Disposable inoculation device and process of using same

• US 4,077,845 A
• Filed: 04/20/1977
• Issued: 03/07/1978
• Est. Priority Date: 04/20/1977
• Status: Expired due to Term

• Current Assignee: Miles Laboratories, Inc. (Bayer AG)
• Original Assignee: Miles Laboratories, Inc. (Bayer AG)
• Inventors: Johnson, Leighton Clifford
• Primary Examiner(s): Tanenholtz, Alvin E.
• Application Number:
  US05/789,203
• Time in Patent Office: 321 Days
• Field of Search: 195/103.5 K, 195/127, 195/103.5 M
• US Class Current: 435/33
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/40 Manifolds; Distribution pieces fluid transfer means B01L3/563
• CPC Class Codes: C12M 33/04 by injection or suction, e.g. using pipettes, syringes, needles pipettes in general B01L3/02
• CPC Class Codes: C12M 33/06 for multiple inoculation or multiple collection of samples
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/18 Testing for antimicrobial activity of a material
• CPC Class Codes: G01N 2035/00148 Test cards, e.g. Biomerieux or McDonnel multiwell test cards

Abstract

A self-contained rehydratable microtiter type device is disclosed in which concentrations of suitable agents are predeposited and dried in growth wells and are then rehydrated with aliquot amounts of the inoculum to be tested. The apparatus and system permit the detection, identification and grouping of urinary tract pathogens.

Claims

1. Inoculation apparatus comprising a plate formed with at least one sealed growth well, said plate being formed with a sealed reservoir, at least one siphon tube projecting into said reservoir and terminating adjacent the bottom thereof, flow passage means communicating between said siphon tube and an upper portion of said growth well, said plate being provided with aperture means communicating with said reservoir and having a first portion through which liquid inoculum can be introduced into said reservoir, said aperture means also having a second portion adapted for connection with pump means, and removable closure means for sealingly closing each of said first and second portions of said aperture means following connection of said second aperture means portion with pump means and after introduction into said reservoir of an amount of liquid inoculum sufficient to provide a liquid level in said reservoir above the lower end of said siphon tube, withdrawal by said pump means of a predetermined volume of air from said reservoir causing evacuation of air from said growth well, flow passage means and siphon tube, and the subsequent introduction of a predetermined quantity of air into said reservoir by said pump means causing liquid inoculum to be forced from said reservoir through said siphon tube and passage means to said growth well.

2. The inoculation apparatus of claim 1 in which the first portion of said aperture means comprises a filling port formed in the upper surface of said plate.

3. The inoculation apparatus of claim 1 in which the second portion of said aperture means comprises an aspiration port formed in the upper surface of said plate.

4. The inoculation apparatus of claim 1 in which the passage means comprise grooves formed in the upper surface of said plate which are covered by a layer of pressure sensitive tape means sealed to the upper surface of said plate.

5. The inoculation apparatus of claim 1 in which a plurality of growth wells are present connected by equidistant flow passage means to said siphon tube.

6. The inoculation apparatus of claim 1 in which at least a wall portion of said growth well is transparent.

7. The inoculation apparatus of claim 1 having a plurality of growth wells comprising recesses formed in the upper surface of said plate, said recesses being covered by transparent layer of pressure sensitive tape means sealed to the upper surface of said plate.

8. The inoculation apparatus of claim 1 in which said reservoir comprises a recess formed in the bottom surface of said plate, said recess being covered by a layer of pressure sensitive tape sealed to the bottom surface of said plate.

9. Inoculation apparatus comprising:

a plate having upper and lower surfaces;

a central manifold chamber disposed along the lower surface of said plate;

a filling port located in the upper surface of said plate, said filling port being interconnected with said central manifold chamber;

siphon tubes extending from near the upper surface of said plate into said central manifold chamber;

separate growth wells located in the upper surface of said plate;

a plurality of grooves in the upper surface of said plate, each of said grooves having an outlet end connected with a growth well and an inlet end interconnected with a siphon tube;

an aspiration bore located in said upper surface, said aspiration bore being connected with said central manifold chamber and adapted to being connected to means for applying a vacuum to said growth wells;

first covering means having a first and second tab at one end, said first covering means sealingly engaging said upper surface of said plate thereby forming with said grooves a plurality of sealed tube-like channels connecting the siphon tubes with the growth wells and forming a cover for said growth wells, filling port and aspiration bore, said first covering means being split at one end so as to enable the first covering means to be pulled back by means of said first tab from said filling port or by means of said second tab from the aspiration bore and then being resealed;

a second covering means sealingly engaging said lower surface of the plate thereby forming a cover on said central manifold chamber.

