Rapid detection of pathogens in blood culture bottles by real-time PCR in conjunction with the pre-analytic tool MolYsis

Rapid Detection of bla KPC Carbapenemase Genes by Internally Controlled Real-Time PCR Assay Using Bactec Blood Culture Bottles

“…The assay, which includes human gene RNase P as an internal control, has been optimized to circumvent the inhibitors present in the broth of Bactec bottles and to be completed in less than 4 h. This assay also showed that once BSA fraction V (96%) was utilized as an inhibitor reliever, no manipulation of the samples was required before or after manual or automated extraction, as previously reported (10,14).…”

“…The assay is sensitive for concentrations 1 log unit below the limit of detection of the Bactec system (at least 10 4 CFU/ml); thus, it will detect the bla KPC -positive bacteria if they are present in the bottles (26). The analytical sensitivity of the multiplex bla KPC /RNase P assay was comparable to sensitivities achieved for detection of 10 CFU/reaction mixture of both E. coli and Klebsiella pneumoniae in blood culture bottles (10,17). However; the earlier studies did not determine the analytical sensitivity in human blood-spiked Bactec bottles.…”

“…With the advent of qPCR, the time to bacterial identification and detection of drug resistance has been reduced to 4 to 6 h after a positive blood culture has turned positive (22,24,26,28). However, the presence of PCR inhibitors in the blood culture bottles has thus far reduced the sensitivity of PCR assays (10). Rapid detection of bacterial resistance mechanisms in blood culture bottles can assist the physician in both patient management and infection control.…”

Improved Sensitivity for Molecular Detection of Bacterial and Candida Infections in Blood

“…This can be very challenging if genomic DNA is copurified with targeted microbial DNA (11)(12)(13). A number of solutions to this problem have been proposed (14)(15)(16)(17)(18)(19) but, to date, none have achieved the desired sensitivity (20). It should be noted that many of these methods do appear to capture a significantly higher number of total positives than culture, and the "extra" detections are often correlated with clinical indications of bloodstream infection; however, an inability to capture all positives detected by culture (false negatives) has been an issue.…”

Application of multiplex real-time PCR and Fluorescence Resonance Energy Transfer for the detection and differentiation of the most frequent causative agents of systemic infections from biological fluids

“…The test performs more than one PCR for the correct detection of the bacteria moreover, the fungal identification is requires a parallel PCR. This fact increases the time of the identification [88], [89].…”