Claims for Patent: 4,582,807

Summary for Patent: 4,582,807

Title:	Cultivation medium for mycobacteria and use thereof
Abstract:	A medium for the rapid cultivation of mycobacteria, in particular leprosy and tubercle bacilli, comprises amino acids, sugars, phospholipids, muscle metabolism compounds, vitamins, inorganic salts and trace minerals.
Inventor(s):	Veeraraghavan; Natteri (Madras 600090, IN)
Assignee:
Application Number:	06/403,827
Patent Claims:	1. A medium suitable for the cultivation of mycobacteria consisting essentially of (a) from 2 to 2.5 grams per liter of an acidic amino acid selected from the group consisting of aspartic acid, asparagine, glutamic acid and mixtures thereof (b) a total of 3.4 to 4.1 grams per liter of neutral amino acid selected from the group consisting of .beta.-alanine, DL-.alpha. alanine, phenylalanine, tyrosine, cystine, cysteine, proline, serine, leucine methionine and mixtures thereof (c) 120-160 mg per liter of arginine (d) a total of 350-420 mg per liter of "muscle metabolism compound" selected from the group consisting of glycogen, glutathione, adenylic acid, creatine, sarcosine, carnatine, carnosine, glycocyamine, adenosine diphosphate, adenosine triphosphate, phosphocreatin and mixtures thereof (e) a total of from 1 to 1.5 grams/liter of monosaccharaide sugar selected from the group consisting of glucosamine, mannose, arabinose, galactose, D-ribose and mixtures thereof (f) from 40-50 mg/liter of adenosine, cytosine or a mixture of the two (g) a total of from 5 to 10 mg/liter of phospholipid selected from the group consisting of lecithin, phosphotidyl ethanolamine, phosphotidyl inositol, and mixtures thereof (h) 5-15 mg/liter cholesterol (i) 0.4-0.6 mg/liter linoleic acid (j) 1-1.5 mg/liter oleic acid (k) 4-6 mg/liter histamine (l) 4-6 mg/liter heparin (m) a total of 3.5-5 mg/liter of a vitamin selected from the group consisting of vitamins B, D, E, K and mixtures thereof (n) 4-6 g/liter dextran (o) 2-3 mg/liter protamine sulfate (p) 6-10 g/liter inorganic salts and (q) trace minerals in sterile water. 2. A medium according to claim 1, wherein the pH is in the range 7.2-7.4. 3. A medium according to claim 1, wherein the acidic amino acids are present in a ratio 4-6:4-6:1 of L-aspartic acid: asparagine: L-glutamic acid. 4. A medium according to claim 1, wherein the amounts of neutral amino acids present are: .beta.-alanine: 1.35-1.85 g/liter DL-.alpha. alanine: 0.4-0.6 g/liter L-proline: 0.08-0.12 g/liter DL-serine: 0.08-0.12 g/liter L-leucine: 0.08-0.12 g/liter phenyl alanine: 0.06-0.08 g/liter methionine: 0.025-0.035 g/liter cystine: 0.45-0.5 g/liter tyrosine: 0.65-0.8 g/liter cysteine: 0.15-0.25 g/liter. 5. A medium according to any one of claims 1, 2, 3 or 4, which further contains an amino acid selected from the group consisting of L-glycine, .gamma.-aminobutyric acid, taurine, glycylgylcine carnatine and mixtures thereof. 6. A medium according to claims 1, 2, 3 or 4, wherein 95-100% by weight of the said monosaccharide sugar component consists of glucosamine, galactose and arabinose each one of said sugars being present in a substantially equal amount by weight. 7. A medium according to claims 1, 2, 3 or 4, wherein 75-100% of the "muscle metabolism compounds" is a mixture of glycogen and glutathione in a substantially 2:1 weight ratio. 8. A method for cultivating leprosy bacilli which comprises introducing said bacilli into a medium consisting essentially of (a) from 2 to 2.5 grams per liter of an acidic amino acid selected from the group consisting of aspartic acid, asparagine, glutamic acid and mixtures thereof (b) a total of 3.5 to 4.1 grams per liter of neutral amino acid selected from the group consisting of .beta.