Neisseria bacteria species identification and beta lactamase testing methods

• US 4,390,622 A
• Filed: 09/21/1981
• Issued: 06/28/1983
• Est. Priority Date: 09/21/1981
• Status: Expired due to Fees

• Current Assignee: Garry W. Cartwright
• Original Assignee: Garry W. Cartwright
• Inventors: Cartwright, Garry W.
• Primary Examiner(s): Warden, Robert J.
• Application Number:
  US06/303,725
• Time in Patent Office: 645 Days
• Field of Search: 435/18, 435/23, 435/24, 435/34, 435/37, 435/871
• US Class Current: 435/18
• CPC Class Codes: C12Q 1/12 Nitrate to nitrite reducing bacteria
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: G01N 2333/22 from Neisseriaceae (F), e.g. Acinetobacter
• CPC Class Codes: G01N 2333/986 acting on amide bonds in cyclic amides (3.5.2), e.g. beta-lactamase (penicillinase, 3.5.2.6), creatinine amidohydrolase (creatininase, EC 3.5.2.10), N-methylhydantoinase (3.5.2.6)
• CPC Class Codes: G01N 2415/00 Assays, e.g. immunoassays or enzyme assays, involving penicillins or cephalosporins
• CPC Class Codes: Y10S 435/871 Neisseria

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.

Claims

1. A method for providing rapid, high intensity, visual indication of the presence of Neisseria bacteria species in a specimen comprising:

the steps of admixing the specimen with a peptone buffer and pH indicator reagent system to form a specimen suspension, contacting the reagent system specimen suspension with at least one carbohydrate substrate taken from the class consisting of glucose, maltose, sucrose and lactose, permitting the reagent system specimen to react with the contacted carbohydrate substrate, and visually observing the reaction results for color change in a predetermined time period.

2. The method of claim 1 wherein a portion of the reagent system specimen suspension is contacted with each of said carbohydrate substrates for reaction of the reagent system specimen with the contacted carbohydrate substrate for visual observation of the reaction results for color change.

3. The method of claim 1 wherein the reaction results are visually observed within a time period of from between about two hours and about seven hours for color change.

4. The method of claim 3 wherein a color change to a yellow color indicates the presence of Neisseria bacteria species in the specimen.

5. The method of claim 4 wherein the Neisseria bacteria species undergoing testing is taken from the class consisting of N. gonorrhoeae, N. meningitidis, N. lactamica, N. sicca and N. catarrhalis.

6. The method of claim 1 wherein the peptone buffer and pH indicator reagent system has a pH within the range between about 7.1 and about 7.5.

7. The method of claim 6 wherein the peptone buffer is a proteose peptone material.

8. The method of claim 7 wherein the proteose peptone buffer is a beef peptone material.

9. The method of claim 8 wherein the proteose beef peptone buffer is peptone number 3.

10. The method of claim 7 wherein the proteose peptone is present in the reagent system in an amount of from between about 1.5% to about 2.5% by volume.

11. The method of claim 7 wherein the pH indicator is phenol red.

12. The method of claim 10 wherein the pH indicator is phenol red and is present in said aqueous reagent system in an amount of from between about 0.04 and about 0.085 grams per liter.

13. The method of rapid, high intensity, visual indication of the presence of beta lactamase enzymes in a specimen comprising the steps of admixing the specimen with a peptone buffer and pH indicator reagent system to form a specimen suspension, contacting the reagent system specimen suspension with a penicillin substrate, permitting the reagent system specimen to react with the contact penicillin substrate, and visually observing the reaction results for color change in a predetermined time period.

14. The method of claim 13 wherein the penicillin substrate is a salt of penicillin and the reaction results are visually observed within a time period of from between about five minutes and about seven hours for color change.

15. The method of claim 14 wherein the penicillin salt is taken from the class consisting of sodium and potassium penicillin G and wherein a color change to a yellow color indicates the presence of beta lactamase enzyme in the specimen.

16. The method of claim 15 wherein the Neisseria bacteria species undergoing testing is taken from the class consisting of N. gonorrhoeae, N. meningitidis, N. lactamica, N. sicca and N. catarrhalis.

17. The method of claim 13 wherein the peptone buffer and pH indicator reagent system has a pH within the range between about 7.1 and about 7.5.

18. The method of claim 17 wherein the peptone buffer is a proteose peptone material.

19. The method of claim 18 wherein the proteose peptone buffer is a beef peptone material.

20. The method of claim 19 wherein the proteose beef peptone buffer is peptone number 3.

21. The method of claim 18 wherein the proteose peptone is present in the reagent system in an amount of from between about 1.5% to about 2.5% by volume.

22. The method of claim 18 wherein the pH indicator is phenol red.

23. The method of claim 21 wherein the pH indicator is phenol red and is present in said aqueous reagent system in an amount of from between about 0.04 and about 0.085 grams per liter.

24. A method of rapid, high intensity, visual indication of the presence of Neisseria bacteria species and of beta lactamase enzyme in a specimen comprising the steps of admixing the specimen with a peptone buffer and pH indicator reagent system to form a specimen suspension, which is contacted with a penicillin substrate, and with carbohydrate substrates separately, permitting the reagent system specimen to react with the contacted carbohydrate substrates and the penicillin substrate, and visually observing the reaction results for color change induced by the presence of one or more enzymes taken from the class consisting of N. gonorrhoeae, N. meningitidis, N. lactamica, and N. sicca, in a predetermined time period.

26. The method of claim 24 wherein the reaction results are visually observed within a time perod of from between about two hours and about seven hours for color change.

27. The method of claim 26 wherein color change to a yellow color indicates the presence of Neisseria bacteria species and beta lactamase in the specimen.

28. The method of claim 27 wherein the Neisseria bacteria species undergoing testing is taken from the class consisting of N. gonorrhoeae, N. meningitidis, N. lactamica, N. sicca and N. catarrhalis.

29. The method of claim 24 wherein the peptone buffer and pH indicator reagent system has a pH within the range between about 7.1 and about 7.5.

30. The method of claim 29 wherein the peptone buffer is a proteose peptone material.

31. The method of claim 30 wherein the proteose peptone buffer is a beef peptone material.

32. The method of claim 31 wherein the proteose beef peptone buffer is peptone number 3.

33. The method of claim 30 wherein the proteose peptone is present in the reagent system in an amount of from between about 1.5% to about 2.5% by volume.

34. The method of claim 30 wherein the pH indicator is phenol red.

35. The method of claim 33 wherein the pH indicator is phenol red and is present in said aqueous reagent system in an amount of from between about 0.04 and about 0.085 grams per liter.

25. The method of claim 25 wherein a portion of the reagent system specimen suspension is contacted with each of said carbohydrate substrates for reaction of the reagent system specimen with the contacted carbohydrate substrate for visual observation of the reaction results for color change.