Bronchoalveolar Lavage Specimens Research Articles

MicroRNA Monitoring in Human Alveolar Macrophages from Patients with Smoking-Related Lung Diseases: A Preliminary Study.

Chronic obstructive pulmonary disease (COPD) is a progressive lung disease that is commonly considered to be a potent driver of non-small cell lung cancer (NSCLC) development and related mortality. A growing body of evidence supports a role of the immune system, mainly played by alveolar macrophages (AMs), in key axes regulating the development of COPD or NSCLC phenotypes in response to harmful agents. MicroRNAs (miRNAs) are small non-coding RNAs that influence most biological processes and interfere with several regulatory pathways. The purpose of this study was to assess miRNA expression patterns in patients with COPD, NSCLC, and ever- or never-smoker controls to explore their involvement in smoking-related diseases. Bronchoalveolar lavage (BAL) specimens were collected from a prospective cohort of 43 sex-matched subjects to determine the expressions of hsa-miR-223-5p, 16-5p, 20a-5p, -17-5p, 34a-5p and 106a-5p by RT-PCR. In addition, a bioinformatic analysis of miRNA target genes linked to cancer was performed. Distinct and common miRNA expression levels were identified in each pathological group, suggesting their possible role as an index of NSCLC or COPD microenvironment. Moreover, we identified miRNA targets linked to carcinogenesis using in silico analysis. In conclusion, this study identified miRNA signatures in AMs, allowing us to understand the molecular mechanisms underlying smoking-related conditions and potentially providing new insights for diagnosis or pharmacological treatment.

Detection of Feline Coronavirus in Bronchoalveolar Lavage Fluid from Cats with Atypical Lower Airway and Lung Disease: Suspicion of Virus-Associated Pneumonia or Pneumonitis.

The premortem understanding of the role of feline coronavirus (FeCoV) in the lungs of cats is limited as viruses are seldom inspected in the bronchoalveolar lavage (BAL) specimens of small animal patients. This study retrospectively analyzed the prevalence of FeCoV in BAL samples from cats with atypical lower airway and lung disease, as well as the clinical characteristics, diagnostic findings, and follow-up information. Of 1162 clinical samples submitted for FeCoV RT-nPCR, 25 were BAL fluid. After excluding 1 case with chronic aspiration, FeCoV was found in 3/24 (13%) BAL specimens, with 2 having immunofluorescence staining confirming the presence of FeCoV within the cytoplasm of alveolar macrophages. The cats with FeCoV in BAL fluid more often had pulmonary nodular lesions (66% vs. 19%, p = 0.14) and multinucleated cells on cytology (100% vs. 48%, p = 0.22) compared to the cats without, but these differences did not reach statistical significance due to the small sample size. Three cats showed an initial positive response to the corticosteroid treatment based on the clinical signs and radiological findings, but the long-term prognosis varied. The clinical suspicion of FeCoV-associated pneumonia or pneumonitis was raised since no other pathogens were found after extensive investigations. Further studies are warranted to investigate the interaction between FeCoV and lung responses in cats.

Evaluation of Alinity m CMV assay performance for detecting CMV in plasma, cerebrospinal fluid, and bronchoalveolar lavage specimens

Accurate detection and quantification of cytomegalovirus (CMV) is crucial to preventing adverse outcomes in immunocompromised individuals. Current assays were developed for use with plasma specimens, but CMV may be present in bronchoalveolar lavage (BAL) fluid and cerebrospinal fluid (CSF). We evaluated the performance of the Abbott Alinity m CMV assay compared to the Abbott RealTime CMV assay for quantification of CMV in plasma, BAL, and CSF specimens. To evaluate clinical performance, 190 plasma, 78 BAL, and 20 CSF specimens were tested with the Alinity m assay and compared to the RealTime assay. The Alinity m CMV assay showed high precision (SD <0.01 to 0.13) for all 3 specimen types. Clincal plasma and BAL specimens with quantifiable CMV DNA demonstrated strong correlation to RealTime CMV assay results (r2 = 0.9779 for plasma, r2 = 0.9373 for BAL). The Alinity m CMV assay may be useful for quantification of CMV in plasma, BAL, and CSF specimens.

