Bronchial Washing Fluid Research Articles

Coronavirus: A Laboratory Perspective

COVID-19 is a virus of the species of the Family coronaviridae known as as SARS-COV-2. This virus is easily contracted and/transmitted from an infected person to another healthy individual and has continued to spread rapidly. The aim of this review is to identify laboratory methods used in the diagnosis of COVID-19 infection. COVID-19 test are aimed at detecting active infection, or past infection, or an immunization so as to treat and curb the further spread of the virus. The initial viral detection is typically carried out with the upper respiratory tract (URT) sample. Repeated testing is particularly helpful and essential if a patient has a clinical appearance of viral pneumonia, radiographic results consistent with pneumonia and/or a history of potential exposure. The Centre of Disease Control and Prevention, CDC recommends the collection of specimens from the lower respiratory tract, upper respiratory tract, and the blood. The lower respiratory tract sample includes; the sputum, broncheoalveolar lavage, bronchial wash, tracheal aspirate, and pleural fluid. The upper respiratory tract specimens include; the nasopharyngeal swab, and oropharyngeal swab (NP/OP swabs). Some laboratory techniques developed and in use for the detection of Covid-19 are; nucleic acid amplification tests (NAATs), antibody detection, and viral antigen detection. The role of the laboratory assay in diagnosis of COVID-19 infection or disease cannot be under-estimated, timing and site of specimen collected must be followed by adequate professional training to ensure result accuracy. This review provides information on available laboratory techniques for the diagnosis of the viral infection and their potential merits and limitations.

Prevalence of Antibiotics Resistance in the Isolated Bacteria from Bronchial Washing Fluids in Ramadi Teaching Hospital, Iraq

Prevalence of Antibiotics Resistance in the Isolated Bacteria from Bronchial Washing Fluids in Ramadi Teaching Hospital, Iraq

Identifying Genomic Alterations in Small Cell Lung Cancer Using the Liquid Biopsy of Bronchial Washing Fluid.

Objective: With the rapid development of cancer genomics and immunomics, some new treatments of small cell lung cancer (SCLC) are emerging. However, there are limitations to the clinical use of tumor tissue. Our study aimed to evaluate the potential use of bronchial washing fluid (BWF) in the liquid biopsy of SCLC.Methods: Twenty-one extensive SCLC (ES-SCLC) patients were enrolled in this study. For all patients, four sample types, BWF supernatant (BWFs), BWF precipitate (BWFp), plasma and tumor tissue, were collected before receiving chemotherapy, and one type, plasma, was collected after chemotherapy. All samples were conducted to NGS using the 1021-gene panel. The concordance rates of genomic profiling using NGS in the four types of samples were evaluated. Multiple clinical information was analyzed for correlation.Results: We successfully tested 20 BWFs samples, 21 BWFp samples, 21 tumor tissue samples, 20 pre-treatment plasma, and 13 post-treatment plasma of these 21 patients. The detectability of somatic mutations was 100% for BWFs, BWFp, tumor tissues, and post-treatment plasma, and only one pre-treatment plasma was absent with any mutation. Matched tumor tissue, BWFs, BWFp, and pre-treatment plasma samples were subsistent for 19 patients. For these patients, 204 genomic alterations were identified in tissue samples, while 189 (92.6%), 175 (85.5%), and 163 (79.9%) alterations were detected in the matched BWFs, BWFp, and pre-treatment plasma, respectively. Moreover, we found that the three tumor markers associated with SCLC have a lower sensitivity than genomic alterations. The endocrine resistance pathway was found enriched in hyponatremia patients which may be related to the hyponatremia. The TMBs of BWF, BWFp, and pre-treatment plasma samples all had a strong correlation with that of tissue samples. Both the VAF and the MVAF of mutations in post-treatment plasma were less than those in pre-treatment plasma, which was in accordance with the evaluation of curative effect.Conclusions: For ES-SCLC patients, the liquid biopsy of BWF showed a highly potential advantage to identify DNA alterations, which suggested that genomic analysis of BWF liquid biopsy may have clinical value as a supplement for tissue and blood detection. Through the restricted validation, it can be widely used in routine clinical practice.

Performance of the Xpert® MTB/RIF assay in the rapid diagnosis of tracheobronchial tuberculosis using bronchial washing fluid.

