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Detection of bacteraemia in critically ill patients using 16S rDNA polymerase chain reaction and DNA sequencing
J Sleigh1, R Cursons, M La Pine
1Department of Intensive Care, Waikato Hospital, Pembroke Street, Hamilton, New Zealand. sleighj@hwl.co.nz
Objective:
To confirm the sensitivity of the polymerase chain reaction (PCR) technique (versus blood cultures) and to gain a better understanding of the incidence of true- and false-positive results when using this technique.
Design:
Observational study.
Setting:
Fourteen-bed, level 3 intensive care unit.
Patients:
Hundred twenty-six critically ill adult patients. Hundred ninety-seven blood culture and PCR samples taken as clinically indicated for suspected sepsis, according to routine ICU protocol.
Measurements And Results:
The PCR product (16SrDNA: 341F-1195R) was sequenced and compared with a database of known species (Genebank) to identify the bacterial nucleic acid. The PCR or blood culture result was classified as a true-positive if there was other microbiological or clinical supporting evidence.