Application of a multiplex molecular pneumonia panel and real-world impact on antimicrobial stewardship among patients with hospital-acquired and ventilator-associated pneumonia in intensive care units

BackgroundThe optimal timing for applying the BioFire FilmArray Pneumonia Panel (FAPP) in intensive care unit (ICU) patients with hospital-acquired pneumonia (HAP) or ventilator-associated pneumonia (VAP) remains undefined, and there are limited data on its impact on antimicrobial stewardship. MethodsThis retrospective study was conducted at a referral hospital in Taiwan from November 2019 to October 2022. Adult ICU patients with HAP/VAP who underwent FAPP testing were enrolled. Patient data, FAPP results, conventional microbiological testing results, and the real-world impact of FAPP results on antimicrobial therapy adjustments were assessed. Logistic regression was used to determine the predictive factors for bacterial detection by FAPP. ResultsAmong 592 respiratory specimens, including 564 (95.3%) endotracheal aspirate specimens, 19 (3.2%) expectorated sputum specimens and 9 (1.5%) bronchoalveolar lavage specimens, from 467 patients with HAP/VAP, FAPP testing yielded 368 (62.2%) positive results. Independent predictors for positive bacterial detection by FAPP included prolonged hospital stay (odds ratio [OR], 3.14), recent admissions (OR, 1.59), elevated C-reactive protein levels (OR, 1.85), Acute Physiology and Chronic Health Evaluation II scores (OR, 1.58), and septic shock (OR, 1.79). Approximately 50% of antimicrobial therapy for infections caused by Gram-negative bacteria and 58.4% for Gram-positive bacteria were adjusted or confirmed after obtaining FAPP results. ConclusionsThis study identified several factors predicting bacterial detection by FAPP in critically ill patients with HAP/VAP. More than 50% real-world clinical practices were adjusted or confirmed based on the FAPP results. Clinical algorithms for the use of FAPP and antimicrobial stewardship guidelines may further enhance its benefits.

Analysis of 332 fiberoptic bronchoscopies performed in a respiratory intensive care unit: a retrospective study.

Fiberoptic bronchoscopy (FOB) is widely used in the intensive care unit for diagnostic and therapeutic purposes. Our study aimed to evaluate FOB's indications, complications, and clinical outcomes in our intensive care unit's mechanically ventilated patients and identify the microorganisms grown in bronchoalveolar lavage (BAL) specimens. Between January 1, 2022, and June 30, 2023, a total of 332 FOBs were performed on 178 patients in the respiratory intensive care unit. Patients' mean age was 64±19.4 years. Females accounted for 65 (36.6%) and males accounted for 113 (63.4%) of the cases. Leading diagnoses included pneumonia (59.5%), acute respiratory distress syndrome (ARDS) (20.7%), sepsis (17.9%), chronic obstructive pulmonary disease (COPD) attack (21.9%), pulmonary embolism (10.1%), lung malignancy (43.8%), hemoptysis (8.9%), heart failure (15.1%), neurological/neuromuscular conditions (8.4%), and post cardiopulmonaryresuscitation (CPR) (2.8%). FOB purposes were BAL retrieval (43.6%), secretion clearance (30.4%), guided tracheostomy (11.7%), atelectasis (8.7%), and hemoptysis (5.4%). Hypoxemia marked the primary FOB complication (3.6%). Other issues encompassed hypotension (1.5%), bradycardia (1.2%), bleeding (1.2%), tachycardia (0.9%), and hypertension (0.6%). No statistical significance was found in arterial blood gas pH, arterial partial pressure of oxygen (PaO2), and arterial partial pressure of carbon dioxide (PaCO2) values before and after the FOB procedure (p>0.05). Predominant pathogens in aspiration samples were non-albicans Candida (28.9%), Klebsiella pneumoniae (24.8%), Pseudomonas aeruginosa (14.4%), and Acinetobacter baumannii (11.7%). FOB is an important diagnostic and therapeutic method with a low complication rate when performed by an experienced team with appropriate indication in the intensive care unit.

Spectrum of pathogens and their antimicrobial susceptibility pattern isolated from bronchoalveolar lavage specimens at a hilly area in Uttarakhand.

