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PubMed2005

[The expression of survivin messenger RNA in sputum and cancerous tissue in human lung cancer].

OBJECTIVE: To study the diagnostic significance of the expression of survivin mRNA in transbronchial biopy samples and sputum samples in lung cancer. METHODS: The resected lung cancer tissues and their para-carcinomatous normal tissue specimens of 41 patients with lung cancer and tissue specimens of 9 patients with benign pulmonary diseases were studied. 110 bronchial biopsy specimens from 80 patients with lung cancer and 30 patients with benign pulmonary diseases and their 160 sputum samples were also evaluated. RT-PCR was performed for the detection of survivin mRNA expression in specimens. The results were compared with their histological or cytological examinations. RESULTS: (1) The positive rate of survivin mRNA in resected lung cancer tissues (29/41; 70.7%) was higher than that in the para-carcinomatous normal tissues (7/41; 17.1%) and the benign pulmonary tissues (1/9; 11.1%, P < 0.05). There was no statistical difference (P > 0.05) in the positive rate of survivin mRNA between para-carcinomatous normal tissues and the benign pulmonary disease tissues. In the bronchial biopsy samples, the positive rate of survivin mRNA in 80 lung cancer tissues (51/80; 63.8%) was also higher than that in the benign pulmonary tissues (4/30; 13.3%, P < 0.05). There were no relationships between survivin mRNA expression and age, sex, histopathological subtype, differentiation, or lymph node metastases of the patients. The sensitivity and specificity of diagnosis for lung cancer by detecting survivin mRNA in resected cancer tissue and transbronchial biopsy tissue were 63.8% - 70.7% and 86.7% - 88.9%, respectively. (2) The sensitivity of cytological examinations combined with detecting survivin mRNA in sputum samples was higher than that of either cytological examination or survivin mRNA detection of sputum samples alone (P < 0.05). The sensitivity of the diagnosis for lung cancer increased from 47.1% (sputum cytology alone) to 80.2% (sputum survivin mRNA detection combined with sputum cytology, P < 0.05) and the negative predictive value increased from 37.9% for sputum cytology alone to 57.9% (P < 0.01) for sputum survivin mRNA detection combined with sputum cytology. The specificity did not change significantly. CONCLUSIONS: Detecting survivin mRNA expression from bronchial biopsy specimens might be used as one of the specific molecular markers for diagnosis of lung cancer, while the detection of survivin mRNA from sputum samples as a new ancillary diagnostic method for lung cancer.

Adult↗
PubMed2006

[Number of sputum cultures by MGIT system needed to diagnose pulmonary tuberculosis].

PURPOSE: To study the number of sputum cultures by MGIT system (Becton-Dickinson) needed to diagnose pulmonary tuberculosis. OBJECT AND METHOD: Prospective study of all patients who visited our hospital, and were strongly suspected of pulmonary tuberculosis during the period from Jan. 2002 to Sept. 2003. In these patients, 3 consecutive sputum cultures were done by both MGIT-system and egg-based Ogawa medium (1 slant). RESULTS: Altogether 290 cases of sputum-culture positive pulmonary tuberculosis were available for analysis. In 210 first-sputum-smear positive cases, incremental yield of 3rd sputum culture in 3 consecutive MGIT cultures was equal to or less than 1.0% and 98.1% of culture positive cases were detected by 2 consecutive MGIT cultures. In 80 first-sputum-smear negative cases, incremental yield of 3rd sputum culture in 3 consecutive MGIT cultures was equal to or more than 5.0%, and 90.0% of culture positive cases were detected by 2 consecutive MGIT cultures. This detection rate was almost the same as the calculated detection rate (91.4%) by 3 consecutive Ogawa (2 slant) cultures (previous standard method). CONCLUSION: It was suggested that 2 consecutive sputum cultures by MGIT were sufficient to detect M. tuberculosis in first-sputum-smear positive cases, but 3 consecutive sputum cultures by MGIT were relatively useful in first-sputum-smear negative cases.

Culture Media↗
PubMed2006

[Expression of survivin mRNA of sputum and pleural effusions in human lung cancer].

