highly sensitive detection of staphylococcus aureus directly from patient blood高度敏感的直接从患者血液检测金黄色葡萄球菌.pdfVIP
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highly sensitive detection of staphylococcus aureus directly from patient blood高度敏感的直接从患者血液检测金黄色葡萄球菌
Highly Sensitive Detection of Staphylococcus aureus Directly from Patient Blood 1 1 2 2 Padmapriya P. Banada , Soumitesh Chakravorty , Darshini Shah , Michele Burday , Fermina M. 2 1 Mazzella , David Alland * 1 Division for Infectious Diseases, Department of Medicine, New Jersey Medical School – University of Medicine and Dentistry of New Jersey, Newark, New Jersey, United States of America, 2 Department of Pathology and Laboratory Medicine, New Jersey Medical School – University of Medicine and Dentistry of New Jersey, Newark, New Jersey, United States of America Abstract Background: Rapid detection of bloodstream infections (BSIs) can be lifesaving. We investigated the sample processing and assay parameters necessary for highly-sensitive detection of bloodstream bacteria, using Staphylococcus aureus as a model pathogen and an automated fluidic sample processing – polymerase chain reaction (PCR) platform as a model diagnostic system. Methodology/Principal Findings: We compared a short 128 bp amplicon hemi-nested PCR and a relatively shorter 79 bp amplicon nested PCR targeting the S. aureus nuc and sodA genes, respectively. The sodA nested assay showed an enhanced limit of detection (LOD) of 5 genomic copies per reaction or 10 colony forming units (CFU) per ml blood over 50 copies per reaction or 50 CFU/ml for the nuc assay. To establish optimal extraction protocols, we investigated the relative abundance of the bacteria in different components of the blood (white blood cells (WBCs), plasma or whole blood), using the above assays. The blood samples were obtained from the patients who were culture positive for S. aureus. Whole blood resulted in maximum PCR positives with sodA assay (90% positive) as opposed to cell
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