ObjectiveTo assess the diagnostic value of the Xpert® MTB/RIF (GeneXpert) assay for tracheobronchial tuberculosis (TBTB) using bronchial washing fluid (BWF).MethodsThis retrospective study enrolled patients suspected of having TBTB and patients with non-TB pulmonary disease as controls. BWF were used to undertake acid-fast bacillus (AFB) smears, the GeneXpert assay and the Lӧwenstein–Jensen (LJ) culture method. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were compared among BWF AFB smears, BWF GeneXpert and the BWF LJ culture method.ResultsA total of 130 patients with TBTB and 102 patients with non-TB pulmonary disease were enrolled in the study. Sputum AFB smears were positive in 62 of 130 patients (47.7%) with TBTB. Using the clinical diagnosis of TBTB as the gold standard, the sensitivity, specificity, PPV and NPV of the three methods using BWF were as follows: 93.1%, 99.0%, 99.2% and 91.8% for BWF GeneXpert; 73.1%, 100.0%, 100.0% and 74.5% for BWF LJ cultures; 53.8%, 99.0%, 98.6% and 62.7% for BWF AFB smears. The diagnostic yield of BWF GeneXpert was significantly higher compared with BWF cultures for type III and IV TBTB.ConclusionThe Xpert® MTB/RIF assay using BWF exhibited higher sensitivity than bacteriological diagnostic methods and was particularly useful for the early diagnosis of smear-negative TBTB.

Usefulness of Xpert MTB/RIF Ultra to Rapidly Diagnose Sputum Smear-Negative Pulmonary Tuberculosis Using Bronchial Washing Fluid.

This study evaluated the performance of the Xpert MTB/RIF Ultra assay (Xpert Ultra) to detect smear-negative pulmonary tuberculosis (PTB). Xpert Ultra assay was prospectively performed using bronchial washing fluid (BWF) in comparison to COBAS TaqMan MTB (COBAS) assay and mycobacterial culture. Of the 165 enrolled participants, 27 (16.4%) had PTB based on composite reference standard and 16 (9.7%) had culture-confirmed PTB. By the composite reference standard of PTB, the sensitivity of Xpert Ultra (63.0, 95% confidence interval, CI, 42.4–80.6%) was higher than the COBAS assay (25.9%, P = 0.006), BWF-culture (33.3%, P = 0.029) and sputum-culture (37.0%, P = 0.057). Meanwhile, the specificity of Xpert Ultra was 99.3% which was slightly lower than the 100.0% specificity of the COBAS assay (P = 1.000) and cultures (P = 1.000). Against the reference standard of culture-confirmed PTB, Xpert Ultra also had a higher sensitivity (62.5, 95% CI, 35.4–84.8%) than the COBAS assay (31.3%, P = 0.077) and was similar to BWF-culture (56.3%, P = 0.719) and sputum-culture (62.5%, P = 1.000). However, one subject with previously treated old PTB had a false-positive result on the Xpert Ultra assay. This prospective study showed Xpert Ultra assay using BWF had better sensitivity than COBAS assay and mycobacterial cultures but could represent a false positive in patients with inactive old PTB.

Compared to plasma, bronchial washing fluid shows higher diagnostic yields for detecting EGFR-TKI sensitizing mutations by ddPCR in lung cancer

BackgroundThe rate of diagnosis of advanced lung adenocarcinoma must be improved. In this study, we compared the detection rates of EGFR-tyrosine kinase inhibitor-sensitizing mutations (mEGFRs) in bronchial washing fluid (BWF) and the plasma of patients with lung adenocarcinoma using the tissue genotype as the standard reference.MethodsPaired blood and BWF specimens were collected from 73 patients with lung cancer. The tumor EGFR mutation status was determined by genotyping of the plasma and BWF samples using droplet digital PCR (ddPCR).ResultsThe study cohort included 26, 10, 10, and 27 patients with stage I, II, III, and IV disease. Of the 73 cases, 35 had a wild-type EGFR, and 19 had the L858R substitution and exon 19 deletion mutations. The areas under the receiver operator characteristic curves for sensitivity vs. specificity of ddPCR were 0.895 [95% confidence interval (CI): 0.822–0.969] for BWF and 0.686 (95% CI: 0.592–0.780) for plasma (p < 0.001). The fractional abundance was higher in BWF of the mEGFR-positive cases than in the plasma (p = 0.004), facilitating easy threshold setting and discrimination between mEGFR-positive and negative cases. When genotyping results obtained using plasma and BWF were compared for early lung cancer (stages I–IIIA), the diagnostic yields were significantly higher for BWF ddPCR, and the same tendency was observed for the advanced stages, suggesting that the BWF data may reflect the genotype status in early-stage patients.ConclusionsThe mEGFR genotyping results obtained using BWF showed a higher diagnostic efficacy than did those obtained using the plasma. Thus, BWF-based genotyping may be a useful substitute for that using plasma in lung cancer.