OBJECTIVE: To determine the diagnostic value of the expression of survivin mRNA in sputum samples and pleural effusions in lung cancer. METHODS: The sputum samples of 104 patients with lung cancer and 30 patients with chronic obstructive pulmonary disease (COPD), and the pleural effusion of 56 patients with lung cancer and 30 patients with tuberculosis pleural effusions were detected.Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the survivin mRNA expression in the specimens. The results were compared with their cytological examinations. RESULTS: The sensitivity of the cytological examinations combined with the detection of survivin mRNA in sputum samples was higher than that of either cytological examination or survivin mRNA detection of sputum samples alone (P<0.01). The sensitivity of the diagnosis for lung cancer increased from 37.5% (sputum cytology alone) to 78.8% (sputum survivin mRNA detection combined with sputum cytology) (P<0.01), and the negative predictive value increased from 31.6% (sputum cytology alone) to 43.5% (sputum survivin mRNA detection combined with sputum cytology) (P<0.01). The sensitivity of the cytological examinations combined with the detection of survivin mRNA in pleural effusion samples was higher than that of cytological examination of pleural effusion samples alone (P<0.01). The sensitivity of the diagnosis for lung cancer increased from 42.9% (pleural effusion cytology alone) to 80.4% (pleural effusion survivin mRNA detection combined with cytology) (P<0.01), and the negative predictive value increased from 48.4% (pleural effusion cytology alone) to 77.8% (pleural effusion survivin mRNA detection combined with cytology) (P<0.01). CONCLUSION: The detection of survivin mRNA from sputum samples and pleural effusions samples is a new diagnostic method for lung cancer.

Adolescent↗
PubMed2004

Assessment of the usefulness of sputum culture for diagnosis of community-acquired pneumonia using the PORT predictive scoring system.

BACKGROUND: The usefulness of sputum culture in guiding microbiological diagnosis of community-acquired pneumonia is controversial. We evaluate and assess it using the Patients Outcome Research Team (PORT) predictive scoring system. METHODS: A cohort of 1669 patients with community-acquired pneumonia was studied. Before administering antibiotic therapy, sputum was collected and its quality evaluated. Samples were gram stained and those of good quality were assessed for a predominant morphotype (PM). Sputum cultures were processed according to standard protocols. RESULTS: A sputum sample was obtained from 983 (59%) of the 1669 patients and 532 (54%) of the samples were of good quality. There was a PM in 240 (45%) of the latter samples (ie, for 14.4% of the 1669 patients) and there was no PM in 292 (55%). Culture yielded a microorganism in 207 (86%) of the 240 samples with PM and 57 (19.5%) of the 292 samples without PM (P<.05). Rates of sputum obtained, good-quality sputum specimens, PM identification, and positive culture were not significantly different among the PORT-score groups of patients (P>.05). The sensitivity and specificity of the gram-positive diplococci identification in the sputum culture of Streptococcus pneumoniae were 60% and 97.6%, and the positive and negative predictive values were 91% and 85.3%, respectively. CONCLUSIONS: Good-quality sputum with PM could be obtained in only 14.4% of all patients. A PORT-score group in which sputum could be of greater usefulness in identifying the causative organism could not be identified. The presence of gram-positive diplococci in gram-stained sputum culture was highly specific for S pneumoniae.

Adolescent↗
PubMed2004

Safety and application of induced sputum analysis in childhood asthma.

