Detection of Rhizopus-specific antigen in human and murine serum and bronchoalveolar lavage

2020 ◽  
Vol 58 (7) ◽  
pp. 958-964
Author(s):  
Wataru Shibata ◽  
Mamiko Niki ◽  
Kanako Sato ◽  
Hiroki Fujimoto ◽  
Koichi Yamada ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Diagnostic Method ◽  
Signal Sequence ◽  
Specific Antigen ◽  
Invasive Pulmonary Aspergillosis ◽  
Clinical Decision ◽  
Lavage Fluid ◽  
Negative Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa System

Abstract Mucormycosis is a deep-seated fungal infection that mainly develops in patients with severe immunodeficiencies such as those with malignant hematological diseases. Despite poor prognosis, there is no reliable and minimally invasive diagnostic method—such as serodiagnosis—for making a clinical decision regarding the condition. As early diagnosis and early treatment improve the prognosis of mucormycosis, the development of a sensitive early diagnostic method is important. We had previously identified a Rhizopus-specific antigen (RSA) by signal sequence trapping and retrovirus-mediated expression (SST-REX), and evaluated its utility as a diagnostic antigen by constructing a sandwich enzyme-linked immunosorbent assay (ELISA) system to detect serum RSA levels in inoculated mice. In this study, we used the RSA-specific rabbit monoclonal antibodies generated by novel hybridoma technology to improve the sensitivity of the ELISA system. We observed an increase in serum and bronchoalveolar lavage fluid (BALF) levels of RSA in mouse model 1 day after inoculation, suggesting that this newly developed monoclonal antibody-based ELISA system may be useful for the diagnosis of mucormycosis in the early stages of infection. In addition, we measured RSA levels in human serum and BALF, and found that serum RSA level was higher in mucormycosis patients (15.1 ng/ml) than that in invasive pulmonary aspergillosis patients (0.53 ng/ml) and the negative control (0.49 ng/ml). Our results suggest that RSA may be a powerful tool for the diagnosis of pulmonary mucormycosis, and its differentiation from other deep-seated mycoses such as aspergillosis.

scholarly journals Diagnostic Value of Galactomannan Antigen Test in Serum and Bronchoalveolar Lavage Fluid Samples from Patients with Nonneutropenic Invasive Pulmonary Aspergillosis

2017 ◽  
Vol 55 (7) ◽  
pp. 2153-2161 ◽  
Author(s):  
Wei Zhou ◽  
Hongxing Li ◽  
Yan Zhang ◽  
Mei Huang ◽  
Qian He ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Bronchoalveolar Lavage Fluid ◽  
Invasive Pulmonary Aspergillosis ◽  
Lavage Fluid ◽  
Diagnostic Value ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Pulmonary Aspergillosis ◽  
Cutoff Value ◽  
Gm Detection

ABSTRACT The objective of this study was to compare the diagnostic value of galactomannan (GM) detection in bronchoalveolar lavage fluid (BALF) and serum samples from nonneutropenic patients with invasive pulmonary aspergillosis (IPA) and determine the optimal BALF GM cutoff value for pulmonary aspergillosis. GM detection in BALF and serum samples was performed by enzyme-linked immunosorbent assay (ELISA) in 128 patients with clinically suspected nonneutropenic pulmonary aspergillosis between June 2014 and June 2016. On the basis of the clinical and pathological diagnoses, 8 patients were excluded because their diagnosis was uncertain. The remaining 120 patients were diagnosed with either IPA ( n = 37), community-acquired pneumonia (CAP; n = 59), noninfectious diseases ( n = 19), or tuberculosis ( n = 5). At a cutoff optical density index (ODI) value of ≥0.5, the sensitivity of BALF GM detection was much higher than that of serum GM detection (75.68% versus 37.84%; P = 0.001), but there was no significant difference between their specificities (80.72% versus 87.14%; P = 0.286). At a cutoff value of ≥1.0, the sensitivity of BALF GM detection was still much higher than that of serum GM detection (64.86% versus 24.32%; P < 0.001), and their specificities were similar (90.36% versus 95.71%; P = 0.202). Receiver operating characteristic (ROC) curve analysis showed that when the BALF GM detection cutoff value was 0.7, its diagnostic value for pulmonary aspergillosis was optimized, and the sensitivity and specificity reached 72.97% and 89.16%, respectively. BALF GM detection was valuable for the diagnosis of IPA in nonneutropenic patients, and its diagnostic value was superior to that of serum GM detection. The optimal BALF GM cutoff value was 0.7.

scholarly journals The role of Pentraxin3 in plasma and bronchoalveolar lavage fluid in COPD patients with invasive pulmonary aspergillosis

2021 ◽  
Author(s):  
Qian He ◽  
Ming Zhang ◽  
Chunlai Feng
Keyword(s):  
Bronchoalveolar Lavage ◽  
Bronchoalveolar Lavage Fluid ◽  
Invasive Pulmonary Aspergillosis ◽  
Lavage Fluid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasma Samples ◽  
Pulmonary Aspergillosis ◽  
Copd Patients ◽  
Ptx3 Level ◽  
Significant Difference

