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Bacteriology

A Test for the Rapid Detection of the Cefazolin Inoculum Effect in Methicillin-Susceptible Staphylococcus aureus

Sandra Rincon, Lina P. Carvajal, Sara I. Gomez-Villegas, Aura M. Echeverri, Rafael Rios, An Dinh, Claudia Pedroza, Karen M. Ordoñez, Esteban Nannini, Zhizeng Sun, Vance G. Fowler, Barbara E. Murray, William R. Miller, Timothy Palzkill, Lorena Diaz, Cesar A. Arias, Jinnethe Reyes
Carey-Ann D. Burnham, Editor
Sandra Rincon
aMolecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia
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Lina P. Carvajal
aMolecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia
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Sara I. Gomez-Villegas
bCenter for Antimicrobial Resistance and Microbial Genomics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
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Aura M. Echeverri
aMolecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia
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Rafael Rios
aMolecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia
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An Dinh
bCenter for Antimicrobial Resistance and Microbial Genomics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
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Claudia Pedroza
cCenter for Clinical Research and Evidence-Based Medicine, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
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Karen M. Ordoñez
dE.S.E. Hospital Universitario San Jorge de Pereira, Risaralda, Colombia
eUniversidad Tecnológica de Pereira, Risaralda, Colombia
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Esteban Nannini
fFacultad de Ciencias Médicas, Universidad Nacional de Rosario, Rosario, Argentina
gInstituto de Inmunología Clínica y Experimental de Rosario, CONICET, Rosario, Argentina
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Zhizeng Sun
hBaylor College of Medicine, Houston, Texas, USA
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Vance G. Fowler
iDivision of Infectious Diseases, Department of Medicine, Duke University, Durham, North Carolina, USA
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Barbara E. Murray
bCenter for Antimicrobial Resistance and Microbial Genomics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
jDivision of Infectious Diseases, Department of Internal Medicine, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
kDepartment of Microbiology and Molecular Genetics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
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William R. Miller
bCenter for Antimicrobial Resistance and Microbial Genomics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
jDivision of Infectious Diseases, Department of Internal Medicine, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
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Timothy Palzkill
hBaylor College of Medicine, Houston, Texas, USA
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Lorena Diaz
aMolecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia
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Cesar A. Arias
bCenter for Antimicrobial Resistance and Microbial Genomics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
jDivision of Infectious Diseases, Department of Internal Medicine, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
kDepartment of Microbiology and Molecular Genetics, McGovern Medical School, University of Texas Health Science Center, Houston, Texas, USA
lCenter for Infectious Diseases, School of Public Health, University of Texas Health Science Center at Houston, Houston, Texas, USA
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Jinnethe Reyes
aMolecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia
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Carey-Ann D. Burnham
Washington University School of Medicine
Roles: Editor
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DOI: 10.1128/JCM.01938-20
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ABSTRACT

The cefazolin inoculum effect (CzIE) has been associated with therapeutic failures and mortality in invasive methicillin-susceptible Staphylococcus aureus (MSSA) infections. A diagnostic test to detect the CzIE is not currently available. We developed a rapid (∼3 h) CzIE colorimetric test to detect staphylococcal-β-lactamase (BlaZ) activity in supernatants after ampicillin induction. The test was validated using 689 bloodstream MSSA isolates recovered from Latin America and the United States. The cefazolin MIC determination at a high inoculum (107 CFU/ml) was used as a reference standard (cutoff ≥16 μg/ml). All isolates underwent genome sequencing. A total of 257 (37.3%) of MSSA isolates exhibited the CzIE by the reference standard method. The overall sensitivity and specificity of the colorimetric test was 82.5% and 88.9%, respectively. Sensitivity in MSSA isolates harboring type A BlaZ (the most efficient enzyme against cefazolin) was 92.7% with a specificity of 87.8%. The performance of the test was lower against type B and C enzymes (sensitivities of 53.3% and 72.3%, respectively). When the reference value was set to ≥32 μg/ml, the sensitivity for isolates carrying type A enzymes was 98.2%. Specificity was 100% for MSSA lacking blaZ. The overall negative predictive value ranged from 81.4% to 95.6% in Latin American countries using published prevalence rates of the CzIE. MSSA isolates from the United States were genetically diverse, with no distinguishing genomic differences from Latin American MSSA, distributed among 18 sequence types. A novel test can readily identify most MSSA isolates exhibiting the CzIE, particularly those carrying type A BlaZ. In contrast to the MIC determination using high inoculum, the rapid test is inexpensive, feasible, and easy to perform. After minor validation steps, it could be incorporated into the routine clinical laboratory workflow.

FOOTNOTES

    • Received 25 July 2020.
    • Returned for modification 20 September 2020.
    • Accepted 23 January 2021.
    • Accepted manuscript posted online 3 February 2021.
  • Supplemental material is available online only.

  • Copyright © 2021 American Society for Microbiology.

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A Test for the Rapid Detection of the Cefazolin Inoculum Effect in Methicillin-Susceptible Staphylococcus aureus
Sandra Rincon, Lina P. Carvajal, Sara I. Gomez-Villegas, Aura M. Echeverri, Rafael Rios, An Dinh, Claudia Pedroza, Karen M. Ordoñez, Esteban Nannini, Zhizeng Sun, Vance G. Fowler, Barbara E. Murray, William R. Miller, Timothy Palzkill, Lorena Diaz, Cesar A. Arias, Jinnethe Reyes
Journal of Clinical Microbiology Mar 2021, 59 (4) e01938-20; DOI: 10.1128/JCM.01938-20

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A Test for the Rapid Detection of the Cefazolin Inoculum Effect in Methicillin-Susceptible Staphylococcus aureus
Sandra Rincon, Lina P. Carvajal, Sara I. Gomez-Villegas, Aura M. Echeverri, Rafael Rios, An Dinh, Claudia Pedroza, Karen M. Ordoñez, Esteban Nannini, Zhizeng Sun, Vance G. Fowler, Barbara E. Murray, William R. Miller, Timothy Palzkill, Lorena Diaz, Cesar A. Arias, Jinnethe Reyes
Journal of Clinical Microbiology Mar 2021, 59 (4) e01938-20; DOI: 10.1128/JCM.01938-20
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KEYWORDS

CzIE rapid test
BlaZ
cefazolin inoculum effect
high inoculum

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