US3699003A - Diagnostic preparation for the identification of beta-hemolytic streptococci - Google Patents

Diagnostic preparation for the identification of beta-hemolytic streptococci Download PDF

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Publication number
US3699003A
US3699003A US54277A US3699003DA US3699003A US 3699003 A US3699003 A US 3699003A US 54277 A US54277 A US 54277A US 3699003D A US3699003D A US 3699003DA US 3699003 A US3699003 A US 3699003A
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United States
Prior art keywords
reagent
streptococci
esculin
area
salicin
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Expired - Lifetime
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US54277A
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English (en)
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Donald P Kronish
Lee S Zuriff
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Warner Lambert Co LLC
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Warner Lambert Co LLC
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
    • G01N31/22—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using chemical indicators
    • G01N31/221—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using chemical indicators for investigating pH value
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/805—Test papers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/822—Microorganisms using bacteria or actinomycetales
    • Y10S435/885—Streptococcus

Definitions

  • a preparation for the biochemical identification of phemolytic streptococci into serologic groups, e.g., groups A, B, C, D, etc., is disclosed.
  • This preparation comprises a carrier, e.g., a bibulous strip containing a glycerol reagent band, a salicin reagent band and an esculin reagent band.
  • glycerol and salicin reagent bands there are also pH indicator systems.
  • the esculin band contains a compound which reacts with esculin breakdown products to produce a color change. These bands are separated from each other by hydrophobic barriers.
  • a quantity of the beta hemolytic streptococcus whose serological identification is to be determined is transferred onto each of the three reagent bands.
  • a drop of saline or water is added to the salicin area and to the esculin area.
  • the strip is placed in a tube containing 0.3 ml. of saline or water to wet the glycerol band.
  • the reagent impregnated strip is incubated at approximately 37 C. for two to four hours.
  • Group A streptococci which cause most of the clinical manifestations such as rheumatic fever and nephritis, are esculin negative, glycerin negative and sa'licin positive.
  • Group B and C streptococci may be involved in diseases but more often are part of a normal flora in a primary isolation blood plate showing fl-hemolysis. It is important to differentiate between these groups and Group A.
  • Group D streptococci known as enterococci, are often associated with urinary tract infection and rapid identification of these organisms is also clinically important.
  • Streptococci are gram positive organisms, spherical or oval in shape and have a tendency to grow in chains. Those streptococci which possess primary pathogenicity for man and animals belong to the subdivision of hemolytic streptococci. This classification is based on the observation that these streptococci cause lysis of red blood cells, a property which is useful in the initial identification of these organisms in culture material. In a blood plate showing a clear zone resulting from lysis of the red blood cells, the organism is defined as B-hemolytic. Other types of hemolysis have also been described.
  • the preparation of this invention is a carrier, typically a strip of a bibulous material which contains a plurality of distinct areas.
  • Area 1 is impregnated with glycerol, a pH indicator system, and optionally containing nutrient medium and having a pH on the alkaline side of the pH indicator used.
  • Area 2 is a hydrophobic barrier which is capable of preventing the migration of the ingredients. Any substance which will form a waterproof barrier can be advantageously employed. Suitable materials include, for example, waxes, lacquers and a colorless acrylic resin known as Krylon Crystal Clear: this Krylon material is particularly preferred. It is supplied in a hydrocarbon vehicle and may be diluted for ease of application with toluene or other hydrocarbon solvents, for example, methyl, ethyl or isopropyl alcohol.
  • Area 3 is impregnated with salicin, a pH indicator system and optionally containing a nutrient medium and having a pH on the alkaline side of the pH indicator used.
  • Area 4 is a hydrophobic barrier which is the same as previously described for area 2.
  • Area 5 is impregnated with esculin and a ferric salt, for example, ferric chloride, ferric ammonium citrate, ferric nitrate, ferric citrate, or an aluminum salt, for example, aluminum ammonium sulfate, or a lead salt for example, lead acetate or lead nitrate.
  • a ferric salt for example, ferric chloride, ferric ammonium citrate, ferric nitrate, ferric citrate, or an aluminum salt, for example, aluminum ammonium sulfate, or a lead salt for example, lead acetate or lead nitrate.