Identifying genomic alterations in small cell lung cancer using the liquid biopsy of bronchial washing fluid.

e21098 Background: With the rapid development of cancer genomics, the precision medicine of small cell lung cancer (SCLC) is emerging. However, there are limitations to the clinical use of tumor tissue and peripheral blood biopsies. The main purpose of this study was to evaluate the potential use of bronchial washing fluid (BWF) in the liquid biopsy of SCLC. Methods: Twenty-one SCLC patients diagnosed in 2019 were enrolled in this study. BWF (separated as supernatant and precipitate), treatment-naive plasma and tumor tissue samples were collected from all of patients and subjected to next-generation sequencing (NGS) using a 1021-gene panel. The concordance rates of genomic profiling using NGS in these four types of samples were evaluated. Results: Of these 21 patients, 20 BWF supernatant (BWFs) samples, 21 BWF precipitate (BWFp) samples, 21 tumor tissue samples and 20 plasma samples were successfully tested. The detectability of somatic mutations was 100% for BWFs, BWFp and tumor tissues, and only one plasma was absent with any mutation. Matched tumor tissue, BWFs, BWFp and plasma samples were subsistent for 19 patients. For these patients, 204 genomic alterations were identified in tissue samples, of which 189 (92.6%), 175 (85.5%) and 163 (79.9%) alterations were detected in the matched BWFs, BWFp and plasma samples, respectively. Moreover, tumor mutation burden (TMB) was also calculated. Compared with the proportion of TMB-H samples in tissue samples counting 61.9% (13/21), 60% (12/20) of BWFs samples and 52.38 % (11/21) of BWFp samples were TMB-H (defined as more than or equal to 9 mutations per megabase), which was a molecular biomarker that can be used in immunotherapy efficacy prediction. The TMBs of BWFs, BWFp and treatment-naive plasma samples all had strong correlation with that of tissue samples. The TMB of BWFs had the strongest correlation (Pearson r = 0.9512, p &lt; 0.0001), and the TMB of treatment-naive plasma had relatively lower correlation (Pearson r = 0.8782, p &lt; 0.0001) compared with those of BWFs (Pearson r = 0.936, p &lt; 0.0001) and BWFp (Pearson r = 0.8782, p &lt; 0.0001). Conclusions: For SCLC patients, the liquid biopsy of BWF showed high potential to identify DNA alterations and calculate TMB grades, which suggested that genomic analysis of BWF liquid biopsy may have clinical value in predicting the effectiveness of targeted therapy and immunotherapy. It can be widely used in routine clinical practice.

Assessment of Donor Lung Aspiration on the Ex Vivo Lung Perfusion (EVLP) Platform

Donor aspiration of gastric contents represents a significant risk for lung injury and severe primary graft dysfunction after lung transplantation. Concern about possible aspiration is a common reason for clinicians to decline donor lungs for transplantation. The presence of bile acid in donor bronchial wash (BW) or bronchoalveolar lavage (BAL) fluid may indicate that a donor has aspirated. Previous reports have also shown that aspiration of bile acids cause increased inflammation in the lung. Ex vivo lung perfusion (EVLP) has become an important platform to assess questionable or "marginal" donors. In this study, we examined the relationship between possible donor aspiration and physiologic and biologic assessment of inflammation on EVLP. Large airway BW samples were collected from 270 donors (n=111 non-EVLP, n=159 EVLP) prior to transplantation. Bile acid levels were measured in BW samples using a commercially available colorimetric assay and clinically approved platform. Protein and biochemical measurements were made in perfusate samples collected hourly after the start of EVLP. The majority of donor lungs with clinically suspected aspiration were successfully transplanted after EVLP assessment (58% [23/40]). Donor bile acid levels were significantly higher in donor lungs that were ultimately declined for transplantation at the end of EVLP (0.28 vs. 0.12 μM, p<0.05, n=143). During EVLP there was a positive correlation between bile acid in the donor BW and inflammation (IL-8 levels (r=0.19, p<0.05, n=159)). Similarly, there was an increase in biochemical markers of cell death ([Ca2+]) in the EVLP perfusate of donors with high bile acid levels in BW (r=0.27, p<0.001, n=142). Following EVLP and transplantation, we observed no significant effects of donor bile acid levels on recipient ICU length-of-stay (r=0.09, p=0.35, n=117). Suspected donor aspiration can be assessed using the EVLP platform. Bile acid levels measured in the donor lung are associated with increased inflammation and lung injury observed during EVLP, but not recipient outcomes post-EVLP. Therefore, EVLP may be an ideal platform to identify marginal donor lungs suspected of aspiration but are ultimately suitable for clinical transplantation.