Lower respiratory tract infections (LRTIs) are associated with significant morbidity and mortality in all age groups, especially young children and the elderly population. Various gram-positive and gram-negative organisms such as Streptococcus spp., Pseudomonas spp., and Klebsiella spp. have been implicated as a pathogen in bronchoalveolar lavage (BAL) specimens collected from such patients. The present study is aimed at assessing the spectrum of the bacterial isolates and determining the antimicrobial resistance obtained from the BAL fluid from admitted patients at various wards and intensive care units (ICUs) of a tertiary care hospital in Dehradun. This will be the stepping stone in our efforts toward becoming a future antimicrobial steward and framing local antibiograms based on such data. Two hundred BAL specimens were collected from patients admitted to various wards and ICUs of the hospital who were suffering from LRTI. The BAL specimens were subjected to direct microscopy and culture. Identification and susceptibility testing were performed. The most predominant isolates were Pseudomonas aeruginosa (16/39 (41.02%)) followed by Klebsiella pneumoniae (7/39 (17.94%)) and Acinetobacter spp. (6/39 (15.38%)). Sixty-five percentage of Pseudomonas aeruginosa, 71% of Klebsiella pneumoniae, and 83% of Acinetobacter spp. showed intermediate results with colistin. Nonfermenters constitute a significant group of organisms isolated from bronchoalveolar lavage (BAL) specimens in patients with lower respiratory tract infections (LRTIs). Hence, it is extremely important to correctly identify and determine the resistance pattern of such isolates so that appropriate empirical therapy can be initiated for the benefit of the patient.

Diagnostic Value of Bronchoalveolar Lavage in Sputum-Smear Negative Pulmonary Tuberculosis

Introduction: Bronchoalveolar lavage has a high diagnostic utility for cytology and bacteriology. The aim of this study was to assess the diagnostic value of bronchoalveolar lavage (BAL) which underwent routine bronchoscopy for evaluation of lung disease. Materials and Methods: This is a hospital based descriptive study done from July 1, 2022, to June 30, 2023. Eighty bronchoalveolar lavage (BAL) cases were analyzed for differential count, cytological evaluation and bacteriological examination. All cases were included which were sent as BAL specimen to the laboratory department. Bronchoscopy was done as an outpatient procedure and lavage fluid obtained was analyzed. Result: Out of 80 cases, 51 were male and 29 were female. The age ranged from 10 to 80 years. The prevalence of tuberculosis in sputum smear negative cases was found to be 29%. Among 80 cases, eight cases were unsatisfactory, twenty-three cases were of tuberculosis, one case was of fungal infection, two cases were of malignancy, forty cases were of small airway infection and six cases were satisfactory but with no diagnostic value. Conclusion Bronchoalveolar lavage is valuable in diagnosis of tuberculosis, infections and malignancies.

Expression of Melanoma Antigen Genes A11 and A12 in Non-Small Cell Lung Cancer

Highlights:1. In this study, new primers designed using the semi-nested polymerase chain reaction (PCR) method were utilized to identify MAGE-A11 and MAGE-A12 expressions in specimens collected from core biopsy, forcep biopsy, and bronchoalveolar lavage.2. The histopathological analysis revealed positive expressions of MAGE-A11 and MAGE-A12 in specimens diagnosed with non-small cell lung cancer (NSCLC) as well as in specimens with no malignant cells. AbstractThe melanoma antigen gene (MAGE) belongs to the group of cancer testis antigens that are exclusively expressed in germ cells but may be re-expressed in cancer cells. The highly expressed MAGE-A subfamily in lung cancer may potentially be a diagnostic and prognostic biomarker. This study aimed to identify MAGE-A11 and MAGE-A12 expressions in lung tumors obtained from core biopsy, forceps biopsy, and bronchoalveolar lavage specimens. A cross-sectional observational study was conducted on 90 patients clinically diagnosed with lung tumors. These patients received core biopsy, forceps biopsy, and bronchoalveolar lavage interventions after ethical approval was obtained. The complementary deoxyribonucleic acid (cDNA) quality was assessed by the polymerase chain reaction (PCR) of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The assessment was performed to ascertain if all specimens exhibited positive PCR amplification of the GAPDH gene. MAGE-A11 and MAGE-A12 were identified through a semi-nested reverse transcription PCR. The positive results were detected by measuring the PCR products, with MAGE-A11 and MAGE-A12 measuring at base pairs (bp) of 858 and 496 in the first and second rounds, respectively. The expressions of MAGE-A11 and MAGE-A12 were observed in 3 (3.33%) and 40 (44.44%) out of 90 specimens, respectively. The prevalence rate of non-small cell lung cancer (NSCLC) was 31.11% (28/90). Among these cases, 3.57% (1/28) showed the expression of MAGE-A11, while 32.14% (9/28) exhibited the expression of MAGE-A12. Sixty-two (68.89%) out of 90 patients were diagnosed with no tumor cell malignancy. Out of 62 cases, 2 (3.23%) exhibited the expression of MAGE-A11, while 31 (50%) demonstrated the expression of MAGE-A12. MAGE-A11 and MAGE-A12 were detected in NSCLC and in certain specimens with a pathological diagnosis that indicated the absence of malignant cells. In conclusion, MAGE A11 and MAGE A12 have potential markers that can improve the pathological diagnosis of lung cancer. Further investigation is necessary to explore the expression of MAGE-A in correlation with lung cancer progression.

