Multiplex PCR assay underreports true bloodstream infections with coagulase-negative staphylococci in hematological patients with febrile neutropenia

2016 ◽  
Vol 85 (4) ◽  
pp. 413-415 ◽  
Author(s):  
Yvonne Reers ◽  
Evgeny A. Idelevich ◽  
Hanna Pätkau ◽  
Maria Cristina Sauerland ◽  
Sascha Tafelski ◽  
...  
2018 ◽  
Vol 5 (10) ◽  
Author(s):  
Brian A Buss ◽  
Timothy J Baures ◽  
Minkyoung Yoo ◽  
Kimberly E Hanson ◽  
Donald P Alexander ◽  
...  

Abstract Implementation of Biofire FilmArray Blood Culture Identification Multiplex PCR panel (BCID) at a cancer hospital was associated with reduced time to appropriate antimicrobial therapy. Additional reductions were not observed when BCID was coupled with antimicrobial stewardship intervention.


2017 ◽  
Vol 55 (6) ◽  
pp. 1857-1864 ◽  
Author(s):  
Jo-Ann McClure ◽  
Johanna Zaal DeLongchamp ◽  
John M. Conly ◽  
Kunyan Zhang

ABSTRACTMethicillin-resistantStaphylococcus aureus(MRSA) is a clinically significant pathogen that is resistant to a wide variety of antibiotics and responsible for a large number of nosocomial infections worldwide. The Agency for Healthcare Research and Quality and the Centers for Disease Control and Prevention recently recommended the adoption of universal mupirocin-chlorhexidine decolonization of all admitted intensive care unit patients rather than MRSA screening with targeted treatments, which raises a serious concern about the selection of resistance to mupirocin and chlorhexidine in strains of staphylococci. Thus, a simple, rapid, and reliable approach is paramount in monitoring the prevalence of resistance to these agents. We developed a simple multiplex PCR assay capable of screeningStaphylococcusisolates for the presence of antiseptic resistance genes for chlorhexidine and quaternary ammonium compounds, as well as mupirocin and methicillin resistance genes, while simultaneously discriminatingS. aureusfrom coagulase-negative staphylococci (CoNS). The assay incorporates 7 PCR targets, including theStaphylococcus16S rRNA gene (specifically detectingStaphylococcusspp.),nuc(distinguishingS. aureusfrom CoNS),mecA(distinguishing MRSA from methicillin-susceptibleS. aureus),mupAandmupB(identifying high-level mupirocin resistance), andqacandsmr(identifying chlorhexidine and quaternary ammonium resistance). Our assay demonstrated 100% sensitivity, specificity, and accuracy in a total of 23 variant antiseptic- and/or antibiotic-resistant control strains. Further validation of our assay using 378 randomly selected and previously well-characterized local clinical isolates confirmed its feasibility and practicality. This may prove to be a useful tool for multidrug-resistantStaphylococcusmonitoring in clinical laboratories, particularly in the wake of increased chlorhexidine and mupirocin treatments.


2013 ◽  
Vol 62 (11) ◽  
pp. 1673-1679 ◽  
Author(s):  
Lucrecia Carrara ◽  
Ferran Navarro ◽  
Miquel Turbau ◽  
Montse Seres ◽  
Indalecio Morán ◽  
...  

Mortality from bloodstream infections (BSIs) correlates with diagnostic delay and the use of inappropriate empirical treatment. Early PCR-based diagnosis could decrease inappropriate treatment, improving patient outcome. The aim of the present study was to assess the clinical utility of this molecular technology to diagnose BSIs. We assessed a new dual-priming oligonucleotide-based multiplex PCR assay, the Magicplex Sepsis Test (MST) (Seegene), along with blood culture (BC). A total of 267 patients from the intensive care unit and haematology and emergency departments were enrolled. Clinical data were also used by physicians to determine the likelihood of infection. Ninety-eight (37 %) specimens were positive: 29 (11 %) by both the MST and BC, 29 (11 %) by the MST only, and 40 (15 %) by BC only. The proportion of agreement between the two methods was 73 % (Cohen’s κ: 0.45; 0.28–0.6; indicating fair to moderate agreement). According to clinical assessment, 63 (64 %) positive specimens were considered BSIs: 23 (36 %) were positive by both the MST and BC, 22 (35 %) were positive only by BC, and 18 (29 %) were positive only by the MST. Thirty-eight (14 %) positive specimens by the MST and/or BC were considered as contaminants. Of 101 specimens collected from patients receiving antibiotics, 20 (20 %) were positive by the MST and 32 (32 %) by BC. Sensitivity and specificity were 65 % and 92 %, respectively, for the MST and 71 % and 88 %, respectively for BC. We concluded that the MST shows a high specificity but changes in design are needed to increase bacteraemia detection. For viability in clinical laboratories, technical improvements are also required to further automate the process.


2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42


Author(s):  
Sunarno ◽  
Khariri ◽  
Fauzul Muna ◽  
Kambang Sariadji ◽  
Yuni Rukminiati ◽  
...  

2010 ◽  
Vol 105 (2) ◽  
pp. 151-155 ◽  
Author(s):  
Mollah Md. Hamiduzzaman ◽  
Ernesto Guzman-Novoa ◽  
Paul H. Goodwin

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