  • Area 6 is a hydrophobic barrier which is the same as previously described for area 2.
  • Area 7 is optional and can be impregnated with any hydrophobic material as previously described for area 2 and any appropriate dye serving as an identification means.
  • sequence of reagent areas on the strip is not critical as long as each area is separated by a hydrophobic barrier but the configuration described is preferred.
  • the indicator system for areas -1 and 3 are those indicators which will give a color change between about pH 9 to about pH 5.
  • these indicators are chlorophenol red, bromocresol purple, bromophenol red, bromothymol blue, neutral red, u-naphtholphthalein and phenol red.
  • nutrient media are those which are traditionally used in bacterial culture, for example, peptone, tryptone, proteose, tryptose, yeast extract, etc.
  • Bibulous materials which can be employed as the carrier are, for example, those materials which by means of a capillary action can draw a liquid upwards; or materials such as filter paper, felt, porous, ceramic strips, woven or matted glass fiber and the like are suitable.
  • the active ingredients can be impregnated on any absorbent material and then mounted on a suitable support, for example, a plastic strip.
  • a loopful of organism showing fl-hemolysis on a blood agar plate is rubbed into the bottom of area 1 and into areas 3 and 5.
  • a drop of saline or water is also added to areas 3 and 5.
  • the resulting system is then immersed in a 13 x 100 mm. or similar size test tube containing 0.3 ml. of saline and incubated at 37 C. for about 2 to 4 hours.
  • a positive reaction in areas 1 and 3 when phenol red is the indicator system is indicated by the development of a yellow color; a negative color is indicated by the persistence of a red color.
  • the positive reaction is as a result of the fermentation of the sugars and the resulting acidity.
  • a positive reaction in area is indicated by the development of a gray-black color, whereas a negative reaction is indicated by the absence of color change.
  • a positive reaction is the result of the fermentation of esculin and the reaction of hydrolysis products with ferric salts or other color producing compounds.
  • the initial step is to prepare the barrier areas 2 and 4 which act to prevent the migration of the reagent solutions which are subsequently supplied.
  • These barrier zones are applied to suitable carrier, preferably Eaton-Dikeman No. 623 filter paper, by employing a solution of a lacquer which produces a water impervious barrier. After allowing the lacquer solution to dry, the glycerin reagent solution, the salicin reagent solution, and the esculin reagent solution prepared according to the above description is then applied to areas 1, 3 and 5. The areas thus formed are allowed to dry; the filter paper is then cut into strips.
  • One or two loopfuls of streptococci showing ,B-hemolysis on a blood agar plate is rubbed on the bottom of area 1 and onto areas 3 and 5 respectively.
  • the test system thus produced is added to a 13 x mm. tube containing 0.3 ml. of saline so that area 1 is immersed in the saline and incubated 2 to 4 hours at 37 C.
  • the identification of the serological group to which the organism belongs is readily determined by reference to the aforementioned table.
  • a diagnostic preparation for the identification of the serological groups of fi-hemolytic streptococci consisting essentially of a carrier having:
  • a second area comprising a hydrophobic carrier separating the first area from the third area.
  • (C) A third area impregnated with salicin and a pH indicator system capable of indicating pH changes from about 9 to 5.
  • (E) A fifth area impregnated with esculin and a member selected from the group consisting of a ferric salt, an aluminum salt and a lead salt.
  • a diagnostic preparation according to claim 1 wherein said indicator system in the glycerol area is a member selected from the group consisting of chlorophenol red, bromocresol purple, bromophenol red, bromothymol blue, neutral red, a-naphtholphthalein and a phenol red.
  • said indicator system in the salicin area is a member selected from the group consisting of chlorophenol red, bromocresol purple, bromophenol red, bromothymol blue, neutral red, ot-naphtholphthalein and phenol red.
  • ferric salt is a member selected from the group consisting of ferric chloride, ferric ammonium citrate, ferric nitrate and ferric citrate.
  • a diagnostic preparation according to claim 1 wherein said aluminum salt is aluminum ammonium citrate.
  • a diagnostic preparation according to claim 1 wherein said lead salt is a member selected from a group consisting of lead acetate and lead nitrate.
  • a process for the serological identification of fl-hemolytic streptococci by inoculating organisms onto the glycerol area, onto the salicin area and onto the esculin area of the diagnostic preparation of claim 1; incubating the resulting system at 37 C. for about 2 to 4 hours and observing these areas for color change.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
US54277A 1970-07-13 1970-07-13 Diagnostic preparation for the identification of beta-hemolytic streptococci Expired - Lifetime US3699003A (en)