Abstract BACKGROUND The use of galactomannan testing in plasma and bronchoalveolar lavage fluid (BALF) has improved diagnosis of invasive pulmonary aspergillosis(IPA) in COPD patients; However, the high false positive rate leads to overdiagnosis. This study aimed to investigate the diagnostic value of PTX3 in COPD patients with invasive pulmonary aspergillosis. METHODS A total of 165 patients initially suspected of COPD with invasive pulmonary aspergillosis were included in the study. Among these, 35 cases were proven or probable to be invasive pulmonary aspergillosis (35 plasma samples and 28 BALF samples). The remaining 130 cases were non-aspergillosis controls(130 plasma samples and 83 BALF samples). PTX3 levels and GM were measured by enzyme-linked immunosorbent assay. Results Median plasma and BLAF PTX3 level was significantly higher in COPD patients with invasive pulmonary aspergillosis compared with non-aspergillosis patients (3.74 [2.57–5.61]ng/ml vs 1.29[0.62–2.88] ng/ml,P < 0.001; 3.88[2.28–8.29]ng/ml vs 1.58[0.85–2.13]ng/ml, P < 0.001). When the plasma GM/PTX3 and BALF GM/PTX3 assays were used for patients included in the study, the sensitivity/specificity value were 60%/77.1%/78.6%/89.3%, 73.8%/69.2%/80.7%/77.1%, respectively. Thus, The sensitivity of PTX3 in plasma and BLAF was higher than that of GM. However, There was no significant difference in the specificity of PTX3 and GM between the IPA group and non-aspergillosis group. When PTX3 and GM were both positive in plasma or BLAF, the specificity for the diagnosis of pulmonary aspergillosis can reach more than 90%. Conclusions BALF and plasma PTX3 measurements were significantly higher among patients with IPA. The sensitivity of PTX3 was superior to GM in the diagnosis of IPA in COPD patients. The combination of GM and PTX3 is beneficial to the diagnosis of IPA in COPD patients.

scholarly journals Detection of Anionic Antimicrobial Peptides in Ovine Bronchoalveolar Lavage Fluid and Respiratory Epithelium

1998 ◽  
Vol 66 (12) ◽  
pp. 5948-5954 ◽  
Author(s):  
Kim A. Brogden ◽  
Mark Ackermann ◽  
Kenneth M. Huttner
Keyword(s):  
Bronchoalveolar Lavage ◽  
Genomic Library ◽  
Lavage Fluid ◽  
Maldi Mass Spectrometry ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microbial Infection ◽  
Hybridization Probe ◽  
Pulmonary Tissue ◽  
Western Blots ◽  
Degenerate Oligonucleotide

ABSTRACT Three small antimicrobial anionic peptides (AP) were originally isolated from an ovine pulmonary surfactant. However, their presence in bronchoalveolar lavage (BAL) fluid and tissues of the respiratory tract is unknown. In this study, we made affinity-purified rabbit polyclonal and mouse monoclonal antibodies to synthetic H-DDDDDDD-OH. Antibody specificity was assessed by a competitive enzyme-linked immunosorbent assay (ELISA), and the exact epitope binding sites were determined with analog peptides synthesized on derivatized cellulose. These antibodies were used to detect AP in BAL fluid by ELISA and in respiratory tissues by Western blot analysis and immunocytochemistry. BAL fluid from 25 sheep contained 0.83 ± 0.33 mM AP (mean ± standard deviation; range, 0.10 to 1.59 mM) and was antimicrobial. The presence of AP in BAL fluid was confirmed by reverse-phase high-pressure liquid chromatography fractionation followed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry on those fractions which were positive by competitive ELISA and demonstrated antimicrobial activity. In Western blots, polyclonal antibody PAB96-1 and monoclonal antibody 1G9-1C2 (5.0 μg/ml) detected four bands in solubilized turbinate and tracheal epithelial cells (53.7, 31.2, 28.0, and 25.7 kDa) and five bands in lung homogenates (53.5, 37.1, 31.2, 28.0, and 25.7 kDa). Only a single band was seen in solubilized liver and small-intestine homogenates, and no bands were seen in blots containing BAL fluid, albumin, or kidney or spleen homogenates. In pulmonary-tissue sections, both antibodies PAB96-1 and 1G9-1C2 identified accumulated protein in the apical cytoplasm of the bronchial and bronchiolar epithelia, in the cytoplasm of pulmonary endothelial cells, and in an occasional alveolar macrophage. As a first step in identifying a candidate AP precursor gene(s), degenerate oligonucleotides representing all possible coding combinations for H-GADDDDD-OH and H-DDDDDDD-OH were synthesized and used to probe Southern blots of sheep genomic DNA. Following low-stringency washes and a 2-day exposure, strongly hybridizing bands could be identified. One degenerate oligonucleotide, SH87, was used as a hybridization probe to screen a sheep phage genomic library. Two independent phage contained the H-GADDDDD-OH coding sequence as part of a larger predicted protein. AP may originate as part of an intracellular precursor protein, with multistep processing leading to the release of the heptapeptide into mucosal secretions. There it may interact with other innate pulmonary defenses to prevent microbial infection.

