Comparison of culture and qPCR for the detection of Pseudomonas aeruginosa in not chronically infected cystic fibrosis patients

Deschaght, Pieter; Schelstraete, Petra; Santiago, Guido Lopes dos Santos; Simaey, Leen Van; Haerynck, Filomeen; Daele, S. Van; Wachter, Elke De; Malfroot, Anne; Lebecque, Patrick; Knoop, Christiane; Casimir, Georges; Boboli, Hedwige; Pierart, F.; Desager, Kristine; Vaneechoutte, Mario; Baets, Frans De

doi:10.1186/1471-2180-10-245

Cited by 25 publications

(27 citation statements)

References 32 publications

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“…Regarding the sample type, i.e. sputum, cough swab, throat swab or nasopharyngeal aspirate, we found no significant linkage between the sample type and dissimilarity between qPCR and culture sensitivity [26], while Logan et al [24] and McCulloch et al [27] found proportionally more qPCR positive/culture negative sputum samples (resp. 6.1% and 23%) in comparison with qPCR positive/culture negative throat swabs and cough swabs (resp.…”

Section: Sensitivity Of the Qpcr Formatsmentioning

confidence: 87%

PCR and the detection of Pseudomonas aeruginosa in respiratory samples of CF patients. A literature review

Deschaght¹,

Daele

Baets

et al. 2011

Journal of Cystic Fibrosis

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Previous studies proved the importance of rapid antibacterial intervention in case of Pseudomonas aeruginosa detection in respiratory samples of cystic fibrosis patients. To improve the early detection of P. aeruginosa, several culture, PCR and serology based approaches have been compared. Because an increasing number of routine microbiology laboratories have access to real-time PCR (qPCR), we reviewed the specificity and sensitivity of published PCR formats. The importance of choice of DNA-extraction methods and PCR formats and of the validation of their specificity and sensitivity with clinical samples is stressed.

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Section: Sensitivity Of the Qpcr Formatsmentioning

confidence: 87%

PCR and the detection of Pseudomonas aeruginosa in respiratory samples of CF patients. A literature review

Deschaght¹,

Daele

Baets

et al. 2011

Journal of Cystic Fibrosis

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“…2007/503). Study was performed in accordance with approved national and international guidelines; written informed consent was obtained from all the patients > 18 years or from the parents and children older than 12 years, as previously reported [16,17]. …”

Section: Methodsmentioning

confidence: 99%

“…aeruginosa [1], and received the same antimicrobial treatment: Tazocin ® (Piperacillin/Tazobactam 4000mg/500mg) by intravenous administration during 15 days. Sputum samples were collected as previously reported [16,17]. …”

Section: Methodsmentioning

confidence: 99%

Effects of Propidium Monoazide (PMA) Treatment on Mycobiome and Bacteriome Analysis of Cystic Fibrosis Airways during Exacerbation

et al. 2016

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Introduction and PurposePropidium monoazide (PMA)-pretreatment has increasingly been applied to remove the bias from dead or damaged cell artefacts, which could impact the microbiota analysis by high-throughput sequencing. Our study aimed to determine whether a PMA-pretreatment coupled with high-throughput sequencing analysis provides a different picture of the airway mycobiome and bacteriome.Results and DiscussionWe compared deep-sequencing data of mycobiota and microbiota of 15 sputum samples from 5 cystic fibrosis (CF) patients with and without prior PMA-treatment of the DNA-extracts. PMA-pretreatment had no significant effect on the entire and abundant bacterial community (genera expressed as operational taxonomic units (OTUs) with a relative abundance greater than or equal to 1%), but caused a significant difference in the intermediate community (less than 1%) when analyzing the alpha biodiversity Simpson index (p = 0.03). Regarding PMA impact on the airway mycobiota evaluated for the first time here; no significant differences in alpha diversity indexes between PMA-treated and untreated samples were observed. Regarding beta diversity analysis, the intermediate communities also differed more dramatically than the total and abundant ones when studying both mycobiome and bacteriome. Our results showed that only the intermediate (or low abundance) population diversity is impacted by PMA-treatment, and therefore that abundant taxa are mostly viable during acute exacerbation in CF. Given such a cumbersome protocol (PMA-pretreatment coupled with high-throughput sequencing), we discuss its potential interest within the follow-up of CF patients. Further studies using PMA-pretreatment are warranted to improve our “omic” knowledge of the CF airways.

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“…Our study also indicates the high potential of SF in the diagnosis of candidemia -we observed a signifi cantly higher detection rate of SF for C. albicans. [16,17,20,28,29]. A possible explanation for higher detection rate of P. aeruginosa is that in cases of bacteraemia, it is almost always isolated from the aerobic blood culture bottles only, thereby signifi cantly decreasing the likelihood of detection [30].…”

Section: Discussionmentioning

confidence: 99%

Diagnostic performance and therapeutic impact of LightCycler SeptiFast assay in patients with suspected sepsis

Herne¹,

Nelovkov²,

Kutt³

et al. 2013

European Journal of Microbiology and Immunology

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Rapid and reliable identification of pathogens is very important in the management of septic patients. We retrospectively evaluated the diagnostic accuracy and clinical utility of a multiplex real-time polymerase chain reaction (PCR) assay (SeptiFast (SF)) in patients with suspected sepsis in a tertiary care hospital in Tallinn, Estonia. A total of 160 blood samples from 144 patients were included in the study. SF results were compared with corresponding blood culture (BC) results. The concordance between SF and BC was 78.8%. The rate of positive results was significantly higher in SF than in BC (33.7% vs. 21.2%, respectively; p < 0.001). A total of 27 samples were found positive by both SF and BC, 27 by SF only, and seven by BC only. Of a total of 83 microorganisms detected SF identified 71, and BC 42 (p < 0.001). SF detected markedly more patients with candidemia: 11 patients were detected by SF compared to four patients by BC. Antimicrobial treatment was changed in 21 (38.9%) of 54 SF positive cases. In conclusion, our results demonstrated the high diagnostic accuracy of SF in detection of sepsis pathogens. In conjunction with its impact on therapeutic decisions, SF proved to be a useful adjunct to conventional blood culture in the diagnosis of sepsis etiology.

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Comparison of culture and qPCR for the detection of Pseudomonas aeruginosa in not chronically infected cystic fibrosis patients

Cited by 25 publications

References 32 publications

PCR and the detection of Pseudomonas aeruginosa in respiratory samples of CF patients. A literature review

PCR and the detection of Pseudomonas aeruginosa in respiratory samples of CF patients. A literature review

Effects of Propidium Monoazide (PMA) Treatment on Mycobiome and Bacteriome Analysis of Cystic Fibrosis Airways during Exacerbation

Diagnostic performance and therapeutic impact of LightCycler SeptiFast assay in patients with suspected sepsis

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