Applications Claiming Priority (1)

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US5427770A 1970-07-13 1970-07-13

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US3699003A true US3699003A (en) 1972-10-17

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US (1) US3699003A (de)
JP (1) JPS524604B1 (de)
AU (1) AU2831871A (de)
CA (1) CA937140A (de)
CH (1) CH557166A (de)
DE (1) DE2129384A1 (de)
DK (1) DK125484B (de)
FR (1) FR2097950A5 (de)
GB (1) GB1290642A (de)
IT (1) IT943592B (de)
SE (1) SE364986B (de)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3877877A (en) * 1973-10-31 1975-04-15 Us Commerce Plastic cell for mixing two liquids or a liquid and a solid
US3895914A (en) * 1972-02-04 1975-07-22 Orion Yhtymae Oy Means for isolation and detection of barbituric acid derivatives and glutethimide in biological fluids
US3964871A (en) * 1974-12-18 1976-06-22 Becton, Dickinson And Company Method and device for detecting glucose
US4042329A (en) * 1974-12-18 1977-08-16 Becton, Dickinson And Company Method and device for detecting cholesterol
US4059407A (en) * 1976-04-14 1977-11-22 Becton, Dickinson And Company Disposable chemical indicators
US4066403A (en) * 1975-06-20 1978-01-03 Eastman Kodak Company Multilayer analytical element
US4189304A (en) * 1978-10-27 1980-02-19 Miles Laboratories, Inc. Device and method for detecting myoglobin
USRE30267E (en) * 1975-06-20 1980-05-06 Eastman Kodak Company Multilayer analytical element
US4259442A (en) * 1977-10-04 1981-03-31 Laboratoire De Recherche Api S.A.R.L. Process of rapid identification of bacteria of the genus Streptococcus
US4532206A (en) * 1982-07-19 1985-07-30 Vitek Systems, Inc. β-Streptococcus selective medium
WO1987001393A1 (en) * 1985-09-09 1987-03-12 Allegheny-Singer Research Institute Dry form micronitrous acid streptococci extraction-agglutination test
US5268146A (en) * 1992-03-25 1993-12-07 Litmus Concepts, Inc. Fast response test panel
US20070105168A1 (en) * 2005-10-05 2007-05-10 Pirooz Eghtesady Diagnosis and prevention of fetal heart disease
US8313938B1 (en) 2004-04-26 2012-11-20 Hardy Diagnostics Culture medium for cultivation of microorganisms
US8778657B1 (en) 2004-04-26 2014-07-15 Hardy Diagnostics Culture medium for cultivation of microorganisms

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3597321A (en) * 1969-04-17 1971-08-03 Warner Lambert Pharmaceutical Diagnostic compostion for the differentiation of staphylococci

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU420738B1 (en) * 1966-08-15 1972-01-24 Determination of bacteria

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3895914A (en) * 1972-02-04 1975-07-22 Orion Yhtymae Oy Means for isolation and detection of barbituric acid derivatives and glutethimide in biological fluids
US3877877A (en) * 1973-10-31 1975-04-15 Us Commerce Plastic cell for mixing two liquids or a liquid and a solid
US3964871A (en) * 1974-12-18 1976-06-22 Becton, Dickinson And Company Method and device for detecting glucose
US4042329A (en) * 1974-12-18 1977-08-16 Becton, Dickinson And Company Method and device for detecting cholesterol
USRE30267E (en) * 1975-06-20 1980-05-06 Eastman Kodak Company Multilayer analytical element
US4066403A (en) * 1975-06-20 1978-01-03 Eastman Kodak Company Multilayer analytical element
US4059407A (en) * 1976-04-14 1977-11-22 Becton, Dickinson And Company Disposable chemical indicators
US4259442A (en) * 1977-10-04 1981-03-31 Laboratoire De Recherche Api S.A.R.L. Process of rapid identification of bacteria of the genus Streptococcus
US4189304A (en) * 1978-10-27 1980-02-19 Miles Laboratories, Inc. Device and method for detecting myoglobin
US4532206A (en) * 1982-07-19 1985-07-30 Vitek Systems, Inc. β-Streptococcus selective medium
WO1987001393A1 (en) * 1985-09-09 1987-03-12 Allegheny-Singer Research Institute Dry form micronitrous acid streptococci extraction-agglutination test
US4673639A (en) * 1985-09-09 1987-06-16 Allegheny-Singer Research Institute Dry form micronitrous acid streptococci extraction-agglutination test
US5268146A (en) * 1992-03-25 1993-12-07 Litmus Concepts, Inc. Fast response test panel
US8313938B1 (en) 2004-04-26 2012-11-20 Hardy Diagnostics Culture medium for cultivation of microorganisms
US8518688B1 (en) 2004-04-26 2013-08-27 Hardy Diagnostics Culture medium for cultivation of microorganisms
US8778657B1 (en) 2004-04-26 2014-07-15 Hardy Diagnostics Culture medium for cultivation of microorganisms
US20070105168A1 (en) * 2005-10-05 2007-05-10 Pirooz Eghtesady Diagnosis and prevention of fetal heart disease
US7709210B2 (en) * 2005-10-05 2010-05-04 Children's Hospital Medical Center Diagnosis and prevention of fetal heart disease
US20100172897A1 (en) * 2005-10-05 2010-07-08 Children's Hospital Medical Center Diagnosis and Prevention of Fetal Heart Disease

Also Published As

Publication number Publication date
IT943592B (it) 1973-04-10
AU2831871A (en) 1972-11-02
CH557166A (de) 1974-12-31
GB1290642A (de) 1972-09-27
DK125484B (da) 1973-02-26
JPS524604B1 (de) 1977-02-05
FR2097950A5 (de) 1972-03-03
SE364986B (de) 1974-03-11
DE2129384A1 (de) 1972-01-20
CA937140A (en) 1973-11-20

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