BACKGROUND: The value of sputum induction in pediatric asthma lies in its potential to directly and noninvasively assess airway inflammation in children, because bronchoscopy and biopsy carry some risk. The Childhood Asthma Management Program (CAMP) study was designed to evaluate the long-term effects of budesonide and nedocromil compared with placebo in children with mild to moderate asthma across 8 centers. OBJECTIVE: At the Denver CAMP site, we sought to evaluate the safety of sputum induction, to determine differences in airway inflammation between treatment groups by using induced sputum analysis, and to examine correlations between other biomarkers and sputum eosinophils. METHODS: Sputum induction was performed, and exhaled nitric oxide, circulating eosinophil counts, and serum eosinophil cationic protein were obtained at treatment discontinuation and after washout. Spirometry and a methacholine challenge were also performed according to the CAMP protocol. RESULTS: Ninety of 117 children provided an adequate sputum sample for analysis. In 9 subjects (3 nedocromil and 6 placebo), sputum induction resulted in bronchospasm. These subjects had greater disease severity, as measured by a lower median prebronchodilator FEV 1 percentage predicted (85.0% vs 96.0%; P =.024) and FEV 1 /FVC ratio (70.0% vs 79.0%; P =.0008); greater bronchodilator reversibility (16.5% vs 6.8%; P =.004); higher serum IgE (1390.0 vs 495.0 ng/mL; P =.017) and circulating eosinophil count (757.0 vs 282.0/mm 3; P =.04); greater use of prednisone (1.9 vs 0.9 courses per 100 person-years; P =.05); and greater supplemental inhaled steroid doses (85.3 vs 0 mg; P =.016). At treatment discontinuation, budesonide-treated patients had a lower median (1st, 3rd quartile) sputum percentage eosinophil (SPEos) (0.2% [0%, 1.2%] vs 0.8% [0.2%, 4.6%]; P =.03) compared with those treated with placebo; no significant difference was noted between nedocromil- and placebo-treated patients. Higher SPEos at the time of treatment discontinuation was associated with asthma worsening that required rescue prednisone (n = 23) during the washout period compared with patients who remained stable (3.6% [0.4%, 6.4%] vs 0.6% [0.2%, 3.2%] SPEos; P =.023). Finally, greater SPEos was associated with atopy, higher bronchodilator reversibility, lower FEV 1 /FVC ratio, higher exhaled nitric oxide levels, circulating eosinophils, sputum and serum eosinophil cationic protein, more prednisone courses during the treatment period, and greater asthma severity. CONCLUSIONS: Sputum induction is a relatively noninvasive and safe procedure that can provide information on eosinophilic inflammation and treatment response and is also associated with several measures of asthma control. However, this procedure still remains a research tool in asthma because of its requirements for technical expertise.

Adolescent↗
PubMed1994

Analysis of cellular and biochemical constituents of induced sputum after allergen challenge: a method for studying allergic airway inflammation.

To determine whether analysis of the constituents of induced sputum permits detection of changes provoked by aerosolized antigen challenge, we performed sputum induction (20-minute inhalation of aerosolized 3% saline solution) before and after aerosolized allergen challenge in eight subjects with asthma. Total cell counts and cell differentials of nonsquamous cells in induced sputum samples were determined after the samples were homogenized in dithiothreitol. Centrifugation of the entire homogenized sputum sample yielded supernatant that could be analyzed for biochemical constituents. We found that the median percentage of eosinophils and neutrophils in induced sputum samples was significantly higher 4 hours after allergen challenge neutrophils in induced sputum samples was significantly higher 4 hours after allergen challenge than at baseline (12% vs 0.5%, p < 0.05; 30.5% vs 7.5%, p < 0.05) and remained high 24 hours after challenge. Median levels of eosinophil cationic protein and histamine in induced sputum supernatants were significantly higher 4 hours after challenge than at baseline (151.3 vs 39.8 ng/ml, p < 0.05; 19.4 vs 8.8 micrograms, p < 0.05) and remained significantly higher 24 hours after challenge. Tryptase was detectable in sputum from seven of the subjects, and in these subjects, we found a trend toward an increase in median tryptase levels 4 hours after allergen challenge (4.4 vs 2.2 U/L, p = 0.09). We conclude that analysis of induced sputum after aerosolized allergen challenge reveals changes in inflammatory cells and markers similar to those reported in bronchoalveolar lavage fluid and that sputum induction is a useful noninvasive method for studying allergic airway inflammation in asthma.

Administration, Inhalation↗
PubMed1999

Nitric oxide metabolites in patients with asthma: induced sputum versus blood.