582. Comparing Broad-range PCR Testing and The Biofire® FilmArray® Pneumonia (PN) Panel in the Diagnosis of Bacterial Pneumonia

Abstract Background Given the low sensitivity of conventional microbial isolation methods for identifying respiratory pathogens in bacterial pneumonia, target-specific syndromic multiplex real-time PCR panels have been used in conjunction with culture methods to improve diagnostic yield. Additionally, broad-range polymerase chain reaction (BR-PCR) targeting bacterial 16s rRNA conserved region has shown higher sensitivity with certain specimen types, so we sought to evaluate the clinical performance of BR-PCR performed on bronchoalveolar lavage (BAL) specimens in comparison to The Biofire® FilmArray® Pneumonia (PN) Panel (BioFire Diagnostics, Salt Lake City, UT, USA). Methods A retrospective chart review was performed on all BAL specimens that had both a PN panel test and BR-PCR performed from January 2020 to May 2022 at all Indiana University affiliated hospitals. The PN panel test was performed in-house as per laboratory protocol, while BR-PCR was performed in a reference laboratory. Outcomes assessed included turn-around times (TAT), sensitivity and specificity of BR-PCR and clinical impact, if any. Results A total of 68 BAL specimens from 53 patients were identified (83% of patients were immunocompromised). Percent positivity for the PN panel was 19% and that of BR-PCR was 18%. With the PN panel used as the gold standard, the sensitivity and specificity of BR-PCR was 85% and 98%, respectively. Only one respiratory organism was detected by BR-PCR but not by the PN panel, and it was not considered pathogenic or to have a significant clinical impact. The median TAT for the PN panel was 2.1 hours (1.8, 3.2) versus 7.8 days (6.9, 10.4) for BR-PCR. Conclusion In our cohort of patients, BR-PCR testing was not superior to the Biofire® FilmArray® Pneumonia (PN) Panel when used to detect certain bacterial etiologies of pneumonia. Additionally, faster TAT for the panel test has the potential to enhance antimicrobial stewardship practices by enabling better antibiotic utilization. Adjunctive BR-PCR testing may be useful for clinical care when conventional testing is negative and patients are at risk for a variety of potential pathogens, including fungi. Disclosures All Authors: No reported disclosures

Epidemiology and pathogenicity of M. equirhinis in equine respiratory disorders

Mycoplasmas are pathogens involved in respiratory disorders of various animal hosts. In horses, Mycoplasma (M.) equirhinis is the species most frequently detected in clinical respiratory specimens, with a prevalence of 12–16%, but its clinical implication in equine respiratory disorders remains unclear. Here we screened 1948 clinical specimens for the presence of M. equirhinis. The samples were both tracheal washes (TW) and bronchoalveolar lavages (BAL) collected by veterinarians in France in day-to-day work between 2020 and 2022. The samples were associated with a standardized form that served to collect key general and clinical information, such as horse age, breed, and living environment. M. equirhinis was detected using a combination of culture and post-enrichment PCR. Other diagnostic data included virology and bacteriology as well as neutrophil counts, when available. Prevalence of M. equirhinis was examined as a function of a clinical score based on four significant clinical signs (nasal discharge, cough, dyspnoea, and hyperthermia). Multivariate logistic regression analysis was run to identify risk factors for the presence of M. equirhinis, and comparative prevalence analysis was used to test for association with other bacteria and viruses. TW and BAL were analysed independently, as we found that TW samples were associated with a higher prevalence of M. equirhinis. As prevalence remained steady whatever the clinical score, M. equirhinis cannot be considered a primary pathogen. M. equirhinis was more frequently isolated in thoroughbreds and trotters and in horses living exclusively stabled compared to other horses or other living environments. M. equirhinis was never detected in BAL specimens with a ‘normal’ neutrophil count, i.e. 5%, suggesting it could be associated with an inflammatory response, similar to that observed in equine asthma. Prevalence of M. equirhinis was shown to increase in the presence of other bacteria such as Streptococcus equi subsp. zooepidemicus (S. zoo) or viruses, and S. zoo load was higher in M. equirhinis-positive samples, suggesting a potential increase of clinical signs in the event of co-infection.

