Blinded comparison of a direct immunofluorescent monoclonal antibody staining method and a Giemsa staining method for identification of Pneumocystis carinii in induced sputum and bronchoalveolar lavage specimens of patients infected with human immunodeficiency virus (original) (raw)
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Journal of clinical microbiology, 1995
The detection of Pneumocystis carinii DNA by PCR was compared with routine cytologic staining techniques (CYT). A total of 284 clinical respiratory specimens, including 137 bronchoalveolar lavage (BAL), 63 bronchoalveolar washing, 63 sputum, and 21 induced sputum samples, obtained from patients with or at high risk for human immunodeficiency virus infection were evaluated. Eighty specimens were positive by PCR, and 69 were positive by CYT. PCR was able to detect P. carinii in more bronchoalveolar washing specimens (15 versus 11) and in comparable BAL specimens (53 versus 54) compared with CYT. PCR was particularly more sensitive than CYT in detecting P. carinii in expectorated sputum (12 versus 4 samples). Of the 19 patients whose respiratory specimens were positive for P. carinii by PCR but negative by CYT, 5 had P. carinii pneumonia (PCP) confirmed by subsequent BAL and transbronchial or mediastinal lymph node biopsy and 9 had a clinical course highly suggestive of acute PCP. Elev...
Parasitology Research, 2004
The polymerase chain reaction (PCR) technique was compared with Wright-Giemsa (WG), Gomori methenamine silver (GMS) stains and an immunofluorescence assay (IFA) for detection of Pneumocystis carinii in immuno-compromised patients. Specimens of 21 bronchoalveolar lavages (BAL) and 139 sputum samples, were obtained from 157 patients (38 with AIDS and 119 with HIV) from four hospitals in Khon Kaen, Thailand. A true positive required at least two positives by techniques considered gold standard tests. Eleven (52.38%) BAL and 13 (9.35%) sputum specimens were positive. PCR produced the highest sensitivity and negative predictive values for the BAL (100% for each) vs. sputum samples at 84.62 and 98.41 percent, respectively. The specificity of PCR was 90% and 98.41% for BAL and sputum samples, respectively. We suggest PCR is an important tool for the epidemiological study of P. carinii in high-risk individuals.
Brazilian Journal of Infectious Diseases, 2007
Induced sputum is a useful technique for assessing airway inflammation, but its role in the diagnosis of lung disease in immunosuppressed patients needs further investigation. This study compared the use of induced sputum and BAL in the diagnosis of pneumocystosis, in HIV patients. From January 1, 2001, to December 30, 2002, HIV-positive patients older than 14 were evaluated at a hospital in Florianópolis, Santa Catarina, Brazil. Patients with respiratory symptoms for seven days or longer, with a normal or abnormal chest X-ray, and those without respiratory symptoms but with an abnormal chest X-ray, were included in the study. All patients were submitted to clinical, radiological and laboratory evaluation, after which induced sputum and bronchoscopy with bronchoalveolar lavage were carried out. The samples were subjected to the following techniques: Gram and Ziehl-Neelsen staining, quantitative culture growth for pyogenic bacteria, direct staining for fungi, culture growth for mycobacteria and fungi, and Grocott-Gomori staining for Pneumocystis jiroveci, as well as total and differential cell counts. The samples with P. jiroveci were selected, as well as the samples for which no etiologic agents were observed. Forty-five patients with a mean age of 34.6, 38 male and 40 Caucasian, comprised the subjects. Interstitial infiltrate was the most frequent radiological pattern (53.3%). The induced sputum sensitivity was 58.8%, specificity 81.8%, predictive positive value 90.9%, predictive negative value 39.1% and accuracy 64.4%, for the diagnosis of pneumocystosis, compared with BAL. Based on these data, induced sputum is a useful technique for the diagnosis of pneumocystosis in HIV patients.
Journal of Clinical Microbiology
We report on the development of a rapid nested PCR protocol for 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 tested with a P. carinii cyst suspension and was found to be 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 prospectiv...
