Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry and real-time PCR in a combined protocol for diagnosis of bloodstream infections: a turnaround time approach.

Autor:	Quiles MG; Universidade Federal de São Paulo - EPM/UNIFESP, Laboratório Especial de Microbiologia Clínica - Escola Paulista de Medicina, São Paulo, SP, Brazil. Electronic address: milenequiles@yahoo.com.br., Boettger BC; Universidade Federal de São Paulo - EPM/UNIFESP, Laboratório Especial de Microbiologia Clínica - Escola Paulista de Medicina, São Paulo, SP, Brazil., Inoue FM; Associação Fundo de Incentivo à Pesquisa, AFIP Medicina Diagnóstica, São Paulo, SP, Brazil., Monteiro J; Associação Fundo de Incentivo à Pesquisa, AFIP Medicina Diagnóstica, São Paulo, SP, Brazil., Santos DW; Hospital do Rim e Hipertensão - HRIM, São Paulo, SP, Brazil., Ponzio V; Hospital do Rim e Hipertensão - HRIM, São Paulo, SP, Brazil., Carlesse F; Instituto de Oncologia Pediátrica - GRAACC/UNIFESP, São Paulo, SP, Brazil., Cappellano P; Unidade de Hematologia e Transplante de Medula Óssea - UNIFESP, Hospital São Paulo, São Paulo, SP, Brazil., Carvalhaes CG; Hospital São Paulo, Laboratório Central - UNIFESP, São Paulo, SP, Brazil., Pignatari ACC; Universidade Federal de São Paulo - EPM/UNIFESP, Laboratório Especial de Microbiologia Clínica - Escola Paulista de Medicina, São Paulo, SP, Brazil.
Jazyk:	angličtina
Zdroj:	The Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases [Braz J Infect Dis] 2019 May - Jun; Vol. 23 (3), pp. 164-172. Date of Electronic Publication: 2019 Jun 05.
DOI:	10.1016/j.bjid.2019.05.005
Abstrakt:	Bloodstream infections (BSIs) are serious infections associated with high rates of morbidity and mortality. Every hour delay in initiation of an effective antibiotic increases mortality due to sepsis by 7%. Turnaround time (TAT) for conventional blood cultures takes 48h, forcing physicians to streamline therapy by exposing patients to broad-spectrum antimicrobials. Our objective was (1) to evaluate the accuracy and TAT of an optimized workflow combining direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and in-house real-time polymerase chain reaction (PCR) for bacterial identification and antimicrobial resistance profiling directly from positive blood bottles for diagnosing bloodstream infections and (2) to verify the effect of reporting results to medical staff. A total of 103 BSI episodes from 91 patients admitted to three hospitals in São Paulo, Brazil were included. TAT from molecular versus conventional methods was measured and compared. Our protocol showed an overall agreement of 93.5% for genus and 78.5% for species identification; 74.2% for methicillin resistance detection, 89.2% for extended-spectrum β-lactamase profiling, 77.8% for metallo-β-lactamase profiling, and 100% for carbapenemase profile and vancomycin-resistance detection when compared with conventional testing. TAT of molecular sample processing according to our protocol was 38h shorter than conventional methods. Antimicrobial interventions were possible in 27 BSI episodes. Antimicrobial discontinuation was achieved in 12 BSI episodes while escalation of therapy occurred in 15 episodes. Antimicrobial therapy was inadequate in three (12%) BSI episodes diagnosed using results of molecular testing. Our in-house rapid protocol for identifying both bacteria and antimicrobial resistance provided rapid and accurate results, having good agreement with conventional testing results. These results could contribute to faster antimicrobial therapy interventions in BSI episodes. (Copyright © 2019 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U. All rights reserved.)
Databáze:	MEDLINE
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