Pulmonary colonisation with Pneumocystis carinii in an immunosuppressed HIV-negative patient: detection and typing of the fungus by PCR

Mostly Pneumocystis carinii isolates from patients with acute pneumocystosis (PCP) have been typed until now. This report describes the typing of P. carinii organisms obtained from an HIV-negative patient without PCP. The patient underwent a broncho-alveolar lavage (BAL) to investigate an abnormal chest X-ray. He was diagnosed with sarcoidosis. However, a low level of P. carinii organisms undetectable by microscopy was detected in BAL fluid by two subsequent nested PCR assays: one assay amplifying a portion of the mitochondrial large subunit RNA gene and a second one amplifying the internal transcribed spacers (ITS) 1 and ITS 2 of the nuclear rRNA operon. This low level of the fungus did not reflect acute PCP. Indeed, the clinical outcome was improvement despite the absence of specific treatment. The patient was considered to be only colonised by the fungus. Analysis of sequences of ITS PCR products led to identification of genotype Gg. This information constitutes the first data concerning P. carinii ITS genotype from a patient without acute PCP and HIV. This type has been described previously in AIDS patients diagnosed with PCP. These results show that PCR and ITS genotyping could represent efficient tools for the further investigation of the role played by HIV-negative patients with pulmonary colonisation in the human reservoir of P. carinii.