32 HL07751 (B.A.M.), F32 A1801082 (B.A.M.), and R37 AI38296 (T.S.D.) as well as the Elizabeth B. Lamb Center for Pediatric Research. Added support was offered by the Vanderbilt Institute for Clinical and Translational Analysis (UL1 RR024975), the Vanderbilt-Ingram Cancer Center (CA68485), the Vanderbilt Flow Cytometry Shared Resource (DK058404), along with the Vanderbilt Cell Imaging Shared Resource (DK20593).
Multilocus Microsatellite Genotyping Array for Investigation of Genetic Epidemiology of Pneumocystis jiroveciiChristian M. Parobek,a,b Linda Y. Jiang,c Jaymin C. Patel,d Miriam J. Alvarez-Mart ez,e Jose M. Miro,f William Worodria,g Alfred Andama,g Serena Fong,h Laurence Huang,h,i Steven R. Meshnick,d Steve M. Taylor,d,j Jonathan J. JulianokSchool of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USAa; Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USAb; School of Medicine, Case Western University, Cleveland, Ohio, USAc; Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina, Chapel Hill, North Carolina, USAd; Hospital Cl ic-CRESIB (Barcelona Centre for International Overall health Analysis), Universitat de Barcelona, Barcelona, Spaine; Hospital Cl ic-IDIBAPS (Institut d’Investigaci?Biom ica August Pi i Sunyer), Universitat de Barcelona, Barcelona, Spainf; Makerere University College of Wellness Sciences and Mulago Hospital, Kampala, Ugandag; HIV/AIDS Division, San Francisco General Hospital, University of California, San Francisco, California, USAh; Division of Pulmonary and Important Care Medicinei and Division of Infectious Diseases and International Wellness,j Duke University Health-related Center, Durham, North Carolina, USA; Division of Infectious Illnesses, College of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USAkPneumocystis jirovecii is usually a symbiotic respiratory fungus that causes pneumonia (PcP) in immunosuppressed sufferers. Mainly because P. jirovecii cannot be reliably cultured in vitro, it has proven tough to study and gaps in our understanding on the organism persist. The release of a draft genome for the organism opens the door for the improvement of new genotyping approaches for studying its molecular epidemiology and worldwide population structure. We identified and validated eight putatively neutral microsatellite markers and 1 microsatellite marker linked for the dihydropteroate synthase gene (dhps), the enzymatic target of sulfa drugs applied for PcP prevention and therapy. Applying these tools, we analyzed P. jirovecii isolates from HIV-infected individuals from 3 geographically distant populations: Uganda, the Usa, and Spain. Amongst the eight neutral markers, we observed high levels of allelic heterozygosity (typical He, 0.Thalidomide 5-fluoride Chemscene 586 to 0.1175052-07-9 Chemscene 842).PMID:33742107 Constant with past reports, we observed restricted global population structuring, with only the Ugandan isolates displaying minor differentiation from the other two populations. In Ugandan isolates that harbored mutations in dhps, the microsatellite locus linked to dhps demonstrated a depressed He, consistent with positive directional selection for sulfa resistance mutations. Using a subset of these microsatellites, analyses of individual and paired samples from infections in San Francisco, CA, showed trustworthy typeability within a single infection and high discriminatory power in between infections. These characteristics recommend that this novel microsatellite ty.