Bacterial identification procedure Part 1 : Preparation of API miniaturized system

Distilled water Physiological sterile water tube (5 mL) Oil Agar Petri dish API system with box and lid Sterile pipettes Unsteril pipette Materials avaliable

1.Bacterial suspension preparation

1.1. With a sterile inoculation loop, take a bacterial colony on agar Petri dish. WARNING! The sample is realized in asepsis around the Bunzen burner.

1.2. Make a suspension with this bacterial colony in a steril tube of physiological water (5mL). In the water, rub inoculation loop on the tube side to disperse all the bacteria. WARNING! The sample is realized in asepsis around the Bunzen burner.

1.3Homogenize your tube by manual rotations and write « S » on this tube. S

2.Preparation of the API system

2.1. Fill up the bottom of the box with distilled water to cover all the cavities Make a rotational movement with the box to distribute the water in all the cavities. After that, aspire the water excess with a unsterile pipette.

2.3. With a forceps, put the API system on the the box Cover with lid and write your «table number» on the plastic label of the box.

2.Filling the API system

Introduction Non underline test Underline test Framed test Examples : Microtube Cupule

3.1. Open the box and use the lid such as a support to place the box in an inclined position.

3.2 Homogenize your suspension by manual rotations. Take 2 mL of the suspension « S » with a sterile pipette. WARNING! The sample is realized in asepsis around the Bunzen burner. S

3.3 Fill up all the tests with this suspension respecting this next rules :  For an underline test or not (example : ONPG or ADH), fill up only the microtube,  For an framed test (example CIT), fill up the microtube and the cupule.Cupule microtube

3.4 With a sterile pipette, add oil in the 5 cupules on the underlines tests (ADH, LDC, ODC, H2S, URE). Cupule

3.5 Close the box and incubate at 37°C during 16 hours. API box in incubator

4.Analyse your results The second day, after incubation, identify your bacteria (part 2).