title: Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease.
abstract: BACKGROUND
  Nested PCR can be used to determine the status of Pneumocystis jirovecii infection in other lung diseases.
  
  
  OBJECTIVE
  This study sought to detect a target DNA fragment (mitochondrial large subunit rRNA or mtL SUrRNA) of P. jirovecii in patients with lung disease who underwent bronchoscopy with collection of bronchoalveolar lavage (BAL).
  
  
  METHODS
  The results from toluidine blue staining were compared with those obtained using molecular methods that included an "in house" DNA extraction procedure, PCR and nested PCR.
  
  
  RESULTS
  Fifty-five BAL samples from patients with atypical chest X-rays were screened for P. jirovecii. None of the samples was positive for P. jirovecii using toluidine blue staining. In contrast, P. jirovecii DNA was detected by nested PCR in BAL samples from 36 of 55 patients (65.5%). The lung diseases in the patients included cancer, pneumonia, tuberculosis, and chronic obstructive pulmonary disease (COPD). Other chronic problems in the patients included hypertension, diabetes, smoking, and alcoholism.
  
  
  CONCLUSIONS
  Nested PCR showed high sensitivity for detecting P. jirovecii, especially when compared with toluidine blue staining. Using this method, P. jirovecii infection was detected in HIV-negative patients with lung disease.
authors.affiliation: Laboratório de Pesquisa em AIDS, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil. Electronic address: stin.santos@gmail.com.
authors.forename: Cristina Rodrigues
authors.initials: CR
authors.lastname: Santos
authors.affiliation: Laboratório de Pesquisa em AIDS, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Ângela M
authors.initials: ÂM
authors.lastname: de Assis
authors.affiliation: Laboratório de Patologia Clínica/Micologia and Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Edson A
authors.initials: EA
authors.lastname: Luz
authors.affiliation: Laboratório de Patologia Clínica/Micologia and Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Luzia
authors.initials: L
authors.lastname: Lyra
authors.affiliation: Serviço de Broncoscopia, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Ivan F
authors.initials: IF
authors.lastname: Toro
authors.affiliation: Serviço de Broncoscopia, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: José Claudio C
authors.initials: JCC
authors.lastname: Seabra
authors.affiliation: Serviço de Broncoscopia, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Dira H
authors.initials: DH
authors.lastname: Daldin
authors.affiliation: Serviço de Broncoscopia, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Tathiane U
authors.initials: TU
authors.lastname: Marcalto
authors.affiliation: Serviço de Broncoscopia, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Marcos T
authors.initials: MT
authors.lastname: Galasso
authors.affiliation: Serviço de Broncoscopia, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Ronaldo F
authors.initials: RF
authors.lastname: Macedo
authors.affiliation: Laboratório de Patologia Clínica/Micologia and Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Angélica Z
authors.initials: AZ
authors.lastname: Schreiber
authors.affiliation: Laboratório de Pesquisa em AIDS, Hospital de Clínicas, Universidade Estadual de Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil.
authors.forename: Francisco H
authors.initials: FH
authors.lastname: Aoki
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000437:Alcoholism / Q000453:epidemiology; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D001999:Bronchoscopy; D004396:Coloring Agents; D015897:Comorbidity; D004271:DNA, Fungal / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis; D003920:Diabetes Mellitus / Q000453:epidemiology; D005260:Female; D018023:HIV Seronegativity; D006801:Humans; D006973:Hypertension / Q000453:epidemiology; D008171:Lung Diseases / Q000453:epidemiology*; D008297:Male; D008875:Middle Aged; D045363:Pneumocystis carinii / Q000235:genetics / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000453:epidemiology / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012907:Smoking / Q000453:epidemiology; D013194:Staining and Labeling; D014048:Tolonium Chloride; D055815:Young Adult
chemical_list: D004396:Coloring Agents; D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D014048:Tolonium Chloride
keywords: BAL; LBA; Nested PCR; PCR anidada; Pneumocystis jirovecii