502 APPENDIX 



ANTIFORMIN METHOD 



For the detection of B. Tuberculosis. 



Antiformin was introduced into bacteriological 

 technique by Uhlenhuth in 1908 for the purpose of 

 demonstrating tubercle bacilli when present in small 

 numbers, in sputum or other material. It is a 

 powerful oxidising agent and rapidly destroys most 

 bacteria, but tubercle and other acid-fast organisms 

 resist its lethal action for considerable periods, and 

 upon this fact the method is based. 



To prepare Antiformin measure out and mix : 



Eau de Javelle (Liquor sodas chlorinatae B.P.) 50 c.c. 

 Sodic hydrate 15 per cent, aqueous solution .... 50 c.c. 



METHOD. 



1. Introduce the sputum or other material (e.g. 

 milk deposit and cream; pus; minced gland or other 

 organ ; caseous material ; broken down foci, etc.) into 

 a sterile tube and then add an equal volume of anti- 

 formin. 



2. Close the tube with a rubber cork and shake 

 vigorously (a sample of antiformin that does not 

 "foam" at this stage is of little use). Disintegration 

 of the material at once starts, associated bacteria are 

 destroyed and the mixture rapidly becomes a homo- 

 genous but turbid fluid a process which may be 

 hastened by : 



3. Placing the tube in the incubator at 37 C. for 30 

 minutes shaking from time to time. 



4. Centrifugalise the fluid thoroughly, at high 

 speed. 



5. Pipette off the supernatant fluid, fill up with 

 sterile distilled water, cork the tube and shake to dis- 

 tribute the deposit throughout the water. Again 

 centrifugalise. 



6. Repeat steps 4 and 5 twice more. 



