SOP for Identification of Organisms

1.0 Objective

To identify the organism of all cultures lying in the microbiological department or contaminants.

2.0 Scope

This procedure is applicable for morphological identification for organism of all cultures lying in microbiological laboratory and all contaminants obtained during analysis.

3.0 Responsibility

3.1 Doing: Tech. Assistant ( Microbiologist)
3.2 Checking: Executive/Manager

4.0 Accountability

Head of the Department

5.0 Procedure

5.1 Wear sterile gloves, goggles and mask before starting the work.
5.2 Take clean and dry slide and make it grease free by passing it from flame for 3-4 times.
5.3 With the help of sterile nichrome wire loop, take the growth of organisms to be identified and spread evenly on the center of slide. Allow it to air dry.
5.4 Gently pass it through flame 2-3 times for fixation of smear.
5.6 Wash with water and flood with grams iodine for one minute.
5.7 Wash the slide with water and decolorized with alcohol or acetone until no further violet colour comes off.
5.8 Wash with water.
5.9 Counter stain with 0.5 % safrannin for about half minutes.
5.10 Wash the slide with water, air dry it and observe the slide under 100 X immersion lense of microscope.
5.11 Gram + ve bacilli & gram + ve cocci are stained violet and gram -ve cocobacilli and gram -ve Cocci are stained pink in colour.
5.12 Observe the result and report it in respective format.

6.0 Abbreviations

6.1 % = Percentage
6.2 +ve = Positive
6.3 - ve = Negative
6.4 min. = Minutes

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