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3 result(s) for "broad range PCR assay"
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Diagnostic performance of broad-range PCR in bacterial peritonitis
ObjectiveBacterial peritonitis (BP) is a serious complication commonly associated with cirrhosis and ascites, often leading to high mortality rates. Although these effects could be reduced with timely and appropriate antibiotics, traditional BP diagnosis relies on culture, often delaying targeted treatment. Therefore, the use of fast molecular assays holds the potential to enhance laboratory diagnosis. In this study, we assessed the diagnostic performaance of Molecular Culture ID, a broad PCR-based assay targeting the 16S-23S interspace rDNA region in the scope of BP diagnosis.MethodsThe residual material from 247 peritoneal fluid samples submitted for routine diagnostics was analyzed using Molecular Culture ID and compared alongside the standard of care (SOC) results.ResultsSample positivity and species identification outcomes of Molecular Culture ID were compared to those of SOC. Molecular Culture ID yielded 1.6x more positive samples than SOC. Percent positive agreement (PPA) between Molecular Culture ID and SOC at the sample level was 90.1% (IC 95%, 81.0% to 95.1%), and negative percent agreement (NPA) was 70.5% (IC 95%, 63.3% to 76.7%). At the species level, the PPA was 75.2% (95% CI 67.2% to 81.8%). Molecular Culture ID yielded 289 extra bacterial identifications, mainly anaerobic species. High leukocyte counts, indicative of infection, were concordant with Molecular Culture ID positivity.ConclusionMolecular Culture ID demonstrated enhanced BP diagnostic capabilities compared to SOC, with higher positivity rates, more comprehensive species identification for difficult to culture species and a high correlation with leukocyte counts.
Identification of population of bacteria from culture negative surgical site infection patients using molecular tool
Background Managing surgical site infections, with negative culture report in routine diagnosis is a common dilemma in microbiology accounting more than 30% worldwide. The present study attempted to identify the presence of bacterial spp. if any in wound aspirates/swabs of culture negative surgical site infections of hospitalised patients using molecular tools. Methods Ninety-seven patients with post-operative SSI whose wound swabs/aspirate were negative in the conventional aerobic culture after 72 h of incubation were analysed by 16S rRNA gene specific broad range PCR. The amplified DNA fragments were sequenced by Sanger DNA sequencing method and homology of the sequence were matched using NCBI BLAST (NCBI, USA) Results Of the 97 patients, 16S rRNA based broad range PCR assay could identify the presence of bacterial pathogen in 53(54.63%) cases, of which 29 isolates were supposed to be of viable but non-culturable bacteria (VBNC), 07 were of obligatory anaerobes and 13 were of unculturable bacteria, 04 were with poly bacterial infections. Conclusions Our study highlights the usefulness of PCR assay in detecting the presence of any VBNC, anaerobes and unculturable bacteria in SSI patients regardless of how well the bacteria may or may not grow in culture. Measures should be taken to use anaerobic culture system and PCR diagnosis along with conventional culture to detect the VBNC and unculturable bacteria where Gram stain is positive for better patient care.