title: Evaluation of a Commercial Multiplex PCR Assay for Detection of Pathogen DNA in Blood from Patients with Suspected Sepsis.
abstract: The Magicplex Sepsis Real-time Test (MST) is a commercial multiplex PCR that can detect more than 90 different pathogens in blood, with an analysis time of six hours. The aim of the present study was to evaluate this method for the detection of bloodstream infection (BSI). An EDTA whole blood sample for MST was collected together with blood cultures (BC) from patients with suspected sepsis at the Emergency Department of a university hospital. Among 696 study patients, 322 (46%) patients were positive with at least one method; 128 (18%) were BC positive and 268 (38%) were MST positive. Considering BC to be the gold standard, MST had an overall sensitivity of 47%, specificity of 66%, positive predictive value (PPV) of 23%, and a negative predictive value of 87%. Among the MST positive samples with a negative BC, coagulase-negative staphylococci (CoNS) and species that rarely cause community-acquired BSI were frequently noted. However, the quantification cycle (Cq) values of the MST+/BC- results were often high. We thus hypothesized that the performance of the MST test could be improved if the Cq cut-off level was adjusted downwards. With a lower Cq cut-off value, i.e. 6.0 for Staphylococcus species and 9.0 for all other species, the number of MST positive cases decreased to 83 (12%) and the overall sensitivity decreased to 38%. However, the PPV increased to 59% and the specificity increased to 96%, as many MST positive results for CoNS and bacteria that rarely cause community-acquired BSI turned MST negative. In conclusion, our study shows that with a lower Cq cut-off value, the MST will detect less contaminants and findings with unclear relevance, but to the cost of a lower sensitivity. Consequently, we consider that a positive MST results with a Cq value above the adjusted cut-off should be interpreted with caution, as the result might be clinically irrelevant. In a correspondent way, quantitative results could probably be useful in the interpretation of positive results from other molecular assays for the detection of BSI.
authors.affiliation: Department of Infectious Diseases, Örebro University Hospital, Örebro, Sweden.|School of Health and Medical Sciences, Örebro University, Örebro, Sweden.
authors.forename: Ingrid
authors.identifier: http://orcid.org/0000-0002-5929-212X
authors.initials: I
authors.lastname: Ziegler
authors.affiliation: School of Health and Medical Sciences, Örebro University, Örebro, Sweden.|Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Fagerström
authors.affiliation: School of Health and Medical Sciences, Örebro University, Örebro, Sweden.|Department of Infectious Diseases, Karolinska University Hospital, Stockholm, Sweden.|Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Strålin
authors.affiliation: School of Health and Medical Sciences, Örebro University, Örebro, Sweden.|Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
authors.forename: Paula
authors.initials: P
authors.lastname: Mölling
mesh_terms: D004269:DNA, Bacterial / Q000097:blood / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D018805:Sepsis / Q000097:blood / Q000235:genetics / Q000382:microbiology; D013203:Staphylococcal Infections / Q000097:blood / Q000235:genetics / Q000382:microbiology; D013210:Staphylococcus / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial