The agreement between bronchoalveolar lavage, bronchial wash and sputum culture: a retrospective study.

PURPOSE: Bronchoalveolar lavage is commonly used in clinical practice for unresolved pneumonia. However, bronchoalveolar lavage is not suitable for all patients as it is an invasive procedure and can worsen oxygenation. The diagnostic value of bronchial wash and sputum has been debated extensively over the years. In this study, we aim to compare the diagnostic value in several pathogens of bronchoalveolar lavage and bronchial wash, and secondarily bronchoalveolar lavage and sputum. METHODS: We retrospectively included all adult patients in our hospital who underwent bronchoalveolar lavage, bronchial wash, and where sputum sampling was done between January 1st of 2018 and December 31st of 2021. The intraclass correlation coefficient was computed for the three tests. RESULTS: In total, 308 patients were included. We found a level of correlation of 0.819 and 0.865, respectively, between bronchoalveolar lavage and bronchial wash for two pathogens: Staphylococcus aureus and Pseudomonas aeruginosa. For Stenotrophomonas maltophilia and Aspergillus fumigatus, we found an intraclass correlation coefficient of 0.568 and 0.624, respectively. Between bronchoalveolar lavage and sputum, we found varying levels of agreement. CONCLUSION: Our study shows reasonably well agreement levels between bronchoalveolar lavage and bronchial wash, suggesting that bronchial wash could potentially be an alternative to bronchoalveolar lavage.

Large airway T cells in adults with former bronchopulmonary dysplasia.

BACKGROUND: Bronchopulmonary Dysplasia (BPD) in infants born prematurely is a risk factor for chronic airway obstruction later in life. The distribution of T cell subtypes in the large airways is largely unknown. OBJECTIVE: To characterize cellular and T cell profiles in the large airways of young adults with a history of BPD. METHODS: Forty-three young adults born prematurely (preterm (n = 20), BPD (n = 23)) and 45 full-term-born (asthma (n = 23), healthy (n = 22)) underwent lung function measurements, and bronchoscopy with large airway bronchial wash (BW). T-cells subsets in BW were analyzed by immunocytochemistry. RESULTS: The proportions of both lymphocytes and CD8 + T cells in BW were significantly higher in BPD (median, 6.6%, and 78.0%) when compared with asthma (3.4% and 67.8%, p = 0.002 and p = 0.040) and healthy (3.8% and 40%, p < 0.001 and p < 0.001). In all adults born prematurely (preterm and BPD), lymphocyte proportion correlated negatively with forced vital capacity (r= -0.324, p = 0.036) and CD8 + T cells correlated with forced expiratory volume in one second, FEV1 (r=-0.448, p = 0.048). Correlation-based network analysis revealed that lung function cluster and BPD-birth cluster were associated with lymphocytes and/or CD4 + and CD8 + T cells. Multivariate regression analysis showed that lymphocyte proportions and BPD severity qualified as independent factors associated with FEV1. CONCLUSIONS: The increased cytotoxic T cells in the large airways in young adults with former BPD, suggest a similar T-cell subset pattern as in the small airways, resembling features of COPD. Our findings strengthen the hypothesis that mechanisms involving adaptive and innate immune responses are involved in the development of airway disease due to preterm birth.

The development, validation, and in vivo testing of a high-precision bronchial epithelial lining fluid sampling device.