The transcriptional and phenotypic characteristics that define alveolar macrophage subsets in acute hypoxemic respiratory failure

The transcriptional and phenotypic characteristics that define alveolar monocyte and macrophage subsets in acute hypoxemic respiratory failure (AHRF) are poorly understood. Here, we apply CITE-seq (single-cell RNA-sequencing and cell-surface protein quantification) to bronchoalveolar lavage and blood specimens longitudinally collected from participants with AHRF to identify alveolar myeloid subsets, and then validate their identity in an external cohort using flow cytometry. We identify alveolar myeloid subsets with transcriptional profiles that differ from other lung diseases as well as several subsets with similar transcriptional profiles as reported in healthy participants (Metallothionein) or patients with COVID-19 (CD163/LGMN). We use information from CITE-seq to determine cell-surface proteins that distinguish transcriptional subsets (CD14, CD163, CD123, CD71, CD48, CD86 and CD44). In the external cohort, we find a higher proportion of CD163/LGMN alveolar macrophages are associated with mortality in AHRF. We report a parsimonious set of cell-surface proteins that distinguish alveolar myeloid subsets using scalable approaches that can be applied to clinical cohorts.

Lophomonas Infection in Patients with Respiratory Diseases in Southeastern Iran Using Wet Mount, Giemsa and Trichrome Staining

Background: In developing countries, pulmonary infections are one of the major causes of death because the incidence and prevalence of pulmonary diseases have increased dramatically. Several species of protozoa can be found in the respiratory system. Pulmonary protozoan infections are increasingly identified in clinical settings. Protozoans within the genus Lophomonas are endo-commensals of the hindgut of arthropods. Recently the trophozoite forms of Lophomonas have been observed in human tissues. Little is known about the occurrence of these protozoa in the Iranian population. Objective: This study was designed to determine the prevalence of Lophomonas in bronchoalveolar lavage specimens from the patients with respiratory diseases referred in Kerman province, southeast of Iran. Methods: A total of 200 patients were selected by simple random sampling. BAL samples were transferred to the Parasitology lab, direct smears were prepared for each specimen and two staining methods, Giemsa and Trichrome were performed for Lophomonas microscopical identification. The data were analyzed using descriptive statistics and Chi-square test. Results: Lophomonas trophozoites were found in 48 (24%) patients, at least in one of the methods. The mean age of the patients was 58.3 years (58.1 in men, 58.4 in women). Out of 200 samples, 45 (22.5%), 30 (15%), and 11(5.5%) were positive by wet mount microscopy, Giemsa, and Trichrome staining, respectively. Conclusion: This study presented the first finding of Lophomonas infection in patients with pulmo-nary symptoms in southeast Iran.

Assessment of an automated Cytomegalovirus nucleic acid amplification test using clinical plasma, bronchoalveolar lavage, and tissue specimens.

Cytomegalovirus (CMV) causes significant morbidity and mortality in immunocompromised patients, particularly transplant recipients. Quantitation of CMV DNA in peripheral blood is used to monitor prophylactic and pre-emptive approaches to prevent CMV disease, whereas CMV DNA testing of non-plasma specimens may aid in the diagnosis of end-organ disease. The analytical performance of the FDA-approved Aptima CMV Quant Assay was evaluated using reference CMV (SeraCare) diluted in defibrinated human plasma, as well as negative bronchoalveolar lavage fluid and tissue. Agreement was determined using 100 clinical acid-citrate-dextrose (ACD) plasma specimens, 77 bronchoalveolar lavage (BAL) fluids, and 101 tissues previously tested using artus CMV qPCR. Aptima CMV lower limit of detection (LLOD) was 169IU/mL for ACD plasma, 100IU/mL for BAL, and 50IU/mL for tissue. Positive percent agreement (PPA) was 100.0% (50/50; 95% CI: 92.9% - 100.0%) and negative percent agreement (NPA) was 94.0% (47/50; 95% CI: 83.5% - 98.8%) for ACD plasma. Bland-Altman analysis revealed a bias of 0.20log10 IU/mL (Aptima - artus) with 95% limits of agreement of -0.53 to 0.93. For BAL fluids, PPA was 70.0% (14/20; 95% CI: 45.7% - 88.1%) and NPA was 82.4% (43/51; 95% CI: 69.1% - 91.6%). For tissues, PPA was 90.0% (45/50; 95% CI: 78.2% - 96.7%) and NPA was 94.0% (47/50; 95% CI: 83.5% - 98.8%). The Aptima CMV Quant Assay demonstrates high analytical sensitivity and good overall agreement using clinical plasma and tissue specimens.

