Title of Invention

MICROBIOLOGICAL MEDIUM FOR RAPID CULTURE OF TUBERCULOSIS BACILLI

Abstract

This invention relates to an improved microbiological culture medium for growth of tuberculosis bacilli where they can produce visible growth within 10 days instead of 3-6 weeks time which is required for their visible growth on conventional media. The medium is comprised of Disodium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulphate, sodium citrate, asparagine, glycerol, sodium metasilicate, phosphoric acid, fatty acid synthase and distilled water. The addition of sodium metasilicate and fatty acid synthase in a medium for culture of tuberculosis bacilli is new.

Full Text

2 Contd... This invention relates to a microbiological culture medium for culture of tuberculosis bacilli, causative agent of tuberculosis and other related microorganisms. In the known art the culture media which are used for culture of tuberculosis bacilli takes 3-6 weeks time for the culture to grow, thus in most of the patients by the time the culture reports are available the patients either will lead to fulminating type tuberculosis or has developed many complications. Also there are imported computerized automated analyzers for culture of tuberculosis bacilli but they are too costly, difficult to maintain and thus are useless for people in India, as most of the patients cannot afford even the basic cost of the reagents for that instrument. The conventional media used for culture of tuberculosis bacilli contain different organic and inorganic salts along with various other nutrients like egg, which is very difficult to incorporate and standardize in the medium. The main difference with this new medium is that this new medium is solidified with sodium metasilicate and phosphoric acid where silicate not only helps in solidification but it also enhances growth of tuberculosis bacilli. Sodium metasilicate and phosphoric acid were not used in any known conventional medium. It is the object of the invention to devise a new medium for obviating the above said prior art defects like addition of egg in the medium and to give a culture report within 10 days at a very low cost within the reach of the people in India and in other developing countries. It is also the object of the invention to have a medium which is very simple in preparation and easy for cultivation and also available at affordable price. 3 Accordingly the invention provides a new bacteriological culture medium which is to be prepared in the following ways:- I. Preparation of carbon-source solution: It consists of the following chemicals: Disodium hydrogen phosphate 0.30 Gm Potassium dihydrogen phosphate 0.40 Gm Magnesium sulphate 0.06 Gm Sodium citrate 0.25 Gm Asparagine 0.50 Gm Glycerol 2.0 ml Distilled water 50.0 ml hi a 100 ml flask, 0.30 Gm Disodium hydrogen phosphate, 0.40 Gm Potassium dihydrogen phosphate, 0.06 Gm Magnesium sulphate, 0.25 Gm Sodium citrate and 0.50 Gm Asparagine are taken after proper weighing in a chemical balance, after that 2.0 ml glycerol iis added with the help of a glass pipette and then 50 ml distilled water is added and mixed thoroughly by shaking. All chemicals are of analytical grade. Then the mixture is autoclaved at 10 lb pressure for 10 minutes. II. Preparation of sodium metasilicate solution: It consists of the following chemicals: Sodium metasilicate 15.620 Gm Distilled water 50.0 ml 4 In a chemical balance 15.62 Gm of analytical grade sodium metasilicate is weighed and taken in a 100 ml flask and then 50 ml of distilled water is mixed with it. After thorough mixing, the mixture should be autoclaved at 15 lb pressure for 15 minutes. III. Preparation of phosphoric acid solution: It consists of the following chemicals: Phosphoric acid 4.0 ml Distilled water 21.0 ml After taking 21.0 ml of distilled water in a 100 ml flask, 4.0 ml analytical grade phosphoric acid is added with it carefully with the help of a glass pipette. After thorough mixing, the mixture is autoclaved at 15 lb pressure for 15 minutes. Preparation of final medium: 2 ml of carbon source solution (I), 2 ml of sodium metasilicate solution (II), 0.8 ml of phosphoric acid solution (III) are mixed with 20 units of fatty acid synthase in a sterile capped vial with the help of sterilized pipettes. After mixing gently so that no bubble is formed in the medium, the cap of the vial is to be closed and the vial should be kept in a slanted position. The medium will be solidified within 15 minutes producing a transparent solid medium. All chemicals are of analytical grade. The pH of the final medium is 6.8. After inoculation of clinical materials e.g. sputum of patients suffering from tuberculosis of lung, the TB bacilli will grow within 10 days on this medium at 37°C in incubators. 5 I claim: 1. An improved solid medium for culture of tuberculosis bacilli as herein described. 2. An improved solid medium as claimed in claim 1 comprising of Disodium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulphate, sodium citrate, asparagine, glycerol, sodium metasilicate, phosphoric acid, fatty acid synthase and distilled water and prepared in the following manner: 2 ml of sterile liquid carbon source solution comprising of Disodium hydrogen phosphate 0.30 Gm, Potassium dihydrogen phosphate 0.40 Gm, Magnesium sulphate 0.06 Gm, Sodium citrate 0.25 Gm, Asparagine 0.50 Gm, Glycerol 2.0 ml, Distilled water 50.0 ml; 2 ml of sterile liquid sodium metasilicate solution comprising of Sodium metasilicate 15.620 Gm, Distilled water 50.0 ml; 0.8 ml of sterile liquid phosphoric acid solution comprising of Phosphoric acid 4.0 ml, Distilled water 21.0 ml and 20 units of fatty acid synthase. The final medium is solid. 3. An improved medium as claimed in claim 2 wherein the addition of sodium metasilicate and fatty acid synthase in a medium for culture of tuberculosis bacilli is new. 4. An improved medium as claimed in claim 1 wherein the tuberculosis bacilli grows on this medium within 10 days at 37°C which is less than the time required for its growth on available conventional media. This invention relates to an improved microbiological culture medium for growth of tuberculosis bacilli where they can produce visible growth within 10 days instead of 3-6 weeks time which is required for their visible growth on conventional media. The medium is comprised of Disodium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulphate, sodium citrate, asparagine, glycerol, sodium metasilicate, phosphoric acid, fatty acid synthase and distilled water. The addition of sodium metasilicate and fatty acid synthase in a medium for culture of tuberculosis bacilli is new.

Documents:

00318-cal-1999 abstract.pdf

00318-cal-1999 claims.pdf

00318-cal-1999 correspondence.pdf

00318-cal-1999 description(complete).pdf

00318-cal-1999 description(provisional).pdf

00318-cal-1999 form-1.pdf

00318-cal-1999 form-18.pdf

00318-cal-1999 form-2.pdf

00318-cal-1999 form-3.pdf

00318-cal-1999 letters patent.pdf

318-cal-1999-granted-abstract.pdf

318-cal-1999-granted-claims.pdf

318-cal-1999-granted-description (complete).pdf

318-cal-1999-granted-form 2.pdf

318-cal-1999-granted-specification.pdf