DK142823B - Method and Instrument for Rapid Examination of a Male Patient for Neisseria gonorrheae. - Google Patents

Description

142823 i

The present invention relates to a method and instrument for rapid examination of a male patient for Neisseria gonorrheae without culture and without gram staining of this organism.

From Acta Universitatis Carolinae Medica, Vol. 27, No. 1/2, pages 47-50 (1971), a test method for Neisseria gonorrheae (Ng) known without prior cultivation for the growth of the organisms is known and used a combination of two different reagents, p-phenylenediaminosulfate and α-naphthol, which are dissolved in Chelaton I in solution to obtain a solution which, after adjusting to pH 3.5-4.5, is used to impregnate filter paper which then after drying is used for direct diagnosis of urethral secretion for Neisseria gonorrheae.

The sample described in said journal has been known since 1956 and is referred to in British J. Venereal Diseases 43 (1967), page 73, wherein the sample using dimethyl = phenylenediamine plus α-naphthol as reagents is described as resulting in a light blue color as resulting from the formation of a blue indophenol dye under the prevailing pH coupling conditions.

According to the latter literature site, the test method requires the use of an ample amount of urethra secretion which must be sufficient for application on both sides of the test paper and the test method result can be read after 10 minutes. However, it has been found that more than 10% of the samples lead to incorrect positive test results and that 9% of the samples lead to incorrect negative test results.

Although the test method known from said literature sites is faster than certain other known test methods, it is desirable to have a considerably faster test method, but in particular it is desirable to have a method which is considerably more reliable than the known one.

142823 2

The process of the invention is characterized by taking a sample of the patient's urethral fluid directly onto a sterile swab, contacting the swab with a porous absorbent tampon which has been impregnated with a single color forming reagent selected from p-aminodimethyl = aniline oxalate, N , N-dimethyl-p-phenylenediamine dihydrochloride, N, N-dimethyl-p-phenylenediamine oxalate, N, N-dimethyl-p-phenylene = diamine monohydrochloride and Ν, Ν, Ν ', N'-tetramethyl-p-phenylene = diamine dihydrochloride, dried tampon is placed in a transparent plastic tube with flexible sidewalls and a closed end in which a fragile ampoule containing physiological saline solution or water of pH 6.5-7.2 is placed in the tube as solvent for contacting the tampon. the reagent in the tampon, which ampoule is then broken by compression of the flexible side walls near the closed end of the tube, after which the sample wetted on the swab with the reagent dissolved in the contents of the ampoule is observed. 30 to 120 seconds to detect any staining indicating a positive reaction.

This makes it easy to make a quick diagnosis for Neisseria gonorrheae directly on the secreted urethral secretion, whereby within two minutes a test result can be obtained which shows 95-98% consistency with the result obtained by gram staining in connection with a bacteriological test method.

The diagnostic instrument for carrying out the method according to the invention is characterized in that it comprises a sterile cotton swab and a transparent end closure tube with flexible sidewalls designed for insertion of the cotton swab and in contact with one another. placed partly a porous absorbent tampon which has been impregnated with a single color forming reagent selected from p-aminodimethylaniline oxalate, N, N-dimethyl-p-phenylenediamine hydrochloride, N, N-dimethyl-p-phenylenediamine oxalate, N, N-di = methyl-p-phenylenediamine monohydrochloride and Ν, Ν, Ν ', N'-tetra = methyl-p-phenylenediamine dihydrochloride, and dried, partly a fragile ampoule containing physiological saline solution or water of pH 6.5-7.2 closest to the tube. closed end.

The tampon may be made of any suitable material, such as dacron fibers, cotton fibers, or other porous material, and impregnated or saturated with one of the said color-forming reagents under certain conditions hereinafter referred to and then dried. It remains in the dry state until activated by the wetting agent contained in the vial.

The tampon is the most important component of the diagnostic instrument as it contains the reactive chemical which is able to identify the gonococcus. In the dry state, the tampon made in this way will remain stable and has great durability. Another component contained in the instrument is the wetting agent, which is separated from the tampon by virtue of being contained in a brittle ampoule. When the brittle ampoule is broken, the wetting agent contained will release and activate the reagent in the tampon which, after being made very sensitive, will contact the sample at the tip of the collection swab. The activated reagent reacts with the gonococci present and causes the sample on the swab to get a characteristic color depending on the choice of reagent used. The tampon, which is almost colorless, does not cause a disruptive reaction because it does not react colorimetrically. The color change takes place on the sample collected on the swab. The reaction time needed to indicate a positive sample is usually from 30 to 120 seconds.

The wetting agent contained in the vial may consist of water, physiological saline solution, pH buffer solutions or any other solution or combination of solutions which enhance or sensitize the oxidase reaction. The pH value of the wetting agent is desirable to prevent auto-oxidation of the reagent in the tampon. The preferred pH range is from 6.5 to 7.2.

142823 4

The invention will be explained in more detail below with reference to the drawing, in which fig. 1 shows an instrument suitable for use in carrying out the method according to the invention, the instrument being shown before being used for a test and in the form in which it is sold as a two-part packaged system; and FIG. 2 shows the embodiment of FIG. 1 after it has been used to perform a test.

With reference to the drawing, which shows an embodiment of the present instrument for carrying out the process according to the invention, 10 represents a tube of transparent, flexible plastic into which is inserted a cotton tampon 12 which has been impregnated or saturated with the reagent and dried . The tampon is positioned next to a brittle glass vial 14 located at the closed end of the tube and a lid 16 is placed on the open end of the tube. A sterile cotton swab 18 with a plastic grip pin 20 for use in sampling exudate is enclosed in a separate sterile wrap 22 and the tube and wrap are packaged together in a casing or other container not shown. Although the wall of the plastic tube 10 is flexible, it is sufficiently strong for the tube to normally maintain a cylindrical shape and to allow easy insertion of the swab after an exudate sample is taken.

