Central Philippine University

College of Medical Laboratory Science

MLS 2309 Clinical Bacteriology

Activity No. 5
LABORATORY DISCUSSION

Answer the following questions comprehensively. You may use either written paragraph or bullet-point
format. Your answer(s) will be graded based on the rubrics below.

1. In the coagulase test, two methods were employed, the Slide and Tube coagulase test. Differentiate
the type of coagulase that are detected by the two methods in terms of its activity and action.

Slide coagulase test is done to detect “bound coagulase” or “clumping factor”. Clumping factor
causes clumping by acting directly on the fibrinogen in the plasma. Some strains will yield a negative
slide coagulase test because the clumping components are not present in all S. aureus. Furthermore,
cell surface capsular polysaccharides can hide clumping cause. As a result, the tube coagulase test
must be used to validate the negative slide tests. Other Staphylococcus species, such as S.
lugdunensis and S. schleiferi, can result in positive slide coagulase test findings. While a tube
coagulase test detects free coagulase (staphylocoagulase) that interacts with coagulase-reacting
factor, a coagulase-reacting factor test detects coagulase-reacting factor (CRF). CRF is a molecule
that resembles thrombin. A positive test is indicated by the development of a clot within four hours.
Indirect fibrinogen to fibrin conversion occurs when staphylocoagulase and CRF work together. S.
aureus fibrinolysin or staphylokinase, a plasmid-carried enzyme that is more active at 35°C than at
25°C, can dissolve the coagulase clot that has formed. As a result, the tube coagulase tubes must
be checked for clot development every 1, 2, and 4 hours. Negative tubes should be kept at room
temperature overnight.

Slide coagulase test is prepared by emulsify a staphylococcal colony in a drop of water on a clean
and grease-free glass slide with a minimum of spreading (If the isolate does not form a smooth, milky
suspension, do not proceed with the test). While, Tube coagulase test is prepare a 1-in-6 dilution of
the plasma in saline (0.85% NaCl) and place 1 ml volumes of the diluted plasma in small tubes.

In slide coagulase test will yield a coagulase positive if macroscopic clumping in 10 seconds or less in
coagulated plasma drop and no clumping in saline or water drop and coagulase negative if no
clumping in either drop. While in the tube coagulase test it will yield a result of coagulase positive if
there is a presence of clot of any size eg. Staphylococcus aureus and a coagulase negative if no
clot (plasma remains wholly liquid or shows only a flocculent or ropy precipitate).
eg. Staphylococcus epidermidis.

2. Is the Cystine tryptic agar (CTA) carbohydrate test still recommended for the detection of acid
production of Neisseria species? Support your answer.

The conventional test for identifying acid generation by Neisseria and related species was the CTA-
carbohydrate test, however it is no longer recommended for detecting acid generated by Neisseria
species. Fermentative organisms create more acid from carbohydrates than oxidative Neisseria spp.,
hence CTA-carbohydrate media are intended to detect acid generation from them. On CTA-
carbohydrate media, it is also expected that organisms generate acid as they develop; however,
strains of Neisseria spp., including N. gonorrhoeae, do not grow well in these media. It may be
difficult to acquire reliable findings since the percentage of peptone to carbohydrate in CTA
medium is high, and Neisseria spp. have generated ammonia from peptone and neutralized the
acid produced by carbs. As a result, alternative media have been created that either have a lower
peptone to carbohydrate ratio or employ carbohydrate solutions injected with heavy suspensions
and rely on preformed enzymes present in the suspension to make acid.
 Central Philippine University
College of Medical Laboratory Science
MLS 2309 Clinical Bacteriology

References:

Tankeshwar, Acharya. “Coagulase Test: Principle, Procedure, Results • Microbe Online.” Microbe Online, 2
July 2021, microbeonline.com/diagnostic-tests-biochemical-tests-coagulase-test/.

“Acid Detection Test - Gonorrhea - STD Information from CDC.” Centers for Disease Control and
Prevention, Centers for Disease Control and Prevention, 10 Dec. 2013,
www.cdc.gov/std/gonorrhea/lab/tests/acid.htm.