Evaluation of the xpert carba-R (cepheid) assay using contrived bronchial specimens from patients with suspicion of ventilator-associated pneumonia for the detection of prevalent carbapenemases

Abstract

There is a critical need for rapid diagnostic methods for multidrug-resistant (MDR) pathogens in patients with a suspicion of ventilator-associated pneumonia (VAP). The Xpert Carba-R detects 5 targets for carbapenemase-producing organisms (blaKPC , blaNDM , blaVIM , blaOXA-48 , and blaIMP-1 ). Our objective was to evaluate the performance of this assay directly on bronchial aspirates and to correlate the cycle number for a positive result (Ct) with the bacterial count. Bronchial aspirates from patients with a suspicion of VAP were spiked with a dilution of 1 of 4 MDR organisms carrying the resistance genes detected by the test prepared to a final concentration of 102 -105 cfu/mL. We used a ROC curve and provided areas under the curve (AUC) with their 95% confidence intervals (CI). A point of maximum sensitivity (Se) and specificity (Sp) was derived and validity indices were calculated. One hundred contrived tests were performed. Se and Sp were 100% for all bacterial counts. A positive sample with a Ct ≤24.7 corresponded to a count ≥105 cfu/mL; if the Ct was within the range >24.7-≤26.9, this corresponded to a count ≥104 cfu/mL. When the Ct was >26.9, this corresponded to a count <104 cfu/mL. The Xpert Carba-R detects carbapenemase-producing organisms directly in contrived bronchial aspirates. Still, an important issue to consider is that the number of gene copies may vary according to many factors in vivo. If confirmed in further studies, the strong correlation observed between Ct values and the results of semiquantitative cultures suggests this test could serve to differentiate between infection and colonization in routine clinical practice.