ACCRA TECHNICAL UNIVERSITY
DEPARTMENT OF MEDICAL
LABORATORY TECHNOLOGY
Course Name: MICROBIOLOGY V
Course Code: MLM 413
TOPIC: BACTERIA IDENTIFICATION TECHNIQUES
LECTURER:
DR. HENRY HACKMAN
METHODS USED IN THE IDENTIFICATION OF
BACTERIA
Colonial morphology
Microscopic morphology
Biochemical / Physiological Tests
Serological Tests
Molecular Methods (PCR, Gene Sequencing)
COLONIAL MORPHOLOGY
When viewed from above: colonies may appear round,
irregular, crenated, or branching. They may be transparent or
opaque and their surface may be smooth or rough, dull or
shiny. The colonies of capsulated species appear mucoid.
Mature colonies of pneumococci have a ringed appearance.
When viewed from the side: Colonies may appear flat or
raised in varying degrees sometimes with bevelled edges or
with a central elevation or depression.
When touched with a wire loop: some colonies are soft and
easily emulsified such as Staphylococcus aureus, whereas
others are difficult to break up such as Streptococcus
pyogenes.
COLONIAL MORPHOLOGY
COLONIAL MORPHOLOGY
The colour of colonies: this also helps to identify
bacteria, especially when using differential media
containing indicators.
Changes which may occur in the medium when
bacteria are cultured on solid agar include haemolytic
reactions, pigment production, colour changes
surrounding carbohydrate fermenting colonies and
blackening due to hydrogen sulphide production.
Haemolytic Reactions: Occurs on Blood agar. Beta
haemolytic (complete), Alpha haemolytic (partial),
Gamma haemolytic (no haemolysis)
COLONIAL MORPHOLOGY
MICROSCOPIC MORPHOLOGY
Gram Staining
Microscopy
Observe the Gram reaction, shape and arrangement
of cells.
Report: Gram positive cocci in clusters or Gram
negative rods in chains; Gram negative intracellular
diplococci (GNID)
MICROSCOPIC MORPHOLOGY
BIOCHEMICAL / PHYSIOLOGICAL TESTS
Test Principle, Test Procedure /Method, Test Type, Test Results, Test
Interpretation
Coagulase test,
Catalase test,
Bile solubility test,
Oxidase test,
Urease test,
Citrate test,
KIA/ TSI test,
IMViC test ((Indole, Methyl-Red, Vogues-Proskauer, and Citrate): GN Bacteria
COAGULASE TEST (STAPHYLOCOCCUS AUREUS AND
OTHER SPECIES)
CATALASE TEST
(STAPHYOCOCCUS AND STREPTOCOCCUS)
BILE SOLUBILITY TEST (STREPTOCOCCUS PNEUMONIAE
AND VIRIDANS)
OXIDASE TEST (PSEUDOMONAS AEROGINOSA AND
PROTEUS)
UREASE TEST (PROTEUS)
CITRATE TEST (KLEBSIELLA, CITROBACTER, SALMONELLA
SPP)
TRIPLE SUGAR IRON (TSI) AGAR OR
KLIGLER IRON AGAR (GN BACTERIA)
INDOLE TEST (E.COLI)
SEROLOGICAL TESTS
Flocculation tests,
Agglutination tests,
Complement fixation test,
Enzyme-linked Immunosorbent Assay (ELISA)
Fluorescent antibody test.
MOLECULAR METHODS
Polymerase Chain Reaction (real time DNA
amplification)
Sequencing Techniques (16S rRNA)
DNA fingerprinting
Nucleic acid hybridization methods
Whole genome sequencing
Molecular Typing of
Microorganisms(Chromatography)
