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Journal of Clinical Microbiology
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Mycology

Evaluation of the Amplex eazyplex Loop-Mediated Isothermal Amplification Assay for Rapid Diagnosis of Pneumocystis jirovecii Pneumonia

Timo Huber, Annerose Serr, Walter Geißdörfer, Christina Hess, Christian Lynker-Aßmus, Friederike D. von Loewenich, Christian Bogdan, Jürgen Held
Kimberly E. Hanson, Editor
Timo Huber
aMikrobiologisches Institut-Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen und Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
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Annerose Serr
bDepartment für Medizinische Mikrobiologie und Hygiene, Universitätsklinikum Freiburg, Freiburg, Germany
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Walter Geißdörfer
aMikrobiologisches Institut-Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen und Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
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Christina Hess
bDepartment für Medizinische Mikrobiologie und Hygiene, Universitätsklinikum Freiburg, Freiburg, Germany
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Christian Lynker-Aßmus
aMikrobiologisches Institut-Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen und Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
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Friederike D. von Loewenich
cDepartment of Medical Microbiology and Hygiene, Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany
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Christian Bogdan
aMikrobiologisches Institut-Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen und Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
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Jürgen Held
aMikrobiologisches Institut-Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen und Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
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Kimberly E. Hanson
University of Utah
Roles: Editor
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DOI: 10.1128/JCM.01739-20
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ABSTRACT

Quantitative PCR (qPCR) assays are the gold standard for diagnosis of Pneumocystis jirovecii pneumonia (PCP). However, they are laborious and require skilled personnel. Therefore, execution outside regular working hours of the molecular biology laboratory is limited. The eazyplex P. jirovecii assay (PJA) uses loop-mediated isothermal amplification for detection of P. jirovecii. It is performed directly with respiratory specimens, without the need for special skills, and delivers a result within 3 to 25 min. The goal of our study was to compare the performance of the eazyplex PJA with that of established P. jirovecii qPCR assays. All archived bronchoalveolar lavage fluid (BALF) samples that had previously tested positive for P. jirovecii by qPCR assay and 50 control samples (retrospective part), as well as all BALF samples received for P. jirovecii analysis over a period of 4 months (prospective part), were tested. Forty-nine patients with proven PCP and 126 patients without PCP were included. The sensitivity and specificity of the eazyplex PJA (95.7% and 96.5%, respectively) were comparable to those for three different P. jirovecii qPCR assays. The detection limit of the eazyplex PJA was analogous to 103 copies of the major surface glycoprotein gene per 25 μl of BALF, corresponding to 10 to 20 P. jirovecii cells. The eazyplex PJA reliably discriminated patients with PCP from patients with P. jirovecii colonization. It delivered a positive result within a mean of 9 min 38 s and required a hands-on time of 2 min 45 s. In summary, the eazyplex PJA showed identical performance for the diagnosis of PCP, compared to qPCR assays. However, in terms of time to result, practicability, and robustness, the eazyplex PJA is clearly superior and allows for around-the-clock molecular testing.

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Evaluation of the Amplex eazyplex Loop-Mediated Isothermal Amplification Assay for Rapid Diagnosis of Pneumocystis jirovecii Pneumonia
Timo Huber, Annerose Serr, Walter Geißdörfer, Christina Hess, Christian Lynker-Aßmus, Friederike D. von Loewenich, Christian Bogdan, Jürgen Held
Journal of Clinical Microbiology Nov 2020, 58 (12) e01739-20; DOI: 10.1128/JCM.01739-20

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Evaluation of the Amplex eazyplex Loop-Mediated Isothermal Amplification Assay for Rapid Diagnosis of Pneumocystis jirovecii Pneumonia
Timo Huber, Annerose Serr, Walter Geißdörfer, Christina Hess, Christian Lynker-Aßmus, Friederike D. von Loewenich, Christian Bogdan, Jürgen Held
Journal of Clinical Microbiology Nov 2020, 58 (12) e01739-20; DOI: 10.1128/JCM.01739-20
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KEYWORDS

LAMP
PCR
NAT
PCP
Pneumocystis carinii
HIV
BALF
AIDS
point-of-care

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