Clinical Characteristics and Outcomes of Surgically Resected Solitary Pulmonary Nodules Due to Nontuberculous Mycobacterial Infections.

Background: Limited data are available regarding the detailed characteristics and outcomes of surgically resected nontuberculous mycobacterial (NTM) granulomas. Methods: We evaluated the characteristics of 49 NTM granulomas presenting as solitary pulmonary nodules (SPNs) between January 2007 and December 2016. Results: Twenty-five patients (51%) were male and 27 (55%) were never-smokers. Seven (14%) patients had a history of tuberculosis. More than half (51%) of patients were asymptomatic. On chest computed tomography, the median SPN diameter was 18 mm, and approximately half of all SPNs (49%) were located in the upper lobes on chest computed tomography. NTM strain were preoperatively isolated from sputum (46%, 12/26), bronchial wash fluid (54%, 14/26), and needle biopsy specimens (12%, 3/26). Mycobacterium avium (71%, 22/31) was the organism most commonly isolated, followed by Mycobacterium intracellulare (16%, 5/31). Postoperative pneumothorax and atelectasis developed in four (8%) patients and one (2%) patient, respectively. Five patients received postoperative antibiotic therapy. Over a median follow-up period of 18.0 months, one patient with residual lesions after surgery started macrolide-based therapy due to aggravated symptoms. Conclusions: Most NTM granulomas can be treated completely by surgical resection without antibiotic therapy, and microbiological examination of surgical specimens is important for optimal management.

DISCREPANCIES IN BRONCHIAL WASH AND LAVAGE: THE DUBIOUS CASE OF VARICELLA PNEUMONIA

SESSION TITLE: Monday Fellow Case Report Posters SESSION TYPE: Fellow Case Report Posters PRESENTED ON: 10/21/2019 02:30 PM - 03:15 PM INTRODUCTION: Varicella pneumonia is a rare but serious infection caused by Varicella Zoster virus (VZV). Diagnosis is attained by clinical presentation, radiologic findings & is aided by detection of VZV in respiratory secretions by polymerase chain reaction (PCR). We present a diagnostic dilemma when two tests have a discrepancy. CASE PRESENTATION: A 41 year old female presented with left upper quadrant (LUQ) abdominal pain, hypotension, melena & somnolence which prompted intubation. She had medical history of end stage liver disease due to alcoholic cirrhosis, esophageal varices, refractory ascites & encephalopathy. Endoscopy showed varices & small bowel ulcer; & paracentesis ruled out bacterial peritonitis. After treatment with octreotide, nexium & prophylactic ceftriaxone; she was was successfully extubated. 3 days later, she developed a new, vesicular, dermatomal rash of shingles on LUQ. This was followed by new extensive multifocal infiltrates, crazy paving pattern & diffuse ground glass opacities on chest imaging (Figure) along with hypoxic respiratory failure which warranted re-intubation. A bronchoscopy was done which showed thick white secretions on bronchial washing (BW) & cloudy fluid on bronchoalveolar lavage (BAL) of right middle lobe & left upper-lingular segment. VZV PCR resulted positive on BW but negative on BAL. Patient was started on Varicella-Zoster immune globulin (VZIG) and acyclovir based on clinical presentation and radiologic features which led to clinical improvement. DISCUSSION: Varicella pneumonia can manifest 1-6 days after onset of the typical rash & is highly infectious with a secondary attack rate of > 90%. In the presence of clinical features of VZV infection, the diagnosis is mediated by chest imaging showing nodular or interstitial pneumonitis with ground glass opacities. Molecular diagnostic assays using PCR are often employed and have sensitivity as high as 96% for detection of VZV (1). However, it can be limited by false positive rate due to contamination and depends on the methodology (2) as well as specimen source although BAL has high sensitivity (3). Our patient developed respiratory failure with radiologic findings shortly after typical zoster rash. Despite adequate sampling, there was a discrepancy in the result obtained from BAL and BW. A comparison of these two hasn’t been studied yet in respect to VZV as well as other common pathogens. We favored to treat the patient due to detection of VZV on BW & the clinical presentation. VZIG administration within 72 hours of exposure is highly efficacious for disease prevention in immunocompromised along with intravenous acyclovir for 7-10 days and has led to overall decline in mortality. CONCLUSIONS: Although VZV DNA detection by PCR is helpful in diagnosing VZV pneumonia, there is no consensus data on its utility on BW in comparison to BAL. The paucity of VZV pneumonia is a hindrance for more extensive studies. Reference #1: Mirouse A et al. Severe varicella-zoster virus pneumonia: a multicenter cohort study. Critical Care. 2017 Dec;21(1):137 Reference #2: Schmutzhard J, Riedel HM, Wirgart BZ, Grillner L. Detection of herpes simplex virus type 1, herpes simplex virus type 2 and varicella-zoster virus in skin lesions. Comparison of real-time PCR, nested PCR and virus isolation. Journal of clinical virology. 2004 Feb 1;29(2):120-6 Reference #3: Stránská R, Schuurman R, de Vos M, van Loon AM. Routine use of a highly automated and internally controlled real-time PCR assay for the diagnosis of herpes simplex and varicella-zoster virus infections. Journal of Clinical Virology. 2004 May 1;30(1):39-44 DISCLOSURES: No relevant relationships by Enambir Josan, source=Web Response No relevant relationships by Supriya Singh, source=Web Response No relevant relationships by Yasir Tarabichi, source=Web Response

