Pseudomonas aeruginosa and Burkholderia cepacia cannot be detected by PCR in the breath condensate of patients with cystic fibrosis.

The collection of sputum for microbiological examination in young cystic fibrosis patients can be very difficult. However, a knowledge of bacterial flora colonizing the patient's airways is of paramount importance for proper antimicrobial therapy. It is also known that cystic fibrosis patients colonized by Pseudomonas species have a poorer prognosis than Pseudomonas-negative patients. Noninvasive ways of diagnosing airway inflammation that require only minimal cooperation of the patient might yield new possibilities for early detection of airway colonisation. The breath condensate method as a noninvasive diagnostic technique seems especially appropriate for use in children. Therefore, the aim of this study was to evaluate whether the breath condensate method could be used for detection of Pseudomonas species in children with cystic fibrosis. In total, 32 breath condensate and seven sputum samples were obtained from 13 cystic fibrosis patients with Pseudomonas aeruginosa- or Burkholderia cepacia-positive sputum culture (20 samples were obtained during forced expiration). PCR for combined detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Burkholderia cepacia was performed. PCR results of all breath condensate samples were negative for Pseudomonas aeruginosa, Stenotrophomonas maltophilia, or Burkholderia cepacia, while all sputum sample results were positive. A minimum DNA quantity of 10 fg could be detected in dilution series of the positive control group. We conclude that the breath condensate method cannot be used as a tool for detection of Pseudomonas species.

Capofungin therapy for Aspergillus lung infection in a boy with chronic granulomatous disease.

Neutrophils of patients with chronic granulomatous disease (CGD) are profoundly defective in killing microorganisms. As in patients with chemotherapy-induced neutropenia, CGD patients are highly susceptible to life-threatening invasive Aspergillus infections, and conventional antimycotic treatment is not always successful. The new drug, caspofungin, was used to treat Aspergillus lung infection in one child with CGD.

[Molecular biological identification of Cuninghamella spec]. / Molekularbiologische Identifizierung von Cunninghamella spec.

Among the genus Cunninghamella, so far C. bertholletiae is known to be the only clinically relevant species. Correct identification of C. bertholletiae is not possible with classical methods. PCR and sequencing of the internal transcribed spacer (ITS) region was used to identify seven of nine clinical isolates as C. bertholletiae and two as C. echinulata. Also an isolate of the surrounding area of one patient infected with C. echinulata could be identified as C. echinulata. High homology in the ITS region was found within the isolates of C. bertholletiae. Within the species C. echinulata and C. elegans a differentiation on subspecies level was achieved by an analysis of restriction fragment length polymorphism of the ITS amplicons after incubation with TaqI and HinfI. Similar results were obtained by PCR fingerprinting of the complete DNA with the single microsatellite DNA primers (GTG)5 and (GAC)5. For the first time C. echinulata could be identified as agent of zygomycosis in humans.