Overview of Biochemical tests used to identify bacteria in Microbiology laboratory

Overview of Biochemical tests for differentiating Gram positive cocci
Overview of Biochemical tests for differentiating Gram positive cocci

This post was most recently updated on August 24th, 2019

In this blog post, i am sharing information about common, conventional biochemical tests and their primary uses in Microbiology laboratory for the identification of particular pathogen of interest. Some of the biochemical tests mentioned here for example catalase test, oxidase test assists us in many ways apart from the primary purpose mentioned in this post;

  1. Beta-glucuronidase test (MUG Test) : To identify Escherichia coli. Escherichia coli produces the enzyme β-D-glucuronidase, which hydrolyzes β-D-glucopyranosid-uronic derivatives to aglycons and D-glucuronic acid.

    Overview of Biochemical tests for differentiating Gram positive cocci
    Overview of Biochemical tests for differentiating Gram positive cocci
  2. Bacitracin Sensitivity Test: Bacitracin sensitivity test differentiates Streptococcus pyogenes (positive) from other beta hemolytic streptococci (resistant).
  3. Bile solubility test : To differentiate Streptococcus pneumoniae from other alpha hemolytic streptococci. Bile or a solution of a bile salt, such as sodium desoxycholate rapidly lyses pneumococcal colonies.

    CAMP Test
    CAMP test :
    A. Streptococcus agalactiae (positive)
    B. Streptococcus pyogenes (Negative)
    Image Source: ASM
  4. CAMP Test: Certain organisms such as Streptococcus agalactiae (Group B streptococci), produce a diffusible extracellular protein (CAMP) factor that acts synergistically with the beta-lysin of Staphylococcus aureus and  causes enhanced lysis of RBCs.
  5. Catalase test: To differentiate Staphylococci (catalase positive) from Streptococci (catalase test negative)
  6. Citrate utilization test: To differentiate members of Enterobacteriaceae family.
  7. Coagulase test: Coagualse test is used to identify Staphylococcus aureus. Coagulase test differentiates Staphylococcus aureus (positive) from coagulase negative staphylococci (CONS), such as S. epidermidis, S. saprophyticus.
  8. DNase test: This test is used to determine the ability of an organism to hydrolyze DNA. It is primarlly used to identify Staphylococcus aureus
  9. Indole test: This test is used to determine the ability of an organism to split tryptophan to form the compound indole. It is used differentiate gram negative rods particularly E. coli in microbiology laboratory.
  10. Litmus milk decolorization test: To help identify Enterococcus and some Clostridia which have ability to metabolize litmus milk.
  11. Lysine Decarboxylase test: To assist in the identification of Salmonella and Shigella
  12. Oxidase test: To help identify Neisseria, Pasteurella, Vibrio, and Pseudomonas. This test is used to determine the presence of bacterial cytochrome oxidase.
  13. Urease test: Urease test is used to determine the ability of an organism to produce urease (an enzyme) which hydrolyzes urea. This test is done to help identify Proteus, Morganella, Yersinia enterocolitica, and Helicobacter pylori
About tankeshwar 387 Articles
Hello, thank you for visiting my blog. I am Tankeshwar Acharya. Blogging is my passion, I am working as an Asst. Professor and Microbiologist at Department of Microbiology and Immunology, Patan Academy of Health Sciences, Nepal. If you want me to write about any posts that you found confusing/difficult, please mention in the comments below.


    • Pranab Roy
      Thank you for your question. Why you need to differentiate member of Enterobacteriaceae family from that of Rhizobia? I think you never encounter such problem while identifying clinical isolate. Rhizobia are non-fermentative and forms dry, tenacious colonies.

  1. sir i am persuing msc. biochemistry and its my 4th semester . . . can u please suggest me topics for dissertation?

  2. this site is amazing. thank you so much. this help me a lot in my preparation for ASCP exam. more power and God bless

  3. It’s quite clear to understand the basic idea of biochemical tests and expected results, but I wanted to know the limitations in these tests, specifically for oxidase, catalase. If it possible plz give a link even possible

  4. Thanks for the concise and beautiful write up. Please i’ll like to know the biochemical tests for lipase producing microorganisms. Thank you.

  5. please give me detail about the ATCC CULTURE (MICROBIOLOGICS)
    what does it means and how to find out following( if it is on certificate of analysis)
    purity : 0.1% total pellet cfu
    Recovery : >1000 cfu / pellet bacillus subtilis

Do you have any queries? Please leave me in the comments section below. I will be happy to read your comments and reply.

This site uses Akismet to reduce spam. Learn how your comment data is processed.