Identifying actionable somatic mutations in lung cancer using cell-free DNA from bronchial washing fluid.

8528 Background: Bronchial washing is the most common technique for sampling the components of the alveolar space. Here, we evaluated the potential use of bronchial washing fluid (BWF) in liquid biopsy in lung cancer. Methods: This study enrolled 65 lung cancer patients. BWF (separated supernatant and precipitate) samples, peripheral blood lymphocytes (PBL) and formalin-fixed paraffin-embedded tissues were obtained and subjected to next-generation sequencing using a 1021-gene panel. Results: Mutations were identified in 58 (89.2%) of BWF precipitate (BWFp) samples and 64 (98.5%) of BWF supernatant (BWFs) samples, comparing with 61 (93.8%) of tumor tissues. In total, 461 somatic mutations were identified in tissues, of which 331 (71.8%) and 381 (82.6%) were detected in the matched BWFp and BWFs samples. In addition, there were 44.6% of patients carrying actionable variants identified in tissue DNA, including EGFR, ALK, ROS1, RET, etc. (Table). Similarly, there were 40.0% of BWFp samples and 44.6% of BWFs samples identified actionable variants. Moreover, tumor mutation burden (TMB) was also calculated. Nearly 9% of BWFp samples and 23% of BWFs samples were TMB-H (more than 9 mutations per megabase), comparing with 20% of tissues. Significantly, the combined results of three types of samples showed that, 49.2% of patients carrying actionable variants and 24.6% of patients with TMB-H, which suggested more patients benefit from targeted therapy or immunotherapy. Conclusions: In summary, liquid biopsy using BWF showed high potential to identify actionable mutations and to calculate TMB grade of patients with lung cancer, which might be implemented and standardized into clinical use. [Table: see text]

Prevalence of blaKPC and its occurrence with other beta-lactamases in Klebsiella pneumoniae.

