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Titolo:
RAPID DETECTION OF PNEUMOCYSTIS-CARINII IN BRONCHOALVEOLAR LAVAGE SPECIMENS FROM HUMAN IMMUNODEFICIENCY VIRUS-INFECTED PATIENTS - USE OF A SIMPLE DNA EXTRACTION PROCEDURE AND NESTED PCR
Autore:
RABODONIRINA M; RAFFENOT D; COTTE L; BOIBIEUX A; MAYENCON M; BAYLE G; PERSAT F; RABATEL F; TREPO C; PEYRAMOND D; PIENS MA;
Indirizzi:
UNIV LYON 1,DEPT PARASITOL & MYCOL,8 AVE ROCKEFELLER F-69373 LYON 08 FRANCE HOP HOTEL DIEU,SERV HEPATOGASTROENTEROL & SIDOL F-69288 LYON FRANCE HOP HOTEL DIEU,SERV RADIOL F-69288 LYON FRANCE HOP CROIX ROUSSE,SERV MALAD INFECT & TROP F-69317 LYON FRANCE
Titolo Testata:
Journal of clinical microbiology
fascicolo: 11, volume: 35, anno: 1997,
pagine: 2748 - 2751
SICI:
0095-1137(1997)35:11<2748:RDOPIB>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE CHAIN-REACTION; INDUCED SPUTUM; AEROSOLIZED PENTAMIDINE; PNEUMONIA; DIAGNOSIS; AMPLIFICATION; IMMUNOFLUORESCENCE; SAMPLES; CHEMOPROPHYLAXIS; PERSISTENCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
40
Recensione:
Indirizzi per estratti:
Citazione:
M. Rabodonirina et al., "RAPID DETECTION OF PNEUMOCYSTIS-CARINII IN BRONCHOALVEOLAR LAVAGE SPECIMENS FROM HUMAN IMMUNODEFICIENCY VIRUS-INFECTED PATIENTS - USE OF A SIMPLE DNA EXTRACTION PROCEDURE AND NESTED PCR", Journal of clinical microbiology, 35(11), 1997, pp. 2748-2751

Abstract

We report on the development of a rapid nested PCR protocol far the detection of Pneumocystis carinii DNA in bronchoalveolar lavage (BAL) specimens in which the protocol included the use of a commercially available DNA extraction kit (GeneReleaser). GeneReleaser enabled us to obtain amplification-ready DNA within 20 min without requiring the purification of the DNA. The nested PCR was performed with the primers pAZ102-E, pAZ102-H, and pAZ102-L2 (A. E. Wakefield, F. J. Pixley, S. Banerji, K. Sinclair, R. F. Miller, E. R. Moxon, and J. M. Hopkin, Lancet 336:451-453, 1990.). Results were obtained in about 4 h with the adoption of denaturation, annealing, and extension steps shortened to 20 seconds. The sensitivity of the nested PCR was rested with a P. carinii cyst suspension and was found to he less than one cyst (one to eight nuclei). The detection limit was the same with the use of GeneReleaser or proteinase K-phenol chloroform for DNA extraction, The nested PCR assay was prospectively compared with staining with Giemsa and methenamine silver stains for the detection of P. carinii in 127 BAL samples from 105 human immunodeficiency virus-infected patients investigated foracute respiratory illness, Twenty-five BAL specimens (20%) were positive by staining and the nested PCR and 25 (20%) were negative by staining and positive by the nested PCR. These 25 BAL specimens with conflicting results were obtained from 23 patients, 82% of whom were receiving prophylactic therapy against P. carinii pneumonia (PCP). Only two patients were diagnosed with possible PCP. The final diagnosis was not PCP for 20 patients who were considered to be colonized or to have a low level of infection, This colonization is not of clinical importancebare is of epidemiological importance. Our rapid, simple, and sensitive amplification protocol may he performed in clinical laboratories for the routine diagnosis of PCP with BAL specimens.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 22/09/20 alle ore 14:24:36