-alanine, DL-.alpha. alanine, pheylalanine, tyrosine, cystine, cysteine, proline, serine, leucine methionine and mixtures thereof (c) 120-160 mg/per liter of arginine (d) a total of from 350-420 mg per liter of "muscle metabolism compound" selected from the group consisting of glycogen, glutathione, adenylic acid, creatine, sarcosine, carnatine, carnosine, glycocyamine, adenosine diphosphate, adenosine triphosphate, phosphocreatin and mixtures thereof (e) a total of from 1 to 1.5 grams liter of monosaccharide sugar selected from the group consisting of glucosamine, mannose, arabinose, galactose, D-ribose and mixtures thereof (f) from 40-50 mg/liter of adenosine, cytosine or a mixture of the two (g) a total of 5 to 10 mg/liter of phospholipid selected from the group consisting of lecithin, phosphotidyl ethanolamine, phosphotidyl inositol, and mixtures thereof (h) 5-15 mg/liter cholesterol (i) 0.4-0.6 mg/liter linoleic acid (j) 1-1.5 mg/liter oleic acid (k) 4-6 mg/liter histamine (l) 4-6 mg/liter heparin (m) a total of 3.5-5 mg/liter of a vitamin selected from the group consisting of vitamins B, D, E, K and mixtures thereof (n) 4-6 g/liter dextran (o) 2-3 mg/liter protamine sulfate (p) 6-10 g/liter inorganic salts and (q) trace minerals in sterile water and maintaining said medium at a temperature of 0.degree.-10.degree. C. 9. A method for diagnosis leprosy which comprises placing skin scrapings from suspected leper in a medium consisting essentially of (a) from 2 to 2.5 grams per liter of an acidic amino acid selected from the group consisting of aspartic acid, asparagine glutamic acid and mixtures thereof (b) a total of from 3.5 to 4.1 grams per liter of neutral amino acid selected from the group consisting of .beta.-alanine, DL-.alpha. alanine, phenylanine, tyrosine, cystine, cysteine, proline, serine, leucine methionine and mixtures thereof (c) 120-160 mg per liter of arginine (d) a total of from 350-420 mg per liter of "muscle metabolism compound" selected from the group consisting of glycogen, glutathione, adenylic acid, creatine, sarcosine, carnatine, carnosine, glycocyamine, adenosine diphosphate, adenosine triphosphate, phosphocreatin and mixtures thereof (e) a total of from 1 to 1.5 grams/liter of monosaccharide sugar selected from the group consisting of glucosamine, mannose, arabinose, galactose, D-ribose and mixtures thereof (f) from 40-50 mg/liter of adenosine, cytosine or a mixture of the two (g) a total of from 5 to 10 mg/liter of phospholipid selected from the group consisting of lecithin, phosphotidyl ethanolamine, phosphotidyl inositol and mixtures thereof (h) 5-15 mg/liter cholesterol (i) 0.4-0.6 mg/liter linoleic acid (j) 1-1.5 mg/liter oleic acid (k) 4-6 mg/liter histamine (l) 4-6 mg/liter heparin (m) a total of 3.5-5 mg/liter of a vitamin selected from the group consisting of vitamins B, D, E, K and mixtures thereof (n) 4-6 g/liter dextran (o) 2-3 mg/liter protamine sulfate (p) 6-10 g/liter inorganic salts and (q) trace minerals in sterile water and thereafter cultivating the scrpaings in said medium at 0.degree.-10.degree. C. and periodically determining by staining followed by visual observation whether the number of single bacilli present is increasing. 10. A method for testing the sensitivity of leprosy bacillus to a particular drug which comprises cultivating leprosy bacillus in a medium consisting essentially of (a) from 2 to 2.5 grams per liter of an acidic amino acid selected from the group consisting of aspartic acid, asparagine, glutamic acid and mixtures thereof (b) a total of from 3.5 to 4.1 grams per liter of neutral amino acid selected from the group consisting of .beta.-alanine, DL-.alpha. alanine, phenylalanine, tyrosine, cystine, cysteine, proline, serine, leucine methionine and mixtures thereof (c) 120-160 mg per liter of arginine (d) a total of from 350-420 mg per liter of "muscle metabolism compound" selected from the group consisting of glycogen, glutathione, adenylic acid, creatine, sarcosine, carnatine, carnosine, glycocyamine, adenosine diphosphate, adenosine triphosphate, phosphocreatin and mixtures thereof (e) a total of from 1 to 1.5 grams/liter of monosaccharide sugar selected from the group consisting of glucosamine, mannose, arabinose, galactose, D-ribose and mixtures thereof (f) from 40-50 mg/liter of adenosine, cytosine or a mixture of the two (g) a total of from 5 to 10 mg/liter of phospholipid selected from the group consisting of lecithin, phospotidyl ethanolamine, phosphotidyl inositol, and mixtures thereof (h) 5-15 mg/liter cholesterol (i) 0.4-0.6 mg/liter linoleic acid (j) 1-15 mg/liter oleic acid (k) 4-6 mg/liter histamine (l) 4-6 mg/liter heparin (m) a total of 3.5-5 mg/liter vitamin selected from the group consisting of vitamins B, D, E, K and mixtures thereof (n) 4-6 g/liter dextran (o) 2-3 mg/liter portamine sulfate (p) 6-10 g/liter inorganic salts and (q) trace minerals in sterile water in the presence of the test drug and determining by staining followed by visual observation whether the number of single bacilli present is increasing. 