Introduction: Analysis of respiratory biomarkers or pharmaceutical drug concentrations in bronchial epithelial lining fluid (bELF) using a high-precision sampling method is crucial for effective clinical respiratory diagnostics and research. Here, we utilized a cellulose matrix as an absorptive probe for bELF sampling, subsequently testing the design of a device and sampling technique in vivo. Methods: The absorptive matrix [Whatman® qualitative filter paper (Grade CF-12)] was first tested through tissue-contact experiments on porcine airway tissue. The absorption and elution capacity of the matrix, as well as the laboratory processing and analysis method, was validated with a range of Interleukin-8 (CXCL8) and C-Reactive protein (CRP) stock solutions. Subsequently, the device's design was optimized for universal in-house production and both, safe and efficient sampling. The airway sampling method was then tested in a group of 10 patients with Chronic Obstructive Pulmonary Disease (COPD). For each patient, a bELF sample was obtained using the newly developed bELF probe, as well as a reference 20 mL saline bronchial wash sample. Supernatants were assessed, using an immunoassay, for levels of the pro-inflammatory markers CXCL8, Myeloperoxidase (MPO), and CRP. The bELF samples were compared to bronchial wash. Results: The Whatman® qualitative filter paper (Grade CF-12) bELF probes adhered to porcine airway tissue, softening slightly upon wetting. The material maintained architectural integrity following the removal of the probes, leaving no residual fibers on the porcine airway mucosa. The bELF probe design was optimized for bronchoscopic delivery and in-house production. On average, a fully saturated bELF probe carried 32 µL of protein-rich fluid. The mean return of CXCL8 and CRP from samples collected from a serial dilution series (1, 5, 10, 20 ng/mL) was 69% (range 48%-87%). The bELF probe detected, on average, 7 (MPO), 14 (CRP), and 59 (CXCL8) times higher equivalent inflammatory protein concentrations in the collected bELF probe samples compared to the bronchial wash. Conclusion: The bELF probe is an effective absorptive technology for high-precision bELF sampling without dilution. With a simple in-house production procedure and bronchoscopic sampling technique, this method can be introduced in any bronchoscopic center for a consistent sampling of bELF.

Role of Bronchoscopic Cytology in Diagnosis of Pulmonary Lesions.

Objectives: Respiratory cytology specimens such as bronchoalveolar lavage (BAL) and bronchial wash (BW) obtained using a fiberoptic bronchoscope are very useful in detecting or ruling out various inflammatory conditions, infections, and neoplastic lesions. A study was carried out to determine the usefulness of respiratory cytology in the diagnosis of pulmonary lesions and the limitations of cytology if any, and correlate the results of cytology with biopsies wherever possible. Methods: All bronchoscopic cytology and biopsy specimens received at the pathology laboratory of this tertiary care institute between June 2014 and May 2017 were analyzed. Cytology smears were stained with Leishman's stain, hematoxylin and eosin (H and E), Papanicolaou (PAP), and Ziehl-Neelsen (ZN) stain for all cases and special stains wherever needed. Slides prepared from biopsy specimens were stained with H and E. Immunohistochemistry was used for confirmation and further typing of malignant lesions and diagnosis rendered was compared with the corresponding cytology diagnosis. Results: A total of 120 specimens of BAL or BW cytology with or without biopsy were analyzed. Thirty-three were diagnosed as non-specific inflammatory lesions. The most common malignancy diagnosed by cytology was adenocarcinoma followed by squamous cell carcinoma. Correlating BAL with biopsy specimens, the sensitivity, specificity, and diagnostic accuracy of BAL were 100%, 88.8%, and 91.6%, respectively. Correlating BW with biopsy specimens, the sensitivity, specificity, and diagnostic accuracy of BW were 85.6%, 85.6%, and 85.6%, respectively. Conclusions: Accurate diagnosis can be made from the examination of bronchoscopic cytology specimens in pulmonary inflammation, tuberculosis, fungal infections, and malignancies. Combining respiratory cytology with biopsy and ancillary techniques can aid in better subtyping of neoplastic lesions.

Factors associated with acid-fast bacillus isolation in patients with noncystic fibrosis bronchiectasis: A cross-sectional study.