BACKGROUND:Klebsiella pneumoniae (K. pneumoniae) is an important nosocomial pathogen, and the emergence of multidrug resistance in these organisms limits the treatment options for serious infections caused by them. K. pneumoniae carbapenemase (KPC) is one of the clinically significant Class A beta-lactamases.AIM AND OBJECTIVE:This study was aimed to detect the KPC and its coexistence with other beta-lactamases in K. pneumoniae.MATERIALS AND METHODS:A total of 370 isolates, collected over a period of 1 year, were included in this study. The source of these isolates were urine (n = 170), exudative specimens (n = 132), respiratory secretions such as bronchial wash, endotracheal aspirate, and pleural fluid (n = 38), and blood (n = 30). For all the isolates, antibiotic susceptibility tests by disc diffusion, modified Hodge test, and KPC screening test were done. Polymerase chain reaction (PCR) was performed for the detection of KPC and the copresence of other beta-lactamases genes.RESULTS:Among the 370 isolates, 41 were resistant to the carbapenem by disc diffusion and minimum inhibitory concentration tests. Screen test using ertapenem and the boronic acid disk was positive in 14 isolates. Only one isolate harbored KPC gene by PCR, and it was co-produced with SHV-12 and CTX-M-15.CONCLUSION:PCR remains the gold standard for detection of KPC compared with any other phenotypic methods. Early detection of these genes helps in initiating proper antibiotic treatment.

PCDHGA12 methylation biomarker in bronchial washing specimens as an adjunctive diagnostic tool to bronchoscopy in lung cancer.

The use of bronchoscopy is central to the diagnosis of lung cancer. However, the sensitivity of bronchoscopy is low. In addition, bronchial washing cytology, which is a routine adjunctive test, does not significantly improve the performance of bronchoscopy owing to its low sensitivity. To enhance the diagnostic performance of bronchoscopy, the protocadherin GA12 (PCDHGA12) methylation biomarker in bronchial washings was introduced as a novel adjunctive diagnostic test. A total of 98 patients who underwent bronchoscopy owing to suspicion of lung cancer were analyzed. Cytological examination and PCDHGA12 methylation biomarker testing of the bronchial washing fluid were performed. The performance of the tests was analyzed. The final diagnosis in 60 patients was lung cancer and in 38 patients was benign disease. The PCDHGA12 methylation biomarker had a sensitivity of 75.0%, a specificity of 78.9% and a positive predictive value (PPV) of 84.9%, whereas cytological assessment had a sensitivity of 45.0%, a specificity of 92.1% and a PPV of 90%. Patients with positive PCDHGA12 methylation test had an odds ratio for lung cancer of 11.25 (confidence interval, 4.25-29.8) compared with negative subjects. The combination of the two tests exhibited an increased sensitivity (83.3%), a specificity of 71.1% and a PPV of 82.0%. Furthermore, considering the non-diagnostic bronchoscopy group alone, the test demonstrated a sensitivity of 61.9% and a specificity of 78.9%. The results of the present study demonstrated that PCDHGA12 methylation, as a lung cancer biomarker in bronchial washings, may be a used as an adjunctive test to bronchoscopy.

P3.01-004 The Underestimated Role of Bronchial Washing Fluid in the Detection of EGFR Mutation from Lung Cancer Patients

P3.01-004 The Underestimated Role of Bronchial Washing Fluid in the Detection of EGFR Mutation from Lung Cancer Patients

Epidemiological and Clinical Characteristics of Streptococcus pneumoniae Infections in a Tertiary Care Center in Bahrain (2010-2014)

Objective: &#x0D; To describe the epidemiology of invasive and non-invasive pneumococcal disease during a 5-year period in a major hospital in Bahrain.&#x0D; &#x0D; Design and Setting: &#x0D; Retrospective study of cases of pneumococcal diseases among patient ≥ 14 years old during a five-year period (2010-2014), who attended Salmaniya Medical Complex, Bahrain.&#x0D; &#x0D; Patients and Methods:&#x0D; Laboratory data was obtained on Streptococcus pneumoniae isolates. Patients ≥14 years old of age, both inpatient and outpatient with cultures positive for Streptococcus pneumoniae were included. Only samples from blood, cerebrospinal fluid (CSF), pleural fluid, sputum, deep tracheal aspirate (DTA), bronchial wash, eye discharge and middle ear fluid were included. Demographics and clinical records from medical files were then reviewed.&#x0D; &#x0D; Results:&#x0D; A total of 130 culture samples met the inclusion criteria and were analyzed. One-hundred twenty-six patients with pneumococcal disease were identified; with 65% being non-invasive and 35% being invasive pneumococcal disease. Overall, non-bacteremic Pneumonia was the most frequent manifestation, followed by bacteremia. Pneumococcal disease was more common among male patients (62.7%). Mean age was 57.67 (15-95 years). The most common comorbidities were hypertension, Diabetes Mellitus followed by cardiovascular disease. Twenty-seven patients received mechanical ventilation during their admission, 13 patients were admitted to the intensive care unit, and 6 patients developed para-pneumonic effusion as a complication. In-hospital Mortality rate was 13.5%. &#x0D; &#x0D; Conclusion: &#x0D; This study supports the current literature where pneumococcal infection is more prevalent in elderly and those with comorbidities. Mortality was related to age, certain chronic medical illness, admission to intensive care unit and need for mechanical ventilation.