The ampoule contains a suitable wetting agent as described above and is sufficiently brittle to be easily broken when the sides of the flexible tube are pressed inward between the thumb and forefinger, thereby breaking the ampoule and allowing its liquid content to moisten the material. with which the tampon is impregnated, the tampon being normally pressed into the liquid and the glass pieces as the cotton swab is inserted into the tube. When the swab containing the sample contacts the tampon, the reaction between the swab swab and the reagent in the tampon begins immediately, and if gonococci are present, the swab swab color changes, usually to violet, red-orange or dark gray, depending on the reagent used. the tampon so that a positive sample is indicated. The reaction time for indicating a positive sample is usually from 30 to 120 seconds.

A preferred process for preparing the tampon and for preparing the wetting agent is as follows:

A 1% solution of Ν, Ν, Ν1, N'-tetramethyl-p-phenylenediamine dihydrochloride is prepared as follows: 1 g of the reagent is added to 35 ml of distilled water which has been brought to a boil and allowed to cool to room temperature. Resolution is done by rapid stirring.

64 ml of ethyl alcohol (Fisher A-407) is then added and stirred. Cotton Webril strips, 2 cm wide and 50 cm long, are dipped one at a time in the solution, placed on a nylon net in a horizontal position and allowed to air dry. The dry strips are then cut into 1 1/2 cm long pieces. These are used as tampons.

A physiological saline solution is prepared by adding 0.85 g of sodium chloride to 99 ml of distilled water and adjusting the pH to 7.0. The solution is used as a wetting agent. Fragile ampoules containing 1/2 ml of this solution are prepared, this amount being required for sufficient wetting of the 2 cm x 1.5 cm tampon.

The ampoule, approximately 7 mm in diameter and 35 mm long, is placed at the bottom of a flexible plastic tube having an internal diameter of approx. 8 mm and a length of 160 mm. A tampon prepared as above is rolled up and pressed down into the tube so that it is in contact with the ampoule. A telescopic lid with a length of approx. Insert 65 mm on the open end of the plastic tube to close it.

A sterile "dacron" cotton ball placed on the end of an approx. 150 mm long plastic stick is packaged with the reagent system and this combination constitutes the diagnostic instrument. The swab is separately contained in its own sterile parchment wrap.

The above two parts, which are part of a single package, can be sold to doctors, clinics and hospitals via normal pharmaceutical channels, and the package is immediately available for examination and rapid diagnosis of the patient's condition at the point where a sample is to be taken. When a gonorrhea diagnostic examination is performed, the gasket is opened and the sterile swab is removed from its cover and a sample of the exudate is collected at the tip of the swab. The telescopic lid is then removed from the plastic tube and the swab inserted with the tip first into the plastic tube. Replace the telescopic lid. The flexible tube is then compressed at the vial portion, thereby breaking the vial and delivering its contents. At the same time, the telescopic lid is pushed down, whereby the swab with the sample comes into contact with the tampon. This depression also depresses the tampon into the wetting agent, thereby activating the reagent. When the sample on the swab becomes violet in two minutes, the sample for gonorrheae is positive. The swab can be removed a few centimeters from the tampon to facilitate reading of the result, ie. it changed the color of the sample on the swab. The use of this test method therefore provides the physician or clinician with a quick and accurate diagnostic tool at the first stage of his differential diagnosis.

To further elucidate the sensitivity of the test method and its utility in detecting the gonococcal bacterium exactly at the site where the sample is collected on the swab, the above procedure is repeated and a culture of neisseria gonorrhea is used as the representative of the exudate in the previous example. In this example, the swab is used to extract a few colonies from a petri dish culture. The same procedure is performed with the instrument. A clear violet staining of the bacteria on the swab will be observed within one minute. The almost colorless tampon and "dacron" cotton swab

Claims (2)

1A2823 white background makes the identification of the organisms on the swab clear and clear, as is the case when the exudate is taken directly from the patient. According to the U.S.A. Patent No. 3,450,129 has previously used brittle ampoules to hold reagents, but the method known from that patent is specifically intended to transport a sample to a laboratory and to maintain the viability of the bacteria until it can be cultured. The method according to the invention, on the other hand, is a diagnostic method which does not require cultivation. This is a direct investigation method. The diagnosis is made at the time of sampling. Claims. A method for rapid examination of a male patient for Neisseria gonorrheae without culture and without gram staining of this organism, characterized by taking a sample of the patient's urethral secretion directly on a sterile swab (18), contacting the swab with a porous, absorbent tampon (12) which has been impregnated with a single color forming reagent selected from p-aminodimethylaniline oxalate, N, N-dimethyl-p-phenylenediamine = dihydrochloride, N, N-dimethyl-p-phenylenediamine oxalate, N, N-dimethyl-p -phenylenediamine monohydrochloride and Ν, Ν, Ν ', Ν'-tetramethyl-p-phenylenediamine dihydrochloride, which is dried in a tampon in a transparent plastic tube (10) with flexible sidewalls and a closed end, in contact with the tampon. placed a fragile ampoule (14) containing physiological saline solution or water of pH 6.5-7.2 in the tube as solvent for the reagent in the tampon, which ampoule is then broken by compressing the flexible sidewalls near by the closed end of the tube, after which the sample wetted with the reagent dissolved in the vial contents of the ampoule is observed for 30 to 120 seconds to determine