INTRODUCTION: Acid-fast bacillus (AFB) is a major pathogen that causes noncystic fibrosis bronchiectasis requiring multidrug chemotherapy. Bronchoscopic bronchial wash is performed to determine the causative pathogens of bronchiectasis; but, predictive factors for AFB isolation have not been fully elucidated. This study aimed to determine the factors associated with AFB isolation from bronchial wash samples. METHODS: This was a single-center, cross-sectional study. Patients undergoing bronchoscopic bronchial wash for bronchiectasis were included, whereas those who did not undergo high-resolution computed tomography (HRCT); had acute pneumonia, interstitial lung disease, and a positive polymerase chain reaction result but a negative culture result for AFB; or in whom a guide sheath was used for suspected lung cancer were excluded. Binomial logistic regression was used to analyze the factors associated with a positive culture for AFB. RESULTS: Of the 96 included cases, AFB isolation was observed in the bronchial wash fluid of 26 patients (27%). No smoking history, a positive result for antiglycopeptidolipid (GPL)-core IgA antibody, and the presence of tree-in-bud appearance, multiple granular and nodular images on HRCT were more commonly observed in patients with AFB isolation than in those without. In the multivariate analysis, the tree-in-bud appearance (odds ratio, 4.223; 95% CI, 1.046-17.052) and anti-GPL core IgA antibody (odds ratio, 9.443; 95% CI, 2.206-40.421) were significantly associated with AFB isolation. CONCLUSIONS: The tree-in-bud appearance on HRCT is likely to predict AFB isolation independent of anti-GPL core IgA antibody results. Bronchoscopic bronchial wash should be recommended for bronchiectasis with multiple granulomas on HRCT.

Utility of bile acids in large airway bronchial wash versus bronchoalveolar lavage as biomarkers of microaspiration in lung transplant recipients: a retrospective cohort study.

BACKGROUND: Bronchoalveolar lavage (BAL) is a key tool in respiratory medicine for sampling the distal airways. BAL bile acids are putative biomarkers of pulmonary microaspiration, which is associated with poor outcomes after lung transplantation. Compared to BAL, large airway bronchial wash (LABW) samples the tracheobronchial space where bile acids may be measurable at more clinically relevant levels. We assessed whether LABW bile acids, compared to BAL bile acids, are more strongly associated with poor clinical outcomes in lung transplant recipients. METHODS: Concurrently obtained BAL and LABW at 3 months post-transplant from a retrospective cohort of 61 lung transplant recipients were analyzed for taurocholic acid (TCA), glycocholic acid (GCA), and cholic acid by mass spectrometry and 10 inflammatory proteins by multiplex immunoassay. Associations between bile acids with inflammatory proteins and acute lung allograft dysfunction were assessed using Spearman correlation and logistic regression, respectively. Time to chronic lung allograft dysfunction and death were evaluated using multivariable Cox proportional hazards and Kaplan-Meier methods. RESULTS: Most bile acids and inflammatory proteins were higher in LABW than in BAL. LABW bile acids correlated with inflammatory proteins within and between sample type. LABW TCA and GCA were associated with acute lung allograft dysfunction (OR = 1.368; 95%CI = 1.036-1.806; P = 0.027, OR = 1.064; 95%CI = 1.009-1.122; P = 0.022, respectively). No bile acids were associated with chronic lung allograft dysfunction. Adjusted for risk factors, LABW TCA and GCA predicted death (HR = 1.513; 95%CI = 1.014-2.256; P = 0.042, HR = 1.597; 95%CI = 1.078-2.366; P = 0.020, respectively). Patients with LABW TCA in the highest tertile had worse survival compared to all others. CONCLUSIONS: LABW bile acids are more strongly associated than BAL bile acids with inflammation, acute lung allograft dysfunction, and death in lung transplant recipients. Collection of LABW may be useful in the evaluation of microaspiration in lung transplantation and other respiratory diseases.

Cell Free Methylated Tumor DNA in Bronchial Lavage as an Additional Tool for Diagnosing Lung Cancer-A Systematic Review.