11. A method for diagnosing tuberculosis which comprises cultivating a specimen from one suspected of being tubercular in a medium consisting essentially of (a) from 2 to 2.5 grams per liter of an acidic amino acid selected from the group consisting of aspartic acid, asparagine, glutamic acid and mixtures thereof (b) a total of from 3.5 to 4.1 grams per liter of neutral amino acid selected from the group consisting of .beta.-alanine, DL-.alpha. alanine, phenylalanine tyrosine, cystine, cysteine, proline, serine, leucine methionine and mixtures thereof (c) 120-160 mg per liter of arginine (d) a total of from 350-420 mg per liter of "muscle metabolism compound" selected from the group consisting of glycogen, glutathione, adenylic acid, creatine, sarcosine, carnatine, carnosine, glycocyamine, adenosine diphosphate, adenosine triphosphate, phosphocreatin and mixtures thereof (e) a total of from 1 to 1.5 grams/liter of monosaccharide sugar selected from the group consisting of glucosamine, mannose, arabinose, galactose, D-ribose and mixtures thereof (f) from 40-50 mg/liter of adenosine, cytosine or a mixture of the two (g) a total of from 5 to 10 mg/liter of phospholipid selected from the group consisting of lecithin, phosphotidyl ethanolamine, phosphotidyl inositol and mixtures thereof (h) 5-15 mg/liter cholesterol (i) 0.4-0.6 mg/liter linoleic acid (j) 1-1.5 mg/liter oleic acid (k) 4-6 mg/liter histamine (l) 4-6 mg/liter heparin (m) a total of 3.5-5 mg/liter of a vitamin selected from the group consisting of vitamins B, D, E, K and mixtures thereof (n) 406 g/liter dextran (o) 2-3 mg/liter protamine sulfate (p) 6-10 g/liter inorganic salts and (q) trace minerals in sterile water at about 37.degree. C. and periodically determining by staining followed by visual observation whether the number of tubercule bacilli present has increased. 12. A method for testing the sensitivty of tubercule bacilli to a particular drug which comprises cultivating tubercule bacilli in a medium consisting essentially of (a) from 2 to 2.5 grams per liter of an acidic amino acid selected from the group consisting of aspartic acid, asparagine glutamic acid and mixtures thereof (b) a total of from 3.5 to 4.1 grams per liter of neutral amino acid selected from the group consisting of .beta.-alanine, DL-.alpha. alanine, phenylalanine, tyrosine, crystine, cysteine, proline, serine, leucine methionine and mixtures thereof (c) 120-160 mg per liter of arginine (d) a total of from 350-420 mg per liter of "muscle metabolism compound" selected from the group consisting of glycogen, glutathione, adenylic acid, creatine, sarcosine, carnatine, carnosine, glycocyamine, adenosine diphosphate, adenosine triphosphate phosphocreatin and mixtures thereof (e) a total of from 1 to 1.5 grams/liter of monosaccharide sugar selected from the group consisting of glucosamine, mannose arabinose, galactose, D-ribose and mixtures thereof (f) from 40-50 mg/liter of adenosine, cytosine or a mixture of the two (g) a total of from 5 to 10 mg/liter of phospholipid selected from the group consisting of lecithin, phosphotidyl ethanolamine, phosphotidyl inositol, and mixtures thereof (h) 5-15 mg/liter cholesterol (i) 0.4-0.6 mg/liter linoleic acid (j) 1-1.5 mg/liter oleic acid (k) 4-6 mg/liter histamine (l) 4-6 mg/liter heparin (m) 3.5-5 mg/liter of a vitamin selected from the group consisting of vitamins B, D, E, K and mixtures thereof (n) 4-6 g/liter dextran (o) 2-3 mg/liter protamine sulfate (p) 6-10 g/liter inorganic salts and (q) trace minerals in sterile water in the presence of the test drug and periodically determining by staining followed by visual observation whether the number of tubercle bacilli present has increased.

Details for Patent 4,582,807

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Jubilant Hollisterstier Llc	N/A	positive skin test control-histamine	Injection	103891	03/13/1924		2039-09-23
Merck Sharp & Dohme Corp.	ZOSTAVAX	zoster vaccine live	For Injection	125123	05/25/2006		2039-09-23