Gram-positive isolates from blood
16 min
- Tell S. aureus from S. lugdunensis and other staphylococci in blood
- Tell streptococci from enterococci and their vancomycin-resistant look-alikes in blood
- Recognize a catalase-positive blood rod that needs a Listeria workup
Try first
Get the idea
Gram stain and catalase first
The positive bottle's Gram stain decides where identification starts, and catalase splits the gram-positive organisms into groups. Take the colony for catalase without any blood agar, because red cells contain catalase and give a false-positive reaction.1
| Gram stain and colony | Catalase | Group |
|---|---|---|
| Cocci in clusters | Positive | Staphylococcus |
| Cocci in pairs or chains | Negative | Streptococcus, Enterococcus and look-alikes |
| Club-shaped or palisading rods | Usually positive | Corynebacterium and other coryneforms |
| Small rods or coccobacilli, narrow beta hemolysis | Positive | Listeria monocytogenes |
Staphylococci
The slide coagulase test detects clumping factor. The tube test detects free coagulase. S. lugdunensis can be slide positive and tube negative, and it is more invasive than most coagulase-negative staphylococci.1 S. aureus in blood is usually significant, even in one set. A validated system gives the final species.1
Catalase-negative cocci
Hemolysis, Lancefield group and species identification sort these cocci.1
- Enterococcus is pyrrolidonyl arylamidase (PYR) positive and bile-esculin positive, and most strains grow in 6.5% NaCl.
- The S. gallolyticus group is often group D and bile-esculin positive, is PYR negative, and does not grow in 6.5% NaCl. S. gallolyticus subsp. gallolyticus bacteremia is strongly linked to endocarditis and colorectal neoplasia, so the species name matters.2
- Leuconostoc and Pediococcus are intrinsically vancomycin resistant. A vancomycin-resistant catalase-negative coccus needs species confirmation before it is reported as an enterococcus.1
Coryneform rods and Listeria
A single set growing a coryneform rod commonly reflects skin contamination.1 Listeria monocytogenes forms small rods or coccobacilli with a narrow zone of beta hemolysis, so it can pass for a diphtheroid or group B streptococcus. The Gram stain shows a rod, and a positive catalase separates it from group B streptococcus. Tumbling motility at room temperature, umbrella motility in semisolid medium and a validated identification system complete the workup.1,3 A Listeria isolate from blood or CSF is significant.1
References
- Carroll KC, Pfaller MA, Landry ML, McAdam AJ, Karlowsky JA, Patel R, Pritt BS, eds. Manual of Clinical Microbiology. 13th ed. ASM Press; 2023.
- Putnam NE, Youn J, Wallace MA, et al. Comparative evaluation of current biochemical-, sequencing-, and proteomic-based identification methods for the Streptococcus bovis group. J Clin Microbiol. 2023;61(4):e01712-22. doi:10.1128/jcm.01712-22
- Centers for Disease Control and Prevention. Listeria monocytogenes identification chart. Accessed September 27, 2026. https://reach.cdc.gov/sites/default/files/job-aids-resources/Listeria_monocytogenes_ID_Chart_Branded_508.pdf
Watch one
A blood bottle grows gram-positive cocci in pairs and short chains. The colonies are small and nonhemolytic, and catalase is negative. The susceptibility panel reports a vancomycin MIC of 64 µg/mL. A colleague wants to report vancomycin-resistant Enterococcus (VRE).
What do you do before reporting?
- Read the Gram stain and catalase: catalase-negative cocci in pairs and chains fit streptococci, enterococci and their look-alikes.
Catalase and arrangement place the isolate among the catalase-negative cocci.
- Weigh the vancomycin result: it fits VRE, and it also fits Leuconostoc or Pediococcus.
An intrinsically resistant organism gives the same high MIC as acquired resistance.
- Ask for species evidence: the pattern so far gives no species, so send the colony for identification by MALDI-TOF or another validated system.
A VRE report puts the patient on contact precautions, so the species has to be right.
- Read the result: MALDI-TOF identifies Leuconostoc mesenteroides.
The confirmed species decides what the resistance means.
Your turn
Use it
- Priya Raman, 31, is 30 weeks pregnant with fever and muscle aches.
- Two blood-culture sets were drawn, and both aerobic bottles signal positive at 18 hours.
- The Gram stain shows small gram-positive rods and coccobacilli.
- On blood agar the colonies are small with a narrow zone of beta hemolysis.
The clue that settled this case is the positive catalase on a small rod with narrow beta hemolysis. It separated Listeria from group B streptococcus, and the tumbling motility separated it from a diphtheroid. MALDI-TOF confirmed L. monocytogenes from both sets.
Results
- Tell S. aureus from S. lugdunensis and other staphylococci in blood
- Tell streptococci from enterococci and their vancomycin-resistant look-alikes in blood
- Recognize a catalase-positive blood rod that needs a Listeria workup
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