10. The apparatus of claim 9 wherein said first and second transparent covering means comprise separate sheets of clear plastic adhesively mounted to the upper and lower surfaces, respectively.

11. The apparatus according to claim 10 wherein said sheets of clear plastic are made from transparent pressure sensitive tape.

12. Apparatus for conducting antibiotic susceptibility tests on clinical specimens, said apparatus comprising:

an elongated plate having substantially parallel upper and lower surfaces,

a central manifold chamber disposed along a portion of the lower surface of said plate;

a filling port located in the upper surface of said plate, which port is connected with said central manifold chamber;

an aspiration bore located adjacent to said filling port in the upper surface of said plate, said aspiration bore being connected with said central manifold chamber;

siphon tubes extending perpendicularly from near the upper surface of said plate into said central manifold chamber;

individual detection wells disposed along the upper surface of said plate;

filler passageways leading from said siphon tubes to said detection wells;

means for isolating said detection wells, said filler passageways, said filling port and said aspiration bore from the surrounding atmosphere on the upper surface of said plate;

means for isolating the central manifold chamber from the surrounding atmosphere on the lower surface of said plate; and

a blend of culture medium and known antibiotics in at least some of the detection wells, the culture medium being sensitive to a microorganism such that the optical characteristics of the culture medium changes when the microorganism is sustained in the culture medium, said culture medium further being selective in that it undergoes a change in optical characteristics only when a specific microorganism is sustained by it.

13. Apparatus for conducting antibiotic susceptibility tests on clinical specimens, which apparatus comprises:

a plate having a plurality of detection wells opening out of an upper surface thereof with the wells being visible on both sides of the plate so light can pass through the apparatus at the location of the wells, said plate also having a filling port, an aspiration bore and a central manifold chamber, said filling port and aspiration bore being interconnected with said central manifold chamber such that clinical specimens can be poured into the central manifold chamber through the filling port and air can be evacuated from the detection wells through the aspiration bore;

siphoning means extending from the upper surface in said plate down into said central manifold chamber and filler passageways leading from said siphoning means to the detection wells, said siphoning means and said filler passageways adapted to transmit clinical specimens from said central manifold chamber to said detection wells;

a selective culture medium in said detection wells with known antibiotics blended with the culture medium in at least some of the wells, the culture medium being selective in the sense that the light transmitting characteristics of the culture medium will change when the clinical specimen contains a microorganism to which the culture medium is specific, and

transparent closing means applied to surfaces of said plate for isolating the plate from the surrounding atmosphere.

14. A process for determining the susceptibility of specific microorganisms to antibiotics, said process comprising:

removing a first tape member from one end of a microtiter tray formed with a sealed manifold chamber, separate growth wells, at least one siphon tube projecting into said manifold chamber and terminating adjacent the bottom thereof, flow passage means communicating between said siphon tube and upper portion of said growth wells, and aperture means communicating with said manifold chamber, said aperture means having a first filling portion covered by said first tape member and a second portion covered by a second tape member, said second portion adapted for connection with pump means;

introducing a clinical specimen containing microorganisms into said central manifold chamber of said tray;

resealing the first tape member and removing a second tape member from said microtiter tray;

evacuating air from said growth wells in which a selective culture medium is contained and from wells in which blends of the selective culture medium and known antibiotics are contained, said culture medium favoring the specific microorganism such that the light transmitting characteristics of a mixture of the culture medium and water will change when the favored microorganism is sustained within and nourished by the medium;

replacing the evacuated air with an aliquot mixture of said clinical specimen from said central manifold chamber and resealing said second tape member;

incubating the tray; and

observing the growth wells for change in the light transmitting characteristics thereof.

15. The process according to claim 14 wherein the culture medium is freeze-dried and water present in the clinical specimen rehydrates the culture medium.

16. The process according to claim 14 wherein change in the light transmitting characteristics of the growth wells is observed by projecting light through said wells and measuring the intensity of the light passing through said growth wells.