This systematic review investigated circulating methylated tumor DNA in bronchial lavage fluid for diagnosing lung cancer. PROSPERO registration CRD42022309470. PubMed, Embase, Medline, and Web of Science were searched on 9 March 2022. Studies of adults with lung cancer or undergoing diagnostic workup for suspected lung cancer were included if they used bronchial lavage fluid, analyzed methylated circulating tumor DNA, and reported the diagnostic properties. Sensitivity, specificity, and lung cancer prevalence were summarized in forest plots. Risk of bias was assessed using the QUADAS-2 tool. A total of 25 studies were included. All were case-control studies, most studies used cell pellet for analysis by quantitative PCR. Diagnostic sensitivity ranged from 0% for a single gene to 97% for a four-gene panel. Specificity ranged from 8% for a single gene to 100%. The studies employing a gene panel decreased the specificity, and no gene panel had a perfect specificity of 100%. In conclusion, methylated circulating tumor DNA can be detected in bronchial lavage, and by employing a gene panel the sensitivity can be increased to clinically relevant levels. The available evidence regarding applicability in routine clinical practice is limited. Prospective, randomized clinical trials are needed to determine the further usefulness of this biomarker.

International Society for Heart and Lung Transplantation consensus statement for the standardization of bronchoalveolar lavage in lung transplantation.

Bronchoalveolar lavage (BAL) is a key clinical and research tool in lung transplantation (LTx). However, BAL collection and processing are not standardized across LTx centers. This International Society for Heart and Lung Transplantation-supported consensus document on BAL standardization aims to clarify definitions and propose common approaches to improve clinical and research practice standards. The following 9 areas are covered: (1) bronchoscopy procedure and BAL collection, (2) sample handling, (3) sample processing for microbiology, (4) cytology, (5) research, (6) microbiome, (7) sample inventory/tracking, (8) donor bronchoscopy, and (9) pediatric considerations. This consensus document aims to harmonize clinical and research practices for BAL collection and processing in LTx. The overarching goal is to enhance standardization and multicenter collaboration within the international LTx community and enable improvement and development of new BAL-based diagnostics.

Detection of tumor-derived DNA dispersed in the airway improves the diagnostic accuracy of bronchoscopy for lung cancer.

The diagnostic accuracy of bronchoscopy for detecting lung cancer, especially peripheral lung cancer with lesions outside the endoscopically visible range, remains unsatisfactory. The aim of this study was to perform next-generation sequencing on bronchoscopic specimens to determine whether this improves the accuracy of bronchoscopy for diagnosing lung cancer and to identify factors influencing sensitivity. The bronchoscopic sensitivity for diagnosing lung cancer was initially evaluated in 191 patients who underwent lobectomy after bronchoscopy at our hospital. Sputum, bronchial wash fluid, and resected lung cancer specimens were subsequently collected from 18 patients with peripheral small cell lung cancer for genomic analysis. DNA was extracted from formalin-fixed, paraffin-embedded surgical tissue specimens and the supernatant and cell fractions of sputum and bronchial wash fluid. Deep sequencing was performed using a lung cancer panel covering all exons of 53 lung cancer-related genes. The bronchoscopic sensitivity for diagnosing lung cancer at our hospital was 60.7%. Multivariate analysis revealed that this was influenced by tumor size and location, but not histological type or lymph node metastasis. The sensitivity was the highest for biopsy followed by curettage and bronchial wash specimens. DNA mutations homologous to those identified in the primary lesions were detected in the bronchial wash fluid of 10 patients (55.6%), while only 2 patients (11.1%) were diagnosed with lung cancer based on conventional cytological examinations. In conclusion, the addition of genomic analysis to routine pathological examinations improves the diagnostic accuracy of bronchoscopy.

Liquid-based and conventional cytology for bronchial washings/bronchoalveolar lavages in the diagnosis of malignancy - An institutional experience.