Nitric oxide (NO) plays an important role in physiological regulation of the airways. The monitoring of airway inflammation has being observed in bronchial asthma directly, by sputum examination, and indirectly, by measurements in peripheral blood. To investigate the diagnostic value of these two methods, we compared NO metabolites in induced sputum and serum obtained in patients with asthma and control subjects. Hypertonic saline induced sputum and serum were obtained in 13 patients with asthma and 10 control subjects. NO metabolite level was assayed by using modified Griess reaction. Eosinophil cationic protein (ECP) was measured by fluoroimmunoassay, and detected interleukin (IL)-5 by a sandwich ELISA. The accuracy of the tests was measured by plotting the data in receiver operating characteristic (ROC) curves and comparing the area under the curve for NO metabolites. Asthmatic patients, compared with control subjects, had significantly higher NO metabolites in induced sputum (1252.5+/-203.3 mol l(-1) vs. 557.2+/-101.5 mol l(-1), P<0.01) but not in serum. IL-5 in induced sputum was detected more frequently in patients with asthma than in control subjects [11/13 (84.6%) vs. 1/10 (10%), P<0.01]. Asthmatic patients, compared with control subjects, had significantly higher ECP concentration in induced sputum (1270.0+/-197.9 g l vs. 154.6+/-47.4 g l(-1), P<0.01). There were significant positive correlations between NO metabolites in induced sputum and eosinophils, ECP in induced sputum (r=0.58 P<0.05; r=0.64, P<0.01) in patients with asthma but not in serum. The area under the ROC curve showed that NO metabolites in induced sputum (0.78) are more accurate marker than NO metabolites in serum (0.53) (P<0.05). These findings suggest that NO metabolites in induced sputum is a more valuable indicator to monitor asthmatic airway inflammation than those in serum.

Adult↗
PubMed2002

Eosinophils and eosinophilic cationic protein in induced sputum and blood: effects of budesonide and terbutaline treatment.

BACKGROUND: There is a need for easily measurable markers of airway inflammation to guide the use of anti-inflammatory treatment in asthma. Eosinophilic cationic protein (ECP) levels in sputum and blood correlate with clinical severity, and serial measurements of ECP have been proposed as a suitable candidate. AIMS AND METHODS: Our aim was to confirm that sputum and serum ECP measurements would provide a more sensitive indicator of responses to asthma treatment than eosinophil counts per se, in a randomized, placebo-controlled, crossover study of terbutaline, budesonide, and their combination in patients with chronic persistent asthma. We compared the changes in eosinophil counts and ECP in induced sputum and blood during each treatment period. RESULTS: Budesonide and combined treatment caused a significant reduction in sputum eosinophils (-2.7% and -2.3%, respectively, P < 0.05). Sputum eosinophils increased with terbutaline (+3.9%, P = 0.049). In contrast, the changes for sputum ECP were not significant. There was a similar treatment effect on blood eosinophils, but not for serum ECP. Correlations between sputum and blood eosinophils were significant with and without budesonide, but were nonsignificant between sputum and blood ECP during the active treatments. Correlations between sputum eosinophils and ECP, and between blood eosinophils and serum ECP were greatest during treatment with placebo or terbutaline alone: budesonide weakened or abolished these relationships. CONCLUSIONS: Compared with eosinophil counts, ECP measurements in either induced sputum or serum failed to reflect treatment-related changes in chronic asthma. We conclude that ECP is not a sensitive or reliable means of evaluating airway inflammation, and can not be recommended for assessing responses to anti-inflammatory therapy.

Administration, Topical↗
PubMed1999

Relationship between induced sputum cell counts and fluid-phase eosinophil cationic protein and clinical or physiologic profiles in mild asthma.

BACKGROUND: Sputum analysis is the only non-invasive method to examine airway inflammatory processes in subjects with asthma. The aim of this study was to investigate the relationship between cell counts and fluid phase levels in induced sputum in subjects with mild asthma, and the severity of asthma as assessed by clinical, physiologic and blood measurements. METHODS: Forty patients with mild asthma, aged 17 to 49 years were studied (good sputum sample only from 31). On the first day, spirometry and methacholine challenges were performed. After 2 to 4 days, venous blood for absolute eosinophil count and eosinophil cationic protein (ECP) measurement was obtained and sputum was induced by inhalation of hypertonic saline. For the next 15 days subjects recorded their peak expiratory flow (PEF), symptom scores, and beta2-agonist requirements twice daily. Differential counts of leukocytes were done on cytospin preparations of homogenized sputum and the supernatant was examined for eosinophil cationic protein (ECP). RESULTS: Sputum eosinophil counts and not neutrophil, epithelial cells, macrophages, or lymphocytes, were inversely correlated to FEV1/FVC % (r = -.57, P = .0008) and to PC20-methacholine (r = -.40, P = .024). No statistical relationship was obtained between eosinophil counts and either symptom scores, bronchodilator requirements, or daily PEF variability. Sputum ECP values were correlated to FEV1/FVC% (r = -.41, P = .026) but not to PC20 (r = -.32, P = .08) or clinical scores or PEF variation. A trend to significance was appreciated between peripheral blood and sputum eosinophil counts (r = .34, P = .067) and no relationship was found between sputum and serum ECP values (r = .10, P = .38). CONCLUSIONS: Although sputum markers give some information about disordered lung function and physiologic changes in the airways, they are not the only factors concerned in the clinical expression of mild asthma.

