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

BMC Infect Dis. 2013 Apr 5:13:166. doi: 10.1186/1471-2334-13-166.

Abstract

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-β-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Bacteria / classification
  • Bacteria / drug effects
  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Bone Marrow Transplantation / adverse effects
  • Child
  • Drug Resistance, Bacterial*
  • Drug Resistance, Fungal*
  • Fungi / classification
  • Fungi / drug effects
  • Fungi / genetics
  • Fungi / isolation & purification*
  • Humans
  • Immunocompromised Host
  • Microbiological Techniques / methods*
  • Multiplex Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / methods*
  • Sepsis / microbiology*