BACKGROUND: Liquid-based cytology (LBC) has been developed as an alternative for conventional cytology (CC) in cervical smears. It is now increasingly being used all over the world for cervical cancer screening. However, its role and diagnostic accuracy in bronchial wash (BW)/bronchoalveolar lavage (BAL) specimens remains undetermined. AIMS: To assess and compare the diagnostic performance and accuracy of LBC with CC for detecting malignancy in bronchial specimens. SETTINGS AND DESIGN: This was a retrospective analytical hospital-based study. MATERIALS AND METHODS: Bronchial specimens (BW/BAL) received over a period of 4.5 years were reviewed. The samples were processed by CC from June 2010 to September 2012 (2.25 years) and by LBC from October 2012 to December 2014 (2.25 years). Data were retrieved from the records of cytology laboratory and compared among both the groups. Detection rate for histologically or cytologically verified samples was calculated. RESULTS: A total of 559 cases verified by histological and cytological follow-up were evaluated. These included 247 CC cases and 312 LBC cases. The positive diagnostic rate for malignancy in CC was 28.6% whereas that for LBC was 32.9%. The negative diagnostic rates were 66.5% and 66.3% for CC and LBC, respectively. However, unsatisfactory rates had shown a good reduction from 4.4% in CC to 0.6% after LBC introduction. The smears showed more homogeneous distribution of cells with elimination of obscuring factors such as blood, inflammation, and mucus. CONCLUSIONS: The diagnostic accuracy of LBC was slightly better than CC. The unsatisfactory rates showed reduction in LBC preparation. Thus, LBC is a viable alternative to CC and has the advantages of standardization of preparation with decrease in unsatisfactory rates.

Bronchial wash cytology: A study on morphology and morphometry.

BACKGROUND: Bronchial wash cytology of lung lesions is a non/minimally invasive procedure utilized for diagnosis of pulmonary lesions. AIM: The aim of this study was to evaluate the efficacy of bronchial wash cytology in the diagnosis of bronchopulmonary lesions and assess the role of morphometry in categorizing dysplastic/malignant lesions. MATERIALS AND METHODS: All cases of bronchial wash cytology received from January 2006 to June 2010 were retrieved and reviewed. Cases with adequate clinical data or a subsequent biopsy were selected for the study and cytodiagnosis was correlated with available clinical details. Morphometry was done on alcohol fixed hematoxylin and eosin stained cytosmears using computer assisted Image Pro software. RESULTS: One hundred and seventy-six cases of the 373 cases of bronchial cytology received were included for the study. Bronchial wash cytology technique showed high specificity. Cytohistopathology correlation showed 62.06% concordance rate. Cells from normal epithelium, reactive atypia, neoplastic atypia, squamous metaplasia, non-small cell and small cell carcinoma showed a mean nuclear diameter of 7.4 µm, 11.7 µm, 13.9 µm, 13.0 µm, 10.7 µm, and 17.7 µm, respectively, which was statistically significant with P < 0.05. Multiple comparisons between various groups using analysis of variance and Bonferroni tests also showed remarkable statistical significance. CONCLUSIONS: Bronchial wash cytology has low sensitivity in detecting pulmonary lesions. It can be of value in patients with contraindication for biopsy. Morphometry can be a useful adjunct to cytomorphology, especially in situations where biopsy is contraindicated.

Accuracy of bronchial brush and wash specimens prepared by the ThinPrep method in the diagnosis of pulmonary small cell carcinoma.

BACKGROUND: ThinPrep bronchial brush and wash accuracy in the diagnosis of pulmonary small cell carcinoma (pSCCa) and measured as sensitivity, specificity, positive and negative predictive values (PPV and NPV) is incompletely studied or unknown. METHODS: Specimens collected over 5 years from 199 pSCCa and 938 negative (Neg) for pulmonary cancer individuals were selected by linking the laboratory file with the cancer registry. Results other than unsatisfactory were classified as true-positive and -negative, and false-positive and -negative tests so as to calculate accuracy estimates. Slides of all false-negative and -positive and randomly selected samples of true-positive and -negative tests were evaluated for 11 abnormal cell features typical of pSCCa in conventional preparations: distribution differences by disease status were tested for significance. RESULTS: There were 129 brush and 170 wash in the pSCCa group and 365 brush and 1153 wash in the Neg group. Of all specimens, 1.2% were unsatisfactory. Brush sensitivity, specificity, PPV, and NPV were 61.9%, 99.4%, 97.5%, and 88%, respectively. Wash frequencies were 53.3%, 98.8%, 86.5%, and 93.5%, respectively. Abnormal cell features occurred in 29.9% of the selected pSCCa and 4.7% of the Neg specimens, and distribution differences were significant for each feature (P < .001). CONCLUSIONS: Unsatisfactory brush and wash specimens are infrequent in the diagnosis of pSCCa, and both have moderate sensitivity and high specificity, PPV, and NPV. pSCCa abnormal cell features resemble those seen in conventional preparations and can distinguish specimens with pSCCa from those negative for pulmonary cancer.

