Journal
OPEN FORUM INFECTIOUS DISEASES
Volume 2, Issue 3, Pages -Publisher
OXFORD UNIV PRESS INC
DOI: 10.1093/ofid/ofv080
Keywords
microsatellite; molecular epidemiology; pneumocystis; real-time PCR
Categories
Funding
- National Center for Advancing Translational Science CTSA NIH/NCATS [UNC ULTR000083]
- National Institute of Allergy and Infectious Diseases (NIAID) [R01 AI089819]
- University of North Carolina at Chapel Hill Center for AIDS Research, a National Institutes of Health [P30 AI50410]
- National Heart, Lung, and Blood Institute (NHLBI) [R01 HL090335, HL090335-02S109]
- National Institute of General Medicine Sciences [T32 GM008719, T32 GM007092]
- IDSA Medical Scholars Program
- NHLBI [R01 HL090335, K24 HL087713, U01 HL098964]
- NIAID [K08 AI100924]
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Pneumocystis jirovecii is a symbiotic respiratory fungus that presents in 2 clinical forms: pneumonia in immunocompromised patients or colonization, defined by the presence of the organism without associated clinical symptoms. Currently, diagnosis requires invasive bronchoscopy, which may not be available in some settings and is inappropriate for detecting colonization in healthy individuals. Noninvasive diagnostic techniques and molecular strain typing tools that can be used on these samples are critical for conducting studies to better understand transmission. We evaluated 2 real-time polymerase chain reaction (PCR) assays targeting dihydropteroate synthase and the major surface glycoprotein for detection in 77 oropharyngeal washes (OPWs) from 43 symptomatic human immunodeficiency virus-infected patients who underwent bronchoscopy. We also evaluated the ability of a new microsatellite (MS) genotyping panel to strain type infections from these samples. Each PCR used individually provided a high sensitivity (> 80%) for detection of pneumonia but a modest specificity (< 70%). When used in combination, specificity was increased to 100% with a drop in sensitivity (74%). Concentration of organisms by PCR in the OPWtended to be lower in colonized individuals compared with those with pneumonia, but differences in concentration could not clearly define colonization in symptomatic individuals. Oropharyngeal wash samples were genotyped using 6 MSs with >= 4 alleles successfully genotyped in the majority of colonized patients and >= 5 alleles in patients with pneumonia. The MS profile was consistent over time within patients with serial OPWs analyzed. Microsatellite genotyping on noninvasive samples may aid in studying the molecular epidemiology of this pathogen without requiring invasive diagnostic techniques.
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