Adolescent↗
PubMed2000

Diagnostic accuracy of sputum outcomes in chronic stable asthma.

BACKGROUND: Asthma with non-remitting airflow obstruction may not always be differentiated from COPD with airway hyperreactivity. Many attempts have been made to find useful markers for the distinction between these two disorders. OBJECTIVE AND METHODS: In order to help the finding of a useful marker for the diagnosis of asthma in the population of patients with airway obstruction we analysed the diagnostic accuracy of sputum eosinophils and sputum ECP in 91 patients with asthma, 15 patients with chronic bronchitis, 32 patients with chronic obstructive pulmonary disease (COPD) and 20 controls subjects, by performing ROC analysis. RESULTS: Sputum eosinophils were above the normal range of our laboratory (0-3.7%) in 48 asthma patients and in six COPD patients, while sputum ECP (normal range < 85 microg/L) was high in 65 asthma patients, in 24 COPD patients and in nine chronic bronchitis patients. The ROC analysis revealed that sputum eosinophils count (AUC = 0.82) was more accurate than both sputum ECP levels (AUC = 0.56) (P < 0.0001) and beta2-reversibility (AUC = 0.53) (P = 0.0001) in differentiating asthmatic from non-asthmatic subjects (COPD, chronic bronchitis patients and normal subjects). The diagnostic accuracy of ECP was similar to that of bronchial reversibility (P = 0.76). When ROC analysis was performed by including only patients with airway obstruction (36 asthmatics with airway obstruction and COPD patients), both eosinophil count (AUC = 0.77) and beta2-reversibility (AUC = 0.66) were more accurate than ECP measurement (AUC = 0.39) in discriminating asthmatics from COPD patients (P < 0.00001 and P = 0.04, respectively). CONCLUSION: Sputum eosinophils seem to be valid markers for detecting asthma in a population of patients with airway obstruction. Moreover, the higher diagnostic accuracy of eosinophils in the sputum compared to sputum ECP and bronchial reversibility reinforces the role of cytological analysis of sputum in the diagnosis of chronic stable bronchial asthma.

Adolescent↗
PubMed2002

Sputum processing: a new method to improve cytospin quality.

BACKGROUND: Sputum examination is being increasingly used as a non-invasive method for studying airway inflammation. However, the application of sputum still presents some methodological problems and the results of sputum analysis may be substantially flawed by salivary contamination, cell and mucus debris. In addition, much work is needed to deepen the possibility of extensive application of cell and molecular biology techniques to sputum analysis. OBJECTIVE: In an attempt to improve the technique of sputum processing, we investigated the effect of: (i) 20 and 11 microm filtration in addition to 40 microm on salivary contamination; (ii) Percoll density gradient centrifugation on sputum slides quality; (iii) a culture medium (Minimum Essential Medium containing HEPES 22 mm, pH 7.4: MEM) as washing and suspension solution compared to PBS on cell viability. METHODS: Induced sputum samples were obtained in 37 asthmatics. 21 samples were processed as selected sputum and 16 samples as entire expectorates. After dithiotreitol (DTT) homogenization, each specimen was aliquoted in two parts of equal volume. One portion was processed with the usual method, the other using a modified method: cell pellet was suspended in sterile MEM, filtered through 40, 20 and 11 microm net filters and separated from the residual debris by Percoll gradient centrifugation. RESULTS: As compared to the current sputum processing this method resulted in: (i) no selective bronchial cellular loss; (ii) a significant decrease of salivary contamination, particularly in entire expectorates in which squamous cells were reduced from 47 (36) to 15.5% (20) as median values and interquartile range; (iii) a higher proportion of good quality cytospins; (iv) maintenance of cell viability over the time (88% vs. 81% in MEM and PBS, respectively) 1 h after sample collection. CONCLUSION: In the present study we demonstrated that the proposed method is feasible and makes it possible to overcome most of the technical limits met with the commonly used method, pointing to a potential extension of induced sputum application for more sophisticated techniques.