Comparison between cytologic examination of fungi in bronchial washings and bronchoalveolar lavage specimens and culture: a review of 100 cases with emphasis on diagnostic pitfalls.

INTRODUCTION: Microbiology culture is the "gold standard" for diagnosis of fungal infections; however, culture has a lengthy turnaround time. A more timely assessment is possible with cytology and Gomori-methamine silver (GMS) staining. MATERIALS AND METHODS: A total of 100 respiratory tract specimens with a positive fungal Gomori-methamine silver stain and corresponding culture were selected. The cytology slides were reviewed for factors contributing to discrepant results. Specimens were classified as 2 types of variances: interpretative and sampling. Concordant diagnoses were also evaluated. RESULTS: Eighty-two cases had fungal organisms that grew in culture. The remaining 18 cases were composed solely of fungal organisms that did not grow in culture (17 cases with Pneumocystis jirovecii; 1 case with Pityrosporum ovale). These 18 cases were excluded from the variance analysis. Thirty-three of 82 cases (40%) had concordant cytology and microbiology results, whereas 49 cases were discrepant. Variances were both sampling (41 cases) and interpretive (8 cases). Interpretive variances were predominantly Aspergillus species misinterpreted as Candida. Difficulty identifying true septate hyphae was the major contributing factor for misinterpretation. CONCLUSIONS: Cytologic evaluation of respiratory specimens remains a useful preemptive diagnostic tool in the rapid diagnosis of fungal infection. Cytology samples significantly contribute to the diagnosis of respiratory fungi. However, interpretive variances between Aspergillus and Candida organisms are common. Awareness of the characteristic features that distinguish fungal organisms can further improve the diagnostic utility of cytology.

Detection of lung cancer using MAGE A1-6 and SSX4 RT-PCR expression profiles in the bronchial wash fluid.

PURPOSE: Bronchial wash fluid may be a useful for detecting lung cancer. To increase the detection rates, we performed molecular analysis with using MAGE A1-6 and SSX4 RT-PCR on bronchial wash fluid specimens. MATERIALS AND METHODS: We obtained 57 lung cancer tissue specimens by bronchoscopic biopsy and 131 bronchial washes from 96 patients with lung cancer and 35 patients with benign lung diseases. The MAGE A1-6 and SSX4 gene expressions were investigated in the cancer tissue specimens and bronchial wash fluids. We evaluated the positive detection rates of these methods according to the cytology results and the clinical findings. RESULTS: For the cancer tissue specimens and the bronchial wash fluid, the positive detection rate of MAGE or SSX4 was 91.2% and 75.0%, respectively. Combined MAGE and SSX4 PCR and cytology tests showed an 83.3% detection rate for the bronchial wash fluid. From bronchial washes of patients with benign lung diseases, the positive rates of using MAGE or SSX4 was 11.4%. In the bronchial wash fluid of lung cancer patients, 66.7% of the peripheral cancers were detected by MAGE or SSX4, while examination with cytology did not detect any peripheral lung cancer. CONCLUSION: The application of both MAGE and SSX4 showed high sensitivity and specificity for the detection of lung cancer. Thus, MAGE and SSX4 RT-PCR may be effectively utilized as additional methods to improve detection of lung cancer with using bronchial wash fluids.