RT Journal Article T1 Multicentre study highlighting clinical relevance of new high-throughput methodologies in molecular epidemiology of Pneumocystis jirovecii pneumonia A1 Esteves, F. A1 de Sousa, B. A1 Calderon, E. J. A1 Huang, L. A1 Badura, R. A1 Maltez, F. A1 Bassat, Q. A1 de Armas, Y. A1 Antunes, F. A1 Matos, O. K1 DNA pools K1 high-throughput molecular epidemiology K1 multilocus genotyping survey K1 Pneumocystis jirovecii K1 Dihydropteroate synthase gene K1 Dihydrofolate-reductase gene K1 F-sp hominis K1 Carinii-pneumonia K1 Cuban infants K1 South-africa AB Pneumocystis jirovecii causes severe interstitial pneumonia (PcP) in immunosuppressed patients. This multicentre study assessed the distribution frequencies of epidemiologically relevant genetic markers of P. jirovecii in different geographic populations from Portugal, the USA, Spain, Cuba and Mozambique, and the relationship between the molecular data and the geographical and clinical information, based on a multifactorial approach. The high-throughput typing strategy for P. jirovecii characterization consisted of DNA pooling using quantitative real-time PCR followed by multiplex-PCR/single base extension. The frequencies of relevant P. jirovecii single nucleotide polymorphisms (mt85, SOD110, SOD215, DHFR312, DHPS165 and DHPS171) encoded at four loci were estimated in ten DNA pooled samples representing a total of 182 individual samples. Putative multilocus genotypes of P. jirovecii were shown to be clustered due to geographic differences but were also dependent on clinical characteristics of the populations studied. The haplotype DHFR312T/SOD110C/SOD215T was associated with severe AIDS-related PcP and high P. jirovecii burdens. The frequencies of this genetic variant of P. jirovecii were significantly higher in patients with AIDS-related PcP from Portugal and the USA than in the colonized patients from Portugal, and Spain, and children infected with P. jirovecii from Cuba or Mozambique, highlighting the importance of this haplotype, apparently associated with the severity of the disease and specific clinical groups. Patients from the USA and Mozambique showed higher rates of DHPS mutants, which may suggest the circulation of P. jirovecii organisms potentially related with trimethoprim-sulfamethoxazole resistance in those geographical regions. This report assessed the worldwide distribution of P. jirovecii haplotypes and their epidemiological impact in distinct geographic and clinical populations. (C) 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved. PB Elsevier sci ltd SN 1198-743X YR 2016 FD 2016-03-25 LK http://hdl.handle.net/10668/18748 UL http://hdl.handle.net/10668/18748 LA en NO Esteves F, de Sousa B, Calderón EJ, Huang L, Badura R, Maltez F, et al. Multicentre study highlighting clinical relevance of new high-throughput methodologies in molecular epidemiology of Pneumocystis jirovecii pneumonia. Clin Microbiol Infect. 2016 Jun;22(6):566.e9-566.e19. DS RISalud RD Apr 15, 2025