Introduction of a hydrolysis probe PCR assay for high-throughput screening of methicillin-resistant Staphylococcus aureus with the ability to include or exclude detection of Staphylococcus argenteus

Many countries using sensitive screening methods for detection of carriage of methicillin-resistant Staphylococcus aureus (MRSA) have a sustained low incidence of MRSA infections. For diagnostic laboratories with high sample volumes, MRSA screening requires stability, low maintenance and high perfor...

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Bibliographic Details
Published in:PloS one 2018, Vol.13 (2), p.e0192782-e0192782
Main Authors: Bogestam, Katja, Vondracek, Martin, Karlsson, Mattias, Fang, Hong, Giske, Christian G
Format: Article
Language:English
Subjects:
Antibiotics
Assaying
Biology and Life Sciences
Diagnostic systems
Drug resistance
Genes
Genomes
Genomics
High-throughput screening
Hospitals
Hydrolysis
Infectious diseases
Laboratories
Medical screening
Medicin och hälsovetenskap
Medicine and health sciences
Methicillin
Microbiology
Oligonucleotides
Pathogens
Physical Sciences
Research and Analysis Methods
Sensitivity
Staphylococcus argenteus
Staphylococcus aureus
Staphylococcus infections
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Summary:Many countries using sensitive screening methods for detection of carriage of methicillin-resistant Staphylococcus aureus (MRSA) have a sustained low incidence of MRSA infections. For diagnostic laboratories with high sample volumes, MRSA screening requires stability, low maintenance and high performance at a low cost. Herein we designed oligonucleotides for a new nuc targeted hydrolysis probe PCR to replace the standard in-house nuc SybrGreen PCR assay. This new, more time-efficient, PCR assay resulted in a 40% increase in daily sample capacity, with maintained high specificity and sensitivity. The assay was also able to detect Staphylococcus aureus clonal cluster 75 (CC75) lineage strains, recently re-classified as Staphylococcus argenteus, with a sensitivity considerably increased compared to our previous assay. While awaiting consensus if the CC75 lineage of S. aureus should be considered as S. argenteus, and whether methicillin-resistant S. argenteus should be included in the MRSA definition, many diagnostic laboratories need to update their MRSA assay sensitivity/specificity towards this lineage/species. The MRSA screening assay presented in this manuscript is comprised of nuc oligonucleotides separately targeting S. aureus and CC75 lineage strains/S. argenteus, thus providing high user flexibility for the detection of CC75 lineage strains/S. argenteus.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0192782