Adolescent↗
PubMed1999

Characterization of immunoglobulin binding factor in sputum from patients with chronic airway diseases.

OBJECTIVE: Immunoglobulin binding factor (IgBF) is known to bind immunoglobulin, to interact with anti-Fc gamma-III antibodies and to be present in the lower respiratory tract of normal healthy subjects. In this study, in order to clarify the role of IgBF in respiratory diseases, we investigated whether IgBF exists in the airway of patients with chronic airway diseases. METHODOLOGY: IgBF was measured in the sputum of 28 normal subjects and 59 patients with chronic airway diseases including 37 cases of chronic bronchitis, 18 bronchiectasis, and four diffuse panbronchiolitis by enzyme-linked immunosorbent assay. RESULTS: Immunoglobulin binding factor concentration in the sputum of patients with chronic airway diseases (purulent sputum, 50.2 +/- 8.2 microg/mL; mucoid sputum, 88.6 +/- 12.8 microg/mL) was higher than that in induced sputum of normal subjects (6.3 +/- 5.5 microg/mL; P < 0.001). Immunoglobulin binding factor level in mucoid sputum was significantly higher than that in purulent sputum (P < 0.05). A significant inverse correlation was shown between the IgBF level and the elastase activity in sputum, and the concentration of IgBF purified from seminal plasma was decreased by treatment with neutrophil elastase, indicating that IgBF might be degraded by elastase. In the gel filtration chromatography of both types of sputum, IgBF was eluted in a region corresponding to a molecular weight of 27 kDa as a single peak. Western blot analysis with a monoclonal antibody to IgBF indicated that IgBF in both types of sputum had a molecular weight of 27 kDa under non-reducing conditions and of 16 kDa under reducing conditions. CONCLUSION: These results demonstrate that a high level of IgBF is present in the respiratory tract of patients with chronic airway diseases and may be related to the pathogenesis of these diseases.

Adult↗
PubMed2002

Sputum induction problems identified through genetic fingerprinting.

OBJECTIVE: To identify the contamination source of a cluster of eight positive Mycobacterium tuberculosis isolates from one laboratory session. METHODS: Spoligotyping was performed on M. tuberculosis isolates processed during one laboratory session. Laboratory and sputum induction protocols and records were reviewed. Sputum induction staff were interviewed. An environmental assessment of the sputum induction booth was performed. RESULTS: Spoligotyping identified a unique strain of susceptible M. tuberculosis from five induced sputa collected at Clinic A on the same day. Three specimens processed concurrently from other clinics had spoligotypes different from each other and from the cluster strain. A laboratory investigation revealed no procedural lapses. Sputum induction records from Clinic A indicated that patient 1 in the sputum induction booth had prior culture-confirmed tuberculosis. Patient 2 had a history of a drug-resistant strain. Patient 3 had completed tuberculosis treatment, with positive cultures 7 months earlier. Patients 4 and 5 were new to the clinic and had no subsequent positive M. tuberculosis specimens. The sputum induction booth was working within normal parameters. Sputum induction that day was overseen by a new employee with limited training and no supervision. A review of the sputum induction protocol identified ambiguity regarding care of the ultrasonic nebulizer between patients, which may have led to reuse of the discarded nebulizer solution from patient 1. CONCLUSIONS: A break in the sputum induction protocol may have contributed to contamination of patient specimens. Sputum induction is complicated, mandating adequate staff training and supervision and patient preparation. Spoligotyping identified a potential source of M. tuberculosis contamination.

Clinical Laboratory Techniques↗
PubMed1994

Use of induced sputum specimens for microbiologic diagnosis of infections due to organisms other than Pneumocystis carinii.