scholarly journals Decreased Production of Local Immunoglobulin A toPneumocystis carinii in Bronchoalveolar Lavage Fluid from Human Immunodeficiency Virus-Positive Patients

2000 ◽  
Vol 68 (3) ◽  
pp. 1054-1060 ◽  
Author(s):  
A. Jalil ◽  
P. Moja ◽  
C. Lambert ◽  
M. Perol ◽  
L. Cotte ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Bronchoalveolar Lavage ◽  
Bronchoalveolar Lavage Fluid ◽  
Pneumocystis Carinii ◽  
Immunoglobulin A ◽  
Lavage Fluid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Local Production ◽  
Immunodeficiency Virus ◽  
Negative Controls

ABSTRACT An enzyme-linked immunosorbent assay and a Western blot analysis were developed to study the antibody response to Pneumocystis carinii in serum and bronchoalveolar lavage fluid from 27 human immunodeficiency virus 27 (HIV)-infected patients with P. carinii pneumonia (Pcp), 32 patients without Pcp, and 51 HIV-negative controls. Urea was used for the correct dilution of epithelial lining fluid, and albumin was used to evaluate transudation from plasma for the assessment of local production of antibodies toP. carinii. By contrast with those of immunoglobulin G (IgG), IgA responses to P. carinii were increased in serum from HIV-positive patients compared to negative controls. Local production of antibodies to P. carinii, especially IgA, was decreased in patients with Pcp. In a study of 10 patients of each group, IgG and IgA responses to gp116 from P. carinii were lower in patients with Pcp than in other groups. These results suggest that, in addition to alveolar macrophages, local antibodies may play a role in host defense against P. carinii.

scholarly journals Early protein expression profile in bronchoalveolar lavage fluid and clinical outcomes in primary graft dysfunction after lung transplantation

2020 ◽  
Vol 58 (2) ◽  
pp. 379-388
Author(s):  
Anna E Frick ◽  
Stijn E Verleden ◽  
Sofie Ordies ◽  
Annelore Sacreas ◽  
Robin Vos ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Lung Transplantation ◽  
Lung Transplant ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transplant Recipients ◽  
Primary Graft Dysfunction ◽  
Graft Dysfunction ◽  
Lung Transplant Recipients

Abstract OBJECTIVES Primary graft dysfunction (PGD) remains a major post-transplant complication and is associated with increased morbidity and mortality. Mechanisms evoking PGD are not completely clear, but inflammation plays a central role. We investigated the association between PGD and inflammatory proteins present in immediate postoperative bronchoalveolar lavage. METHODS All double-lung recipients transplanted at our institution from 2002 to 2018 were included in our study. We retrospectively selected 80 consecutive lung transplant recipients with different PGD grades (n = 20 for each PGD grades 0–1 to 2–3). In bronchoalveolar lavage performed within the first 24 h after donor aortic cross-clamping following lung transplantation, concentrations of 30 cytokines, chemokines and growth factors were assessed by enzyme-linked immunosorbent assay (ELISA) and correlated with donor and recipient demographics and outcomes. For analysis, 2 groups were defined: ‘mild’ PGD (grade 0–1) and ‘severe’ PGD (grades 2–3). RESULTS Significant differences between mild and severe PGD were found in 8 biomarkers [interleukin (IL)-6, IL-10, IL-13, eotaxin, granulocyte colony-stimulating factor, interferon γ, macrophage inflammatory protein 1α, surfactant protein D (SP-D); P &lt; 0.05]. Increased IL-10 and IL-13, but none of the other proteins, were associated with short-term outcome (longer time to extubation; P = 0.005 and P &lt; 0.0001; increased intensive care unit stay; P = 0.012 and P &lt; 0.0001; and hospital stay; P = 0.041 and P = 0.002). There were no significant differences in donor and recipient characteristics between the groups. CONCLUSIONS Expression profiles of key inflammatory mediators in bronchoalveolar lavage fluid differed significantly between lung transplant recipients with severe versus mild PGD and correlated with clinical outcome variables. Further research should focus on the early mechanisms leading to PGD.

scholarly journals Galactomannan in Bronchoalveolar Lavage Fluid for Diagnosis of Invasive Pulmonary Aspergillosis with Nonneutropenic Patients

2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Qidong Zhuang ◽  
Hongying Ma ◽  
Yun Zhang ◽  
Lei Chen ◽  
Li Wang ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Bronchoalveolar Lavage Fluid ◽  
Invasive Pulmonary Aspergillosis ◽  
Final Analysis ◽  
Lavage Fluid ◽  
Control Group ◽  
Control Analysis ◽  
Pulmonary Aspergillosis ◽  
Significant Difference ◽  
Gm Detection

Background. We evaluated the utility of galactomannan (GM) in bronchoalveolar lavage fluid (BALF) for the diagnosis of invasive pulmonary aspergillosis (IPA) in nonneutropenic patients. Methods. A total of 183 patients were included in the final analysis. Bronchoscopies and the detection of GM in BALF were all performed on them