ArticleBMC infectious diseases2018
Detection of viable but non-culturable Pseudomonas aeruginosa in cystic fibrosis by qPCR: a validation study.
Article in BMC infectious diseases, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 41 citations in OpenAlex.
- Evidence of cefiderocol-persistence inBiofilm · 2026Article
- Bioelectrochemical detection of viable but non-culturable bacterial cells on screen-printed carbon electrodes.Biofilm · 2026Article
- Molecular endotyping in people with bronchiectasis based on response to antibiotic treatment: iBEST study.ERJ open research · 2025Article
- Changing profile of bacterial infection and microbiome in cystic fibrosis: when to use antibiotics in the era of CFTR-modulator therapy.European respiratory review : an official journal of the European Respiratory Society · 2024Review
- Clinical relevance of Pseudomonas aeruginosa viable but non-culturable forms in cystic fibrosis.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2024Article
- Strategies to Overcome Antimicrobial Resistance in Nosocomial Infections, A Review and Update.Infectious disorders drug targets · 2024Review
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- Detection of respiratory pathogenic bacterial nucleic acid detection by Loop-mediated Isothermal Amplification in patients with bacterial pulmonary infections.Practical laboratory medicine · 2023Article
- Involvement of Acquired Tobramycin Resistance in the Shift to the Viable but Non-Culturable State inInternational journal of molecular sciences · 2023Article
- Accurate, Sensitive, and Rapid Detection of Pseudomonas aeruginosa Based on CRISPR/Cas12b with One Fluid-Handling Step.Microbiology spectrum · 2023Article
- Bacterial Culture Underestimates Lung Pathogen Detection and Infection Status in Cystic Fibrosis.Microbiology spectrum · 2022Article
- From genotype to phenotype: adaptations ofMicrobial genomics · 2021Review
- Contribution of Drugs Interfering with Protein and Cell Wall Synthesis to the Persistence ofInternational journal of molecular sciences · 2021Article
- A Rapid and Sensitive Detection Method forFrontiers in cellular and infection microbiology · 2021Article
- A Fluorinated Analogue of Marine Bisindole Alkaloid 2,2-Bis(6-bromo-1Pharmaceuticals (Basel, Switzerland) · 2020Article
- Role of Tobramycin in the Induction and Maintenance of Viable but Non-CulturableAntibiotics (Basel, Switzerland) · 2020Article
- Methods for Extraction and Detection of Pf Bacteriophage DNA from the Sputum of Patients with Cystic Fibrosis.PHAGE (New Rochelle, N.Y.) · 2020Article
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Authors and funding
9 authors at 4 institutions in 1 country.
Funding
Abstract
backgroundRoutine culture-based diagnosis of Pseudomonas aeruginosa lung infection in Cystic Fibrosis (CF) patients can be hampered by the phenotypic variability of the microorganism, including its transition to a Viable But Non-Culturable (VBNC) state. The aim of this study was to validate an ecfX-targeting qPCR protocol developed to detect all viable P. aeruginosa bacteria and to identify VBNC forms in CF sputum samples.
methodsThe study involved 115 P. aeruginosa strains of different origins and 10 non-P. aeruginosa strains and 88 CF sputum samples, 41 Culture-Positive (CP) and 47 Culture-Negative (CN). Spiking assays were performed using scalar dilutions of a mixture of live and dead P. aeruginosa ATCC 9027 and a pooled P. aeruginosa-free sputum batch. Total DNA from sputum samples was extracted by a commercial kit, whereas a crude extract was obtained from the broth cultures. Extracellular DNA (eDNA) interference was evaluated by comparing the qPCR counts obtained from DNase-treated and untreated aliquots of the same samples. The statistical significance of the results was assessed by the Wilcoxon test and Student's t test.
resultsThe newly-developed qPCR protocol identified 96.6% of the P. aeruginosa isolates; no amplification was obtained with strains belonging to different species. Spiking assays supported protocol reliability, since counts always matched the amount of live bacteria, thus excluding the interference of dead cells and eDNA. The protocol sensitivity threshold was 70 cells/ml of the original sample. Moreover, qPCR detected P. aeruginosa in 9/47 CN samples and showed higher bacterial counts compared with the culture method in 10/41 CP samples.
conclusionsOur findings demonstrate the reliability of the newly-developed qPCR protocol and further highlight the need for harnessing a non-culture approach to achieve an accurate microbiological diagnosis of P. aeruginosa CF lung infection and a greater understanding of its evolution.
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