A simple and expedient PCR format for rapid molecular screening of methicillin-resistant Staphylococcus aureus in Amies gel swabs.

Autor: McIver CJ; Department of Microbiology (NSW HP), St George Hospital, Sydney, NSW, Australia; School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia. Electronic address: christopher.mciver@health.nsw.gov.au., De Silva RND; Ministry of Health, Colombo, Sri Lanka., Er N; Department of Microbiology (NSW HP), St George Hospital, Sydney, NSW, Australia., Pratama R; Department of Microbiology (NSW HP), St George Hospital, Sydney, NSW, Australia., Mukerjee C; Department of Microbiology (NSW HP), St George Hospital, Sydney, NSW, Australia., Stevens R; Department of Microbiology (NSW HP), St George Hospital, Sydney, NSW, Australia; School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia., Taylor PC; Department of Microbiology (NSW HP), St George Hospital, Sydney, NSW, Australia; School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
Jazyk: angličtina
Zdroj: Pathology [Pathology] 2020 Jun; Vol. 52 (4), pp. 466-472. Date of Electronic Publication: 2020 Apr 10.
DOI: 10.1016/j.pathol.2020.02.005
Abstrakt: Screening patients for carriage of methicillin-resistant Staphylococcus aureus (MRSA) is commonly undertaken in hospital laboratories using phenotypic methods. This work is labour-intensive, costly and may take several days to complete. We report on the validation of a novel rapid screening approach for direct testing of Amies gel swabs for MRSA. The method is based on two quantitative real time-PCR (qRT-PCR) assays for the detection of the nuc and mecA genes of MRSA. Based on SYBR Green technology, the assays use significantly less reagents than conventional qRT-PCR methods and are applied to testing templates derived directly from aqueous suspensions of swabs. Notwithstanding the occurrence of false-positives due to non-specific fluorescence generated by the SYBR Green dye, the novel assays showed a high negative predictive value enabling earlier reporting of negative findings and selection of swabs for confirmatory phenotypic testing for MRSA. In a blinded trial of 461 swabs, of which 34 (7.4%) were previously shown to be culture-positive for MRSA, the novel assays selected 121 (26.2%) swabs (inclusive of the known MRSA-positive swabs) for phenotypic testing. This enabled early reporting of negative findings for 340 (73.8%) of the 461 swabs tested. Application of this method has implications for screening strategies for large laboratories whilst achieving cost benefits.
(Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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