Detection of Pneumocystis carinii by DNA amplification in human immunodeficiency virus-positive patients.
The opportunistic pathogen Pneumocystis carinii (PC) is a frequent cause of a life-threatening pneumonia in human immunodeficiency virus (HIV)-infected individuals and in other immunocompromised hosts. Specimens obtained from 128 bronchoalveolar lavage (BAL) fluid samples from 123 HIV-positive patients with pulmonary disease and undergoing a diagnostic bronchoscopy were evaluated to detect this organism. We have developed a rapid DNA extraction procedure for nested polymerase chain reaction (PCR) using two sets of primers (pAZ102-E, pAZ102-H and P1 = 5'-CTAGGATATAGCTGGTTTTC-3' and P2 = 5'-TCGACTATCTAGCTTATCGC-3'). The results were compared using cytological techniques (direct wet mount, Giemsa, toluidine blue O) and related to the clinical follow-up of patients. The nested PCR had a 91% sensitivity and a 93% specificity. The effect of chemoprophylaxis and the evaluation of the follow-up of patients are discussed. Nested PCR may represent an important additional tool, along with current cytological methods, for the detection of P. carinii; however, at present it cannot replace routine microbiological methods more simple and less expensive.
Savoia
D
D
Department of Clinical and Biological Sciences, University of Torino, Italy.
Millesimo
M
M
Cassetta
I
I
Forno
B
B
Caramello
P
P
D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis / Q000821:virology*; D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000821:virology*; D001999:Bronchoscopy; D004279:DNA, Viral / Q000032:analysis*; D006679:HIV Seropositivity; D006801:Humans; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004279:DNA, Viral