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Chapter 18 : General Approaches to Identification of Aerobic Gram-Positive Cocci

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Abstract:

Majority of aerobic, or facultatively aerobic, gram-positive cocci isolated from clinical specimens are distributed among the genera , , and . This chapter provides tables containing organisms with similar cellular morphologies, either "streptococcal," consisting of gram-positive cocci or coccobacilli arranged primarily in pairs and/or chains, or "staphylococcal," signifying that cells appear as cocci arranged in pairs, tetrads, clusters, and irregular groups. The commonly isolated aerobic gram-positive cocci (, , enterococci) can usually be accurately identified by determining a few basic phenotypic traits (cellular morphology, catalase reaction, and production of pyrrolidonyl arylamidase [PYR]). The chapter highlights the fact that it is increasingly difficult to identify some of the less frequently isolated organisms solely on the basis of phenotypic traits, as new genera and species of aerobic gram-positive cocci are described and characterized. Basic phenotypic tests can usually suggest a possible identity for strains of infrequently encountered aerobic gram-positive cocci, but evaluation with a larger battery of phenotypic tests or molecular identification methods is often valuable, if not indispensible, for accurate identification. Nucleic acid probe tests and amplification methods for identification of some of the commonly isolated aerobic gram-positive cocci are commercially available and designed for use in medium to large-volume clinical microbiology laboratories. The chapter concludes by emphasizing that the comparison of 16S rRNA gene sequences is the most useful method for molecular characterization of the aerobic gram-positive cocci of clinical interest, although sequence comparison of other genes may also be helpful for identification.

Citation: Ruoff K. 2011. General Approaches to Identification of Aerobic Gram-Positive Cocci, p 304-307. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch18

Key Concept Ranking

Gram-Positive Cocci
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Gram-Positive Coccobacilli
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Staphylococcus aureus
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Streptococcus pneumoniae
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Staphylococcus aureus
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Streptococcus pneumoniae
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References

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8. Fihman, V.,, L. Raskine,, Z. Barrou,, C. Kiffel,, J. Riahi,, B. Bercot,, and M.-J. Sanson-Le Pors. 2006. Lactococcus garvieae endocarditis: identification by 16S rRNA and sodA sequence analysis. J. Infect. 52:e3e6.
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18. Poyart, C.,, G. Quesne,, C. Boumaila,, and P. Trieu-Cuot. 2001. Rapid and accurate species-level identification of coagulase-negative staphylococci by using the sodA gene as a target. J. Clin. Microbiol. 39:42964301.
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Tables

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TABLE 1

Characteristics of catalase-negative gram-positive cocci that grow aerobically and form cells arranged in pairs and chains

See chapters 17 and 20 to 22 for methods for tests referred to in this table. Reactions shown are typical, but exceptions may occur. Abbreviations and symbols: PYR, production of pyrrolidonyl arylamidase; LAP, production of leucine aminopeptidase; 6.5% NaCl, growth in 6.5% NaCl; BE, hydrolysis of esculin in the presence of 40% bile; 45°C, growth at 45°C; probe, reaction with commercially available nucleic acid probe for the genus Enterococcus; HIP, hydrolysis of hippurate; satellitism, satellite growth behavior; 10°C, growth at 10°C; +, most strains positive; –, most strains negative; V, variable reactions are observed; NA, not applicable.

The reactions in this table are typical for , , and . cells tend to be arranged in clusters, and isolates are hippurate hydrolysis negative ( Table 2 ).

, , and cells tend to be arranged in pairs and chains, in contrast to the cells of , which are arranged in pairs, tetrads, and clusters ( Table 2 ).

is distinguished from the other catalase-negative organisms in Table 1 by its ability to produce gas as an end product of glucose metabolism and intrinsic resistance to vancomycin. The phenotypically similar genus contains organisms formerly classified as leuconostocs and the species formerly named (see chapter 22).

Most streptococci are PYR negative, with the exception of isolates and some strains of , which are PYR positive.

Citation: Ruoff K. 2011. General Approaches to Identification of Aerobic Gram-Positive Cocci, p 304-307. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch18
Generic image for table
TABLE 2

Differentiating features of gram-positive cocci that grow aerobically and form cells arranged in clusters or irregular groups

See chapters 17, 19, and 22 for methods for performing the phenotypic tests referred to in this table. Reactions shown are typical; exceptions may occur. Abbreviations: PYR, production of pyrrolidonyl arylamidase; LAP, production of leucine aminopeptidase; NaCl, growth in the presence of either 5% or 6.5% NaCl (see footnotes and ); ESC, esculin hydrolysis; BGUR, production of β-glucuronidase; +, most strains positive; –, most strains negative; V, variable reactions are observed; NA, not applicable; α, alpha-hemolysis on sheep blood agar; γ, nonhemolytic reaction on sheep blood agar; S, susceptible; R, resistant.

Growth in the presence of 5% sodium chloride.

, a related genus infrequently isolated from clinical specimens, is distinguished from by its ability to produce acid aerobically from D-glucose and glycerol (see chapter 19).

Growth in the presence of 6.5% sodium chloride.

isolates are usually catalase negative or weakly positive but may be strongly catalase positive.

( Table 1 ) exhibits reactions identical to those of in the PYR, ESC, and NaCl tests. However, cells are arranged primarily in chains, while cells usually form clusters. Other species form cells arranged in pairs and chains ( Table 1 ).

cells tend to be arranged in pairs, tetrads, and groups, in contrast to the cells of other species, which usually occur in pairs and short chains ( Table 1 ).

strains form tiny pinpoint nonhemolytic colonies on blood agar after 24 h of aerobic incubation at 35°C, while isolates form larger alpha-hemolytic colonies under similar incubation conditions. In contrast to , prefers aerobic incubation atmospheres. Two additional species of isolated from human sources have been described, each based on a single isolate. In contrast to , the new species and the proposed species “” are PYR negative (see chapter 22).

The genera and have similar phenotypic characteristics, except that tetragenococci are vancomycin susceptible. The bile esculin test can differentiate between tetragenococci (positive) and (negative) (see chapter 22).

Citation: Ruoff K. 2011. General Approaches to Identification of Aerobic Gram-Positive Cocci, p 304-307. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch18

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