The optimal diagnostic approach to pneumonia provides a rapid microbiologic identification of pulmonary pathogens by the least invasive means. The technique of sputum induction has been useful in the evaluation of patients with Pneumocystis carinii or mycobacterial pneumonia. It is not known whether induced sputum samples are preferable for the detection of pathogens other than P. carinii or mycobacteria. Microbiologic yields were evaluated from identically processed induced and conventional sputum samples collected from 509 consecutive patients. No statistically significant differences were found between the microbiologic yields of induced and spontaneous sputum samples. Bacterial pathogens were isolated in 19.6% of induced and 23.5% of routine specimens. Mycobacteria were cultured from 13.1% of routine and 9.4% of induced specimens. Non-Candida albicans fungi grew from 24% of routine and 20% of induced specimens. The process of sputum induction with aerosolized hypertonic saline did not alter either the purulence or the bacterial quantitation of Gram-stained sputum specimens. Sputum induction has been useful for the cytologic diagnosis of malignancy, for the diagnosis of pneumocystosis and tuberculosis, and in patients unable to spontaneously produce sputum samples. The use of induced sputum samples for the diagnosis of other infections may not be necessary when routine sputum specimens are available.

Bacteria↗
PubMed1994

Detection of beta lactamase in sputum.

AIMS: To develop a rapid, sensitive, and reproducible test for the detection of beta lactamase in sputum, and to relate these findings to bacteriological culture results. METHODS: One hundred and twenty sputum samples from inpatients were investigated for beta lactamase activity using the chromogenic cephalosporin nitrocefin. Sputum samples were sonicated and incubated aerobically at 37 degrees C with nitrocefin for up to two hours. Positive results (production of a red colour) were treated with blue sepharose beads to remove albumin (which can cause non-specific hydrolysis of nitrocefin) and retested. Samples were also cultured for both aerobic and anaerobic bacteria, with all isolates being tested for beta lactamase production using nitrocefin. All positive sputum samples and beta lactamase producing isolates were further examined by isoelectric focusing (IEF) to determine isoelectric point(s) (pIs). RESULTS: The process of sonication and albumin removal had no demonstrable effect on beta lactamase activity. Forty seven of the 120 sputum samples were positive on initial testing, and of these, 16 remained positive following removal of albumin. These 16 subsequently yielded 19 beta lactamase producing bacteria. All sputum samples yielding beta lactamase producing bacteria were also positive on direct nitrocefin testing. On no occasion were sputum samples positive in the absence of enzyme producing bacteria--that is, the test was both 100% specific and 100% sensitive. The presence of beta lactamase activity in the sputum samples was also confirmed using a microbiological method. In 11 sputum samples the beta lactamases detected had similar pI values to the beta lactamases obtained from their bacterial isolates. CONCLUSION: Detection of beta lactamase activity in sputum using nitrocefin, after treatment with blue sepharose beads, is a rapid, reproducible test with high specificity and sensitivity. Analytical isoelectric focusing showed that for 11 of the 16 positive sputa, the source of the beta lactamases could be traced to their concurrent bacterial isolates.

Bacteria↗
PubMed2000

Correlation of CD4:CD8 ratio and tumour necrosis factor (TNF)alpha levels in induced sputum with bronchoalveolar lavage fluid in pulmonary sarcoidosis.

BACKGROUND: An increased CD4:CD8 lymphocyte ratio and raised cytokine levels in bronchoalveolar lavage (BAL) fluid are characteristic of pulmonary sarcoidosis. Sputum induction has been used as a non-invasive tool for investigating the airways and may be useful in investigating inflammation in patients with sarcoidosis in whom endobronchial, peribronchial, and parenchymal inflammation is present. This study aimed to correlate the total and differential cell counts, CD4:CD8 ratio, and tumour necrosis factor (TNF)alpha levels between induced sputum and BAL fluid in patients with pulmonary sarcoidosis. METHODS: Fourteen patients with newly diagnosed biopsy proven sarcoidosis and six healthy controls were investigated. Sputum induction and BAL was carried out at the initial visit and repeated following six months of treatment with oral prednisone. RESULTS: There was no correlation of differential cell counts between induced sputum and BAL fluid. The CD4:CD8 ratio in induced sputum correlated strongly with that in BAL fluid (5.5 (0. 4:1) versus 4.4 (0.2:1); r = 0.8, p<0.001) and the fall in the ratio following six months of treatment in sputum paralleled that in BAL fluid (3.4 (0.2:1) versus 2.4 (0.2:1)). The TNF alpha levels in sputum also correlated with levels in the BAL fluid (11.9 (1.5) pg/ml versus 17.6 (2.7) pg/ml; r = 0.8, p<0.001). The fall in sputum TNF alpha levels following six months of treatment paralleled the fall in BAL fluid levels (6.7 (0.9) pg/ml versus 11.6 (1.3) pg/ml). CONCLUSIONS: The CD4:CD8 ratio and TNF alpha levels in induced sputum correlated with those in BAL fluid and paralleled changes with treatment. Induced sputum may therefore be a non-invasive surrogate for certain parameters in BAL fluid in patients with sarcoidosis.

