Diagnosis by real-time polymerase chain reaction of pathogens and antimicrobial resistance genes in bone marrow transplant patients with bloodstream infections

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Menezes, Liana Carballo [UNIFESP]
Rocchetti, Talita Trevizani [UNIFESP]
Bauab, Karen de Castro [UNIFESP]
Cappellano, Paola [UNIFESP]
Quiles, Milene Goncalves [UNIFESP]
Carlesse, Fabianne [UNIFESP]
Oliveira, José Salvador Rodrigues de [UNIFESP]
Pignatari, Antonio Carlos Campos [UNIFESP]
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Background: Early identification of pathogens and antimicrobial resistance in bloodstream infections (BSIs) decreases morbidity and mortality, particularly in immunocompromised patients. the aim of the present study was to compare real-time polymerase chain reaction (PCR) with commercial kits for detection of 17 pathogens from blood culture (BC) and 10 antimicrobial resistance genes.Methods: A total of 160 BCs were taken from bone marrow transplant patients and screened with Gram-specific probes by multiplex real-time PCR and 17 genus-specific sequences using TaqMan probes and blaSHV, blaTEM, blaCTX, blaKPC, blaIMP, blaSPM, blaVIM, vanA, vanB, and mecA genes by SYBR Green.Results: Twenty-three of 33 samples identified by phenotypic testing were concordantly positive by BC and real-time PCR. Pathogen identification was discordant in 13 cases. in 12 of 15 coagulase-negative staphylococci, the mecA gene was detected and four Enterococcus spp. were positive for vanA. Two blaCTX and three blaSHV genes were found by quantitative PCR. the blaKPC and metallo-beta-lactamase genes were not detected. Five fungal species were identified only by real-time PCR.Conclusions: Real-time PCR could be a valuable complementary tool in the management of BSI in bone marrow transplants patients, allowing identification of pathogens and antimicrobial resistance genes.
Bmc Infectious Diseases. London: Biomed Central Ltd, v. 13, 7 p., 2013.