The Southeast Asian journal of tropical medicine and public health, 2000
Respiratory specimens were prospectively examined for Pneumocystis carinii from 53 patients. The majority of specimens were comprised of expectorated sputum, induced sputum, broncho-alveolar lavage (BAL), and tracheal aspirates. In only four patients Pneumocystis carinii (P. carinii) was detected. All the samples were produced by broncho-alveolar lavage. Candida spp and Aspergillus spp were also identified in a small number of patients. Acid-fast-bacilli were not detected in any of the cases under study. There were no sex-related differences in distribution. The present prospective study was undertaken in order to determine P. carinii infections in human immunodeficiency virus (HIV) seropositive and seronegative individuals. Expectorated sputum samples were probably the major limiting factor in low positivity for detection of P. carinii and study of BAL specimens would be more useful for better results.
Journal of Clinical Pathology, 1994
Aim-To prepare a monoclonal antibody (EB9) against the trophozoite form of Pneumocystis carinii and to test its efficacy for detecting infection with this organism. Method-The sensitivity and specificity of the EB9 antibody were assessed by comparing it with other conventional stains (Papanicolaou, Giemsa and Grocott) and 3F6 antibody in 33 bronchoalveolar lavage specimens from HIV positive patients suspected of having P carinii pneumoma. Results-P carinni infection was detected in 15 of 33 patients. In 14 cases the organism was detected by two or more of the staining methods used; however, EB9 failed to detect infection in two cases which were positive by other staining techniques. In one case P carinii infection was detected by EB9 only. Conclusion-The results of this study suggest that P carinii infection in the lung may occur in two forms: the cyst form and the trophozoite form, which may explain the observed variation in response to treatment.
Clinical infectious …, 2000
(PCR) analysis of respiratory specimens has been investigated as a rapid diagnostic method. We have previously reported on the utility of this technique for diagnosing PCP in HIVinfected patients. In this report we evaluate PCR used in a blinded study design to avoid biases inherent to retrospective and nonblinded studies. The diagnosis of PCP was established on the basis of clinical findings and morphological studies of bronchoalveolar lavage (BAL) and/or lung biopsy specimens before PCR testing. PCR was performed without knowledge of the diagnosis. PCR results were graded from "negative" to 3+ on the basis of intensity of the banding pattern. Forty-seven patients were enrolled in the study, including 18 with proven PCP and 29 with other conditions. PCR was positive at grade 1 or higher for all 18 patients with PCP (100% sensitivity), at grade 2 or higher for 13 patients (72.2% sensitivity), and at grade 1 or higher for 4 of the 29 control patients (specificity of 86.2%). If a grade 2 or higher was required for diagnosis, the specificity improved to 100%. Results were reproducible with testing of a second aliquot for 46 of 47 patients (97.8%). Our findings confirm that PCR is a sensitive and reproducible test for detection of P. carinii in BAL specimens. Problems with false-positive results for control patients, however, limit the applicability of this method.
Clinical Infectious Diseases, 2000
(PCR) analysis of respiratory specimens has been investigated as a rapid diagnostic method. We have previously reported on the utility of this technique for diagnosing PCP in HIVinfected patients. In this report we evaluate PCR used in a blinded study design to avoid biases inherent to retrospective and nonblinded studies. The diagnosis of PCP was established on the basis of clinical findings and morphological studies of bronchoalveolar lavage (BAL) and/or lung biopsy specimens before PCR testing. PCR was performed without knowledge of the diagnosis. PCR results were graded from "negative" to 3+ on the basis of intensity of the banding pattern. Forty-seven patients were enrolled in the study, including 18 with proven PCP and 29 with other conditions. PCR was positive at grade 1 or higher for all 18 patients with PCP (100% sensitivity), at grade 2 or higher for 13 patients (72.2% sensitivity), and at grade 1 or higher for 4 of the 29 control patients (specificity of 86.2%). If a grade 2 or higher was required for diagnosis, the specificity improved to 100%. Results were reproducible with testing of a second aliquot for 46 of 47 patients (97.8%). Our findings confirm that PCR is a sensitive and reproducible test for detection of P. carinii in BAL specimens. Problems with false-positive results for control patients, however, limit the applicability of this method.