Evaluation of buffy coat 16S rRNA PCR, buffy coat culture and whole blood PCR for detection of bacteraemia

College

College of Science

Department/Unit

Biology

Document Type

Article

Source Title

Memorias do Instituto Oswaldo Cruz

Volume

105

Issue

2

First Page

117

Last Page

122

Publication Date

1-1-2010

Abstract

The use of Gram type-specific PCR on buffy coat from clinical specimens for the detection of bacteraemia was evaluated for the first time using whole blood culture as the gold standard. In addition, the established buffy coat culture and whole blood PCR were also compared. Gram-positive bacteria belonging to six species and Gramnegative bacteria from 10 species were isolated and identified by culture and detected using broad-range 16S rDNA primers and Gram-specific primers. Data from the three methods all conferred very high sensitivity, specificity, positive and negative predictive values when compared to whole blood culture. The Kappa coefficients of agreement were 0.9819 (buffy coat PCR), 0.9458 (whole blood PCR) and 1.0 (buffy coat culture), which establishes their validity as alternative methods to routine blood culture in detecting bacteraemia. In addition, results showed that there was a direct correlation of WBC counts greater than 12,000 cells per mm3 to the occurrence of bacteraemia as detected by the four methods (p < 0.05).

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Digitial Object Identifier (DOI)

10.1590/s0074-02762010000200001

Disciplines

Biology

Keywords

Bacteremia; Blood—Examination; Leucocytes; Polymerase chain reaction

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