gonorrhoeae (e.
g.
, 1-proline-beta-naphthylamide hydrochloride, for PAP).
The separator 12 prevents interaction among the three test areas.
Suspect colonies are taken from suitable selective media (e.
g.
, Modified Thayer Martin agar) with a wooden applicator stick, swab or inoculating needle and smeared on the three test areas of the filter paper test strip.
With 10-20 minutes the test strip is read for the detection of the enzymes produced by Neisseria as described for example by Yajko et al.
, supra; D'Amato et al.
, supra; and Doyle et al.
, J.
Clin.
Microbiol.
, 3, 383-387 (1984).
Evaluations of 13 strains of Neisseria gonorrhoeae, 10 strains of Neisseria meningitidis, 4 strains of Neisseria lactamica, and 1 strain of Branhamella catarrhalis indicated that the product and method of the present invention correlated 100% with modified NYC fermentation medium
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Fuel Cells 