1290P - DNA repair gene expression in bronchial washing fluid as new molecular tool for clinical outcome decision

1290P - DNA repair gene expression in bronchial washing fluid as new molecular tool for clinical outcome decision

1334P - A comparison of Bronchial Washing Fluid (BWF) and histologic samples in the analysis of EGFR mutation in NSCLC patients

1334P - A comparison of Bronchial Washing Fluid (BWF) and histologic samples in the analysis of EGFR mutation in NSCLC patients

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.

Diagnosis of cytomegalovirus pneumonia by quantitative polymerase chain reaction using bronchial washing fluid from patients with hematologic malignancies

BackgroundThe incidence of cytomegalovirus (CMV) pneumonia is increasing in patients diagnosed with hematologic malignancies. The utility of CMV-DNA viral load measurement has not been standardized, and viral cut-off values have not been established. This study was designed to investigate the utility of CMV quantitative real-time PCR (qRT-PCR) using bronchial washing fluid.MethodsWe retrospectively reviewed the microbiologic and pathologic results of bronchial washing fluid and biopsy specimens in addition to the patients' clinical characteristics.ResultsA total of 565 CMV qRT-PCR assays were performed using bronchial washing fluid from patients with hematologic malignancies. Among them, 101 were positive for CMV by qRT-PCR; of these, 24 were diagnosed with CMV pneumonia and 70 with CMV infection, and 7 were excluded due to a diagnosis of invasive pulmonary aspergillosis rather than viral pneumonia. The median CMV load determined by qPCR was 1.8 × 105 copies/mL (3.6 103-1.5 × 108) in CMV pneumonia patients and 3.0 × 103 copies/mL (5.0 × 102-1.1 × 105) in those diagnosed with CMV infection (P < 0.01). Using the ROC curve, the optimal inflection points were 18,900 copies/mL (137,970 IU/mL) in post-bone marrow transplantation (BMT) patients, 316,415 copies/mL (2,309,825 IU/mL) in no-BMT patients and 28,774 copies/mL (210,054 IU/mL) in all patients.ConclusionsThe CMV titers in bronchial washing fluid determined by qRT-PCR differed significantly between patients diagnosed with CMV pneumonia and those with CMV infection. The viral cut-off values in bronchial washing fluid were suggested for the diagnosis of CMV pneumonia, which were different depending on the BMT status.

Usefulness of flexible bronchoscopy in children with suspected pulmonary tuberculosis who have difficulty in sputum expectoration

Purpose: To assess the usefulness of flexible bronchoscopy in patients with suspected pulmonary tuberculosis (PTB) who have diffi-culty in sputum expectoration. Methods: The subjects of this study were patients who were suspected of PTB and visited the Division of Pediatric Pulmonology at a tertiary hospital from April 2006 to March 2016. PTB suspects were determined by clinical symptoms, radiologic findings, and im-munologic studies. We aimed to examine the value and safety of bronchoscopy in diagnosis and differential diagnosis of PTB in PTB- suspected patients. The diagnostic criteria for PTB were defined when Mycobacterium tuberculosis was cultured in the sputum speci-men or in the bronchial washing fluid. Results: A total of 19 PTB suspects were included. One patient was diagnosed with PTB by using the sputum study. However, the re-maining 18 patients could not expectorate sputum or showed no evidence of Mycobacterium tuberculosis infection from the spu-tum study. Of the 18 patients, 15 underwent bronchoscopy. After bronchoscopy, 6 patients were diagnosed with PTB and 9 patients were diagnosed with Mycoplasma, viral, or fungal pneumonia, and tumors. For antituberculous drug resistance, there were 1 case of isoniazid (INH) resistance and 1 case of concurrent resistance to INH and prothionamide. There was no multidrug-resistant tubercu-losis. None of the patients had significant complications due to bronchoscopy. Conclusion: Flexible bronchoscopy appears to be a definitive and safe procedure for the differential diagnosis of patients suspect-ing PTB in children who have difficulty expectorating sputum. (Allergy Asthma Respir Dis 2017;5:287-293)