title: Oropharyngeal samples for detection of Pneumocystis carinii by DNA amplification.
abstract: Pneumocystis carinii pneumonia is a major complication of T-lymphocyte immune deficiency. Restriction of the disease to the alveolar spaces and failure to culture R. carinii has hindered simple diagnostic methods. We have developed a specific DNA amplification method for P. carinii and shown diagnostic sensitivity and specificity exceeding 95% for pneumocystis pneumonia when applied to bronchoscopic lavage and hypertonic saline induced sputum. We here report application of DNA amplification to simple oropharyngeal samples in 31 HIV-positive patients with respiratory illness. P. carinii-specific DNA was detected in 10 of 18 (56%) patients with pneumocystis pneumonia by ethidium bromide stained gels and 14 of 18 (78%) patients by the more sensitive technique of oligoblotting. P. carinii DNA was not detected in samples from 13 patients with other respiratory diagnoses. An oropharyngeal sample offers a simple specimen for detecting P. carinii by DNA amplification; refinements of technique and calibration may allow its development for accurate diagnostic and epidemiological work.
authors.affiliation: Department of Paediatrics, John Radcliffe Hospital, Oxford.
authors.forename: A E
authors.initials: AE
authors.lastname: Wakefield
authors.forename: R F
authors.initials: RF
authors.lastname: Miller
authors.forename: L A
authors.initials: LA
authors.lastname: Guiver
authors.forename: J M
authors.initials: JM
authors.lastname: Hopkin
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis* / Q000188:drug therapy; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D005784:Gene Amplification; D006679:HIV Seropositivity; D015497:HIV-1; D006801:Humans; D009960:Oropharynx / Q000382:microbiology*; D011010:Pneumocystis / Q000235:genetics / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis* / Q000188:drug therapy; D012680:Sensitivity and Specificity