Adult↗
PubMed2000

Induced sputum in children: feasibility, repeatability, and relation of findings to asthma severity.

BACKGROUND: The collection of induced sputum provides a non-invasive method of investigating airway inflammation. Few studies have been performed in children, so a study was undertaken to determine the feasibility of sputum induction, the repeatability of eosinophil counts and sputum eosinophil cationic protein (ECP) levels, and the relation of these to current asthma severity. For comparison, serum ECP levels were also measured. METHODS: In a cross sectional study of children aged 5-15 years, 27 healthy children and 60 with asthma underwent sputum induction using inhaled nebulised hypertonic saline. The whole sputum sample was used for analysis. Ten children with stable asthma repeated the procedure within 10 days. RESULTS: A satisfactory sample (>500 non-squamous cells) was obtained in 61% of children with asthma and in 60% of healthy controls. The limits of agreement within subjects ranged from a 0.68 to 2.8 fold difference for eosinophil differential counts and from 0.38 to 4.4 fold for sputum ECP. Despite a median of 42% squamous cells, significant differences were found between asthma and healthy controls for the eosinophil differential count (p = 0.0004), total eosinophil counts (p = 0.03), and sputum ECP level (p = 0.0001). Overall, there was no correlation between any marker of airway inflammation and asthma severity, however expressed, including lung function. CONCLUSIONS: Sputum induction is only possible in a proportion of children. The repeatability of sputum cell counts and ECP levels, measured in a small number of children, was similar to that reported in adults. Sputum analysis revealed no evidence of airway inflammation in a number of highly symptomatic children with asthma.

Adolescent↗
PubMed1995

Exacerbations of asthma without sputum eosinophilia.

BACKGROUND: Sputum analysis provides a non-invasive method of examining the airway secretions of subjects with asthma in order to better understand the inflammatory process. Increased proportions of eosinophils are generally seen in the sputum of subjects with asthma, especially when there is an exacerbation. An unexpected observation in the sputum of subjects with mild exacerbations of asthma is reported. METHODS: Thirty four consecutive subjects with symptoms consistent with a mild exacerbation of asthma were recruited for a treatment study. Inclusion criteria required persistent symptoms of chest tightness, dyspnoea, or wheezing for two weeks (without spontaneous improvement or alteration in dose of inhaled corticosteroid) and a forced expiratory volume in one second (FEV1) that was reversible to more than 75% predicted or known best to ensure the exacerbation was mild. Sputum (spontaneous or induced with hypertonic saline) from all subjects was examined for differential cell counts. Eosinophilic sputum was defined as > or = 4% eosinophils on two occasions or > 10% eosinophils once. Clinical characteristics, sputum differential counts, and measurements of airways obstruction were compared between the subjects with and without sputum eosinophilia. RESULTS: Almost half of the subjects (16 of 34) considered to have mildly uncontrolled asthma had no sputum eosinophilia. In comparison with the subjects who had sputum eosinophilia the non-eosinophilic group had less airways obstruction (FEV1% predicted 88% v 70%) and less severe airways hyperresponsiveness (PC20 methacholine 0.45 mg/ml v 0.13 mg/ml). There was no difference between the groups in the type or prevalence of symptoms, history of recent infections, smoking, relevant allergen exposure, or use of inhaled corticosteroid. CONCLUSIONS: Symptoms of mildly uncontrolled asthma are not always associated with eosinophilic airways inflammation as measured by sputum analysis. The causes and treatment of the non-eosinophilic condition require further investigation.

Adolescent↗