BIChromET: A Chromogenic Culture Medium for Detection of Piperacillin/Tazobactam and Cefepime Resistance in Pseudomonas aeruginosa.

The BIChromET selective medium for detecting piperacillin-tazobactam (TZP) and cefepime (FEP) resistant Pseudomonas aeruginosa was developed. The performance of this medium was first evaluated using a collection of 100 P. aeruginosa clinical strains (70 TZP-susceptible, 30 TZP-resistant, 58 FEP-susceptible, and 42 FEP-resistant). Then, we performed clinical validation by testing 173 respiratory clinical samples. The BIChromET medium showed excellent sensitivity (TZP (avg. 96.7%); FEP (avg. 92.7%)) and specificity (TZP (avg. 98.9%); FEP (avg. 98%)) in distinguishing the detection limit ranging from 104 to 108 CFU/mL. Then, testing the bronchoalveolar lavage (BAL) and tracheobronchial aspirate (TBA) clinical specimens (N = 173) revealed the excellent performance of the medium with P. aeruginosa, showing 100% and 92.6% of categorical agreements with the results obtained via the broth microdilution methods (BMD) for TZP and FEP, respectively. This medium allows for easy and accurate detection of TZP/FEP-resistant isolates regardless of their resistance mechanisms.

Biofire pneumonia panel in lung donors: faster detection but limited pathogens.

Culture of bronchoalveolar lavage (BAL) specimens takes time to report. We tested whether a molecular diagnostic test could accelerate donor lung assessment and treatment. We compared BioFireFilm ArrayPneumonia Panel (BFPP) with standard of care (SOC) tests on lung allograft samples at three time points: (1) donor BAL at organ recovery, (2) donor bronchial tissue and airway swab at implantation, and (3) first recipient BAL following lung implantation. Primary outcomes were the difference in time to result (Wilcoxon signed-ranked tests) and the agreement in results between BFPP and SOC assays (Gwet's agreement coefficient). We enrolled 50 subjects. In donor lung BAL specimens, BFPP detected 52 infections (14 out of 26 pathogens in the panel). Viral and bacterial BFPP results were reported 2.4 h (interquartile range, IQR 2.0-6.4) following BAL versus 4.6 h (IQR 1.9-6.0, p = 0.625) for OPO BAL viral SOC results and 66 h (IQR 47-87, p < .0001) for OPO BAL bacterial SOC results. Although there was high overall agreement of results between BAL-BFPP versus OPO BAL-SOC tests (Gwet's AC p < .001 for all), the level of agreement differed among 26 pathogens designed in BFPP and differed by types of specimens. BFPP could not detect many infections identified by SOC assays. BFPP decreased time to detection of lung pathogens among donated lungs, but it cannot replace SOC tests due to the limited number of pathogens in the panel.

Quick versus Quantitative: Evaluation of Two Commercial Real-Time PCR Assays for the Detection of Pneumocystis jirovecii from Bronchoalveolar Lavage Fluids.

Two commercial real-time PCR assays for the detection of Pneumocystis jirovecii were compared, the quantitative RealStar P. jirovecii assay and the qualitative DiaSorin P. jirovecii assay, the latter of which can be used without nucleic acid extraction. Archived bronchoalveolar lavage (BAL) specimens (n = 66), previously tested by molecular methods, were tested by both assays, and the results were compared to the respective original result. The RealStar P. jirovecii assay demonstrated good positive percent agreement (PPA) (90% [95% confidence interval (CI), 72 to 97%]; 27/30) and negative percent agreement (NPA) (100% [95% CI, 88 to 100%]; 36/36) with the reference method. The DiaSorin P. jirovecii assay concordantly detected P. jirovecii in 19 of 24 positive BAL samples (PPA = 73% [95% CI, 52 to 88%]). All negative BAL samples gave concordant results (NPA = 100% [95% CI, 87 to 100%]; 34/34). Discordant results occurred mostly in samples with low fungal loads. In conclusion, the RealStar assay demonstrated good concordance with reference results, and the DiaSorin P. jirovecii assay performed well for negative BAL and positive BAL samples with P. jirovecii concentrations of greater than 260 copies/mL. IMPORTANCE Pneumonia, caused by the opportunistic fungus Pneumocystis jirovecii, poses a significant risk for immunocompromised individuals. Laboratory testing for P. jirovecii is progressively shifting toward the use of molecular tests such as real-time PCR; however, this is often performed at reference laboratories. Many frontline laboratories are looking into improving their service and reducing turnaround times for obtaining P. jirovecii results by bringing molecular P. jirovecii testing in-house. We evaluated and compared two commercial real-time PCR assays with different workflows for the detection of P. jirovecii from bronchoalveolar lavage specimens. The RealStar P. jirovecii assay requires nucleic acid extraction and provides a quantification of fungal load for positive samples. The DiaSorin P. jirovecii assay offers a simple workflow without nucleic extraction from patient samples and qualitative results. Results from this study provide valuable information on performance and workflow considerations for laboratories that wish to implement P. jirovecii molecular testing.

