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Microbial Diagnostic Disks: Introduction, List of Disks, Uses, and Related Footages

Introduction of Microbial Diagnostic Disks

Microbial Diagnostic Disks make it easy to presumptive identification of organisms. They are simple and easy to handle for the identification of bacteria in the Clinical Microbiology Laboratory. Some common examples of such disks are optochin, oxidase, and bacitracin.

List of Microbial Diagnostic Disks

  • Novobiocin
  • Cefoxitin
  • Optochin
  • Oxidase disks
  • Bacitracin 10 U
  • Bacitracin 0.04 U
  • ONPG disks
  • X, V, and XV disks

Uses of Microbial Diagnostic Disks

  • Novobiocin: Staphylococcus saprophyticus is an intrinsic resistance. This disc uses to differentiate S. saprophyticus from other coagulase-negative staphylococci by the overnight incubation disc test method.
  • Optochin: It uses for the presumptive identification of Streptococcus pneumoniae, which is optochin-sensitive, from other alpha-hemolytic streptococci such as viridans streptococci, which are resistant.
  • Oxidase disks: To differentiate oxidase-positive organisms like Pseudomonas, Aeromonas, Vibrio, Haemophilus, Neisseria, Brucella, etc.
  • Bacitracin (0.04 U): It uses to differentiate Streptococcus pyogenes from other ß hemolytic streptococci, Streptococcus agalactiae . S. pyogenes is sensitive whereas S. agalcatiae is resistant.
  • Bacitracin 10 U: It is applied in a sputum culture plate (chocolate agar) to inhibit normal flora, including gram-positive bacteria, such as streptococci, and most species of Neisseria whereas to support the growth of Haemophilus which is resistant.
  • ONPG disks: The ONPG test is useful in differentiating members of the family Enterobacteriaceae and genera Neisseria based on b-D-galactosidase activity. ONPG stands for o-Nitrophenyl-b-D-galactopyranoside. Positive –Escherichia coli ATCC 25922 and Neisseria lactamica ATCC 23971 whereas negative Proteus mirabilis ATCC 12453 and Neisseria gonorrhoeae ATCC 43069.
  • X, V, and XV disks: For Haemophilus and its species identification.
  • Cefoxitin disk: It is used for screening Methicillin-Resistant Staphylococcal aureus (MRSA) and the Zone of inhibition (ZOI) should be less than 21 mm.

Haemophilus species Identification Using X, V, and XV Growth Factors Disks

Isolation of numerous Haemophilus species on the basis of growth characteristics are as follows-

SpeciesX DiskV diskCO2Hemolysis
on horse blood agar
Haemophilus
influenzae
++
H. parainfluenzae+
H. ducreyi+variablevariable
H. aegyptius++
H.haemolyticus+++
H. parahaemolyticus++
H. aphrophilus++
H. paraaphrophilus++
Table: Haemophilus species Identification Using X, V, and XV disks

Table: Haemophilus species Identification Using X, V, and XV Growth Factors Disks

Keynotes

  1. Haemophilus influenzae is xylose positive while Haemophilus aegyptius is negative.
  2. The x-factor is hemin and it is required for the synthesis of respiratory enzymes and is also called a heat-stable growth-promoting substance present in RBCs.
  3. V factor is nicotinamide-adenine dinucleotides (NAD)and it is applied as an electron carrier and is also known as a heat-labile vitamin-like substance present in red blood cells.
  4. Chocolate agar is a very useful medium to isolate fastidious organisms in Microbiology Laboratory from various clinical specimens like sputum ( H. influenzae), urethral discharge ( N. gonorrhoeae), CSF/blood (N. menigitidis).
  5. Nutrient agar is used for X, V, and XV factors assaying for Haemophilis species identification.

Related Footages

Novobiocin and Cefoxitin Resistant Staphylococcus saprophyticus

Fig. Novobiocin and Cefoxitin Resistant Staphylococcus saprophyticus

Optochin Susceptibility Test-Positive

Fig. Optochin Susceptibility Test-Positive Streptococcus pneumoniae

Oxidase Test-Positive and Negative Bacteria

Fig. Oxidase Test-Positive (left disk) and Negative (right disk) Bacteria

Bacitracin (O.O4U)-Sensitive Streptococcus pyogenes

Fig. Bacitracin (O.O4U)-Sensitive Streptococcus pyogenes

Haemophilus growth around 10 U bacitracin disk on chocolate agar of sputum culture

Fig. Haemophilus growth around 10 U bacitracin disk on chocolate agar of sputum culture

ONPG Test Negative and Positive Bacteria Demonstration

Fig. ONPG Test Negative and Positive Bacteria Demonstration

Haemophilus influenzae growth around XV disk growth factor and between X and V disks

Fig. Haemophilus influenzae growth around XV disk growth factor and between X and V disks

Haemophilus influenza growth on chocolate agar

Fig. Haemophilus influenzae colony morphology on chocolate agar

Haemophilus influenzae growth around XV disk growth factor on Nutrient Agar

Fig. Haemophilus influenzae growth around XV disk growth factor on Nutrient Agar

Haemophilus ducreyi growth around X disk and XV disk

Fig. Haemophilus ducreyi growth around X disk and XV disk

Haemophilus parainfluenzae growth around V disk and XV disk

Fig. Haemophilus parainfluenzae growth around V disk and XV disk

Cefoxitin Resistant Staphylococcus aureus

Fig. Cefoxitin Resistant Staphylococcus aureus (MRSA strain)


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