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How do you do Voges-Proskauer test properly? A step by step procedure and explanation of each...

How do you do Voges-Proskauer test properly? A step by step procedure and explanation of each step would be nice. I have checked online but there seems to be a variety of way. I am asking for someone who has done it professionally or have done it many times.

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Expert Solution

Voges- Proskauer test is used to ddtect the presence of enteriobacteria. It is done by two methods - long and rapid test.

Procedure:

  • Take a tube. Add overnight grown culture on nutrient agar, MacConkey agar, Blood agar or Chocolate agar.
  • Incubate for 24 to 48 hours at 35 C
  • Take 1 ml of the incubated solution in another test tube.
  • The reagents should be in room temperature.
  • Add 0.6 mL (9 drops) of ?-Naphthol Reagent to the 1ml solution.
  • Add 0.2 mL (3 drops) of 40% Potassium Hydroxide.
  • Shake the tube gently for 30 seconds.
  • Expose the broth to open air for oxygen reaction so that there will be colour change.
  • Allow tube to stand.

Rapid test:

  • Take a test tube. Add the culture grown on Nutrient agar, MacConkey agar, Blood agar, or Chocolate agar.
  • Add 0.2 ml of broth to this.
  • Incubate at 35 C using a water bath for 4 hours.
  • Add 2 drops of creatine reagent and mix gently to the test tube.
  • Then add 3 drops of ?-Naphthol Reagent and mix gently.
  • Then add 3 drops of 40% Potassium Hydroxide and gently mix for 10 seconds.
  • Let the tube stand for 15 minutes.
  • After 15 minutes, interpret the colour change.

Interpretation:

  • Positive: Acetoin present if it is red or pink colour on top of medium.
  • Negative: Acetoin absent if it yellow or copper colour on top of medium.

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