Multifacility outbreak of Candida auris during the COVID-19 pandemic—Maricopa County, Arizona, April 2022–December 2022

Background:Candida auris, an emerging and potentially multidrug-resistant fungus, was first identified in Maricopa County, Arizona, in 2020. On April 21, 2022, an acute-care hospital reported C. auris in a bronchoalveolar lavage (BAL) specimen, followed by a second case reported on April 26 identified via retrospective laboratory review and species identification in yeast isolated from a clinical specimen. The Maricopa County Department of Public Health (MCDPH) investigated, and we describe the largest ongoing C. auris outbreak containment response in Maricopa County. Methods: The MCDPH conducted clinical case and contact investigations in accordance with CDC novel organism containment strategy guidelines. In Maricopa County healthcare facilities (HCFs) with suspected transmission, virtual Infection Control Assessment Responses (ICARs) were administered to identify initial infection prevention and control (IPC) gaps; subsequent regular virtual visits were also provided. HCFs with confirmed transmission completed point-prevalence surveys (PPSs) every 2 weeks until transmission halted as evidenced by 2 sequential negative PPSs. Outreach education to affected HCFs was provided to increase awareness about the public health significance of C. auris and the importance of implementation and sustained adherence to standardized IPC protocols. Results: In total, 97 HCFs received IPC outreach education, of which 22 HCFs (23%) had suspected transmission and received a virtual ICAR. Contact investigation identified 1,990 contacts, of whom 1,028 (52%) were discharged to the community, 863 (43%) were admitted to other HCFs, and 99 (5%) died. Of the 863 transferred contacts, 10 (1.2%) declined colonization screening, 853 (98.8%) were screened, and 46 (5%) tested positive for C. auris. Through sequential PPSs, 101 (5%) of 1,914 screened patients tested positive for C. auris. By December 31, 16 clinical and 147 colonized cases were epidemiologically linked to the outbreak. Their median age was 60 years (IQR, 20), and 3 pediatric cases (median age, 17 years) were identified with no pediatric unit admissions. Also, 7colonized cases (5%) developed noninvasive infection and 3 (2%) developed candidemia. Conclusions: The MCDPH’s established partnerships with HCFs were key to this ongoing C. auris outbreak response spanning 22 facilities over 8 months. Challenges included delays in specimen collection and laboratory processing, operational burden of repeated PPS, and ensuring appropriate precautions for readmitted close contacts at subsequent HCFs. The MCDPH assisted facilities in balancing public health surveillance with facility capacity to execute guidance, including repeated PPS. Consistent adherence to stringent IPC practices, interfacility communication, and proactive C. auris education of healthcare workers are paramount to halting transmission.Disclosures: None

Evaluation of BioFire® FilmArray® Pneumonia Panel in Bronchoalveolar Lavage Samples From Immunocompromised Patients With Suspected Pneumonia.

Objectives Immunocompromised patients, specifically those with solid organ transplants or cancer on chemotherapy, are at particularly high risk of severe pneumonia and opportunistic infections. In select patients, bronchoalveolar lavage (BAL) is performed to provide high-quality samples for analysis. We compare BioFire® FilmArray® Pneumonia Panel (BioFire Diagnostics,Salt Lake City, Utah, United States), a multiplex polymerase chain reaction (PCR) assay, with standard of care diagnostics in BAL samples from immunocompromised patients to identify opportunities for this test to affect clinical decision making. Methods Patients hospitalized with pneumonia based on clinical and radiographic findings who underwent evaluation with bronchoscopybetween May 2019 to January 2020 were reviewed. Among those patients undergoing bronchoscopy, those who were immunocompromised were selected for inclusion in the study. BAL specimens submitted to the microbiology laboratory were chosen based onas part of the internal validation of the panel in comparison with sputum culture at our hospitals.We compared the outcomes of the multiplex PCR assay with traditional culture methods and evaluated the role of PCR assay in de-escalating antimicrobial therapy. Results Twenty-four patients were identified for testing with the multiplex PCR assay. Of the 24 patients, 16 were immunocompromised, all with solid or hematological malignancy or a history of organ transplant. Seventeen individual BAL samples from the 16 patients were reviewed. BAL culture results and the multiplex PCR assay were in agreement in 13 samples (76.5%). In four cases, the multiplex PCR assay identified a possible causative pathogen not detected by standard workup. The median time to de-escalation of antimicrobials was three days (interquartile range (IQR) 2-4) from the day of collection of the BAL samples. Conclusions Studies have established the additive role of multiplex PCR testing in addition to traditional diagnostic tools like sputum culture in diagnosing the etiology of pneumonia. Limited data exist specifically looking at immunocompromised patients, in whom a timely and accurate diagnosis is particularly important. There is a potential benefit for performing multiplex PCR assays as an additive diagnostic tool in BAL samples for these patients.

Ureaplasma diversum–Ureaplasma sp. alone or concomitantly with Mycoplasma hyopneumoniae in pig lungs with and without pneumonia: A descriptive exploratory study in abattoirs

Occurrence of Ureaplasma diversum (U. diversum) has been associated with reproductive failures in cattle and detected in pigs with and without pneumonia. However, its role in the porcine respiratory disease complex (PRDC) is unclear. A cross-sectional study was conducted in abattoirs, inspecting 280 pig lungs from eight herds. All the lungs were inspected, processed and classified according to the histopathological analysis. Moreover, bronchoalveolar lavage (BAL) specimens were collected and processed by PCR for detection of U. diversum and Mycoplasma hyopneumoniae (M. hyopneumoniae). Ureaplasma sp.–U. diversum and M. hyopneumoniae were detected in 17.1% and 29.3% of the analyzed BAL specimens, respectively. The concomitant presence of both microorganisms was detected in 12.5% of the inspected lungs. Both agents were found in lungs with and without pneumonia. M. hyopneumoniae was detected in 31.8% of pig lungs with enzootic pneumonia-like lesions, while Ureaplasma sp.–U. diversum was detected in 27.5% of lungs with these lesions. This descriptive exploratory study provides information for future experimental and field-based studies to better define the pathogenic role of this organism within the PRDC.

Gene Expression Study of Host and Mycobacterium tuberculosis Interactions in the Manifestation of Acute Tuberculosis.

Mycobacterium tuberculosis (M.tb) could induce type IV hypersensitivity. The chemotaxis of the leukocytes toward the site of infection and producing matrix metalloproteinases (MMPs) are key factors in the immune pathogenesis of tuberculosis (TB). Mononuclear cells were isolated from bronchoalveolar lavage (BAL) specimens, and the target from genomic DNA was used for qPCR TB diagnosis and cDNA for specific RT-qPCR gene expression. The subjects were then classified into TB+ and TB- groups, and the expression levels of CFP-10, ESAT-6, CCR1, CCR12 and MMP3,9 were evaluated. The mean level of CCR1 expression in TB+ and TB- patients' BAL was 1.71 ± 0.78 and 0.5 ± 0.22, respectively, which was statistically different (p = 0.01). The CCR2 level, in TB+ (2.07 ± 1.4), was higher than in TB- patients (1.42 ± 0.89, p = 0.01). The MMP9 expression in TB+ was 2.56 ± 0.68, also higher than in TB- patients (1.13 ± 0.35), while MMP3 was lower in TB+ (0.22 ± 0.09) than in TB- (0.64 ± 0.230, p = 0.05). The CCR2/CCR1 and MMP3/MMP9 balance in TB+ were reduced, compared to the TB-. The CFP-10 and ESAT-6 were highly expressed in TB+ patients. The CFP-10 expression had a strong negative correlation with albumin (r = - 0.93, p = 0.001), and a negative correlation with neutrophil (r = - 0.444, p = 0.1 with 90% CI). The MMP-9 expression showed a positive correlation with WBC count (r = 0.61, p = 0.02), in TB+, and had a negative correlation with BMI (r = 0.59, p = 0.02) in TB-. The M.tb CFP-10 might be implicated in lowering CCR2 and MMP3 expression in favour of M.tb dissemination. Moreover, the balance of CCR2/CCR1 and MMP3/MMP9 can be used as prognostic factors in the severity of TB.