Tags

Type your tag names separated by a space and hit enter

Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease.
Rev Iberoam Micol. 2017 Apr - Jun; 34(2):83-88.RI

Abstract

BACKGROUND

Nested PCR can be used to determine the status of Pneumocystis jirovecii infection in other lung diseases.

AIMS

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.

Links

Publisher Full Text

Authors+Show Affiliations

Santos CR
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.
de Assis ÂM
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.
Luz EA
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.
Lyra L
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.
Toro IF
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.
Seabra JCC
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.
Daldin DH
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.
Marcalto TU
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.
Galasso MT
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.
Macedo RF
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.
Schreiber AZ
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.
Aoki FH
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.

MeSH

AdultAgedAged, 80 and overAlcoholismBronchoalveolar Lavage FluidBronchoscopyColoring AgentsComorbidityDNA, FungalDNA, MitochondrialDiabetes MellitusFemaleHIV SeronegativityHumansHypertensionLung DiseasesMaleMiddle AgedPneumocystis cariniiPneumonia, PneumocystisPolymerase Chain ReactionSensitivity and SpecificitySmokingStaining and LabelingTolonium ChlorideYoung Adult

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28214275

Citation

Santos, Cristina Rodrigues, et al. "Detection of Pneumocystis Jirovecii By Nested PCR in HIV-negative Patients With Pulmonary Disease." Revista Iberoamericana De Micologia, vol. 34, no. 2, 2017, pp. 83-88.
Santos CR, de Assis ÂM, Luz EA, et al. Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease. Rev Iberoam Micol. 2017;34(2):83-88.
Santos, C. R., de Assis, Â. M., Luz, E. A., Lyra, L., Toro, I. F., Seabra, J. C. C., Daldin, D. H., Marcalto, T. U., Galasso, M. T., Macedo, R. F., Schreiber, A. Z., & Aoki, F. H. (2017). Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease. Revista Iberoamericana De Micologia, 34(2), 83-88. https://doi.org/10.1016/j.riam.2015.12.002
Santos CR, et al. Detection of Pneumocystis Jirovecii By Nested PCR in HIV-negative Patients With Pulmonary Disease. Rev Iberoam Micol. 2017 Apr - Jun;34(2):83-88. PubMed PMID: 28214275.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease. AU - Santos,Cristina Rodrigues, AU - de Assis,Ângela M, AU - Luz,Edson A, AU - Lyra,Luzia, AU - Toro,Ivan F, AU - Seabra,José Claudio C, AU - Daldin,Dira H, AU - Marcalto,Tathiane U, AU - Galasso,Marcos T, AU - Macedo,Ronaldo F, AU - Schreiber,Angélica Z, AU - Aoki,Francisco H, Y1 - 2017/02/15/ PY - 2014/11/16/received PY - 2015/11/08/revised PY - 2015/12/01/accepted PY - 2017/2/19/pubmed PY - 2018/4/24/medline PY - 2017/2/19/entrez KW - BAL KW - LBA KW - Nested PCR KW - PCR anidada KW - Pneumocystis jirovecii SP - 83 EP - 88 JF - Revista iberoamericana de micologia JO - Rev Iberoam Micol VL - 34 IS - 2 N2 - BACKGROUND: Nested PCR can be used to determine the status of Pneumocystis jirovecii infection in other lung diseases. AIMS: 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. SN - 2173-9188 UR - https://www.unboundmedicine.com/medline/citation/28214275/Detection_of_Pneumocystis_jirovecii_by_nested_PCR_in_HIV_negative_patients_with_pulmonary_disease_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1130-1406(16)30053-5 DB - PRIME DP - Unbound Medicine ER -