A total of 402 respiratory samples were tested by PCR and culture in this study. Respiratory samples were collected from CF patients attending the Tampa General Hospital (TGH; Florida, USA) and the Children's Healthcare of Atlanta (CHOA; Georgia, USA). At TGH samples were cultured on Burkholderia cepacia selective agar (Hardy Diagnostics, Santa Maria, CA), TSA with 5% Sheep Blood (aerobic and anaerobic conditions), MacConkey II Agar and Chocolate II Agar (BD Diagnostic Systems, Sparks, MD). The media used at CHOA was purchased from Remel (Lenexa, KS) and included Chocolate Agar (CHOC), TSA with 5% Sheep Blood (BAP), MacConkey Agar, Colistin Nalidixic Acid Agar, Mannitol Salt Agar and Burkholderia cepacia selective Agar. The CHOC and BAP were incubated in CO2, while the remaining media were incubated in an ambient air incubator. All plates were incubated at 35 °C. Cultured organisms were identified by the VITEK® MS system (BioMérieux, France). Semi-quantitative culture results were reported accordingly to the number of colonies of each species identified, such as “rare”, “light”, “moderate”, “many” or “heavy”. Respiratory residual specimens were stored at − 80 °C until they were tested by the new CF BDM Test.
Burkholderia cepacia selective agar
Manufactured by Hardy Diagnostics
Burkholderia cepacia selective agar is a microbiological culture medium used for the isolation and identification of Burkholderia cepacia, a bacterial species. It contains selective agents that inhibit the growth of unwanted microorganisms, allowing the targeted species to grow and be identified.
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2 protocols using burkholderia cepacia selective agar
1
Respiratory Pathogen Detection in CF Patients
2
Rapid Pneumonia Pathogen Profiling
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This study was approved by the Institutional Review Board and Ethics Committee of Human Microbiology Institute (Number: 018-4-AF). BAL or sputum biospecimens (1 mL) from patients with ventilator-associated, community-acquired pneumonia, cystic fibrosis, or chronic obstructive pulmonary disease were mixed with 1 mL of sterile H2O until homogeneity with a plastic swab. After homogenization, 25 µL of the suspension was directly plated onto TGV agar in each well of a 12-well plate and incubated at 37 °C, for 4, 18, and 24 h. For the control arm, 100 µL suspension were plated on 90 mm Petri dish with LB agar, Burkholderia cepacia-selective agar (Hardy Diagnostics), chocolate agar (Oxoid) or CHROMagar Staph aureus Medium (Becton, Dickinson and Company) and cultured according to laboratory recommendations at 37 °C for 24–72 h [8 (link)]. The results of the study were not reported to the physician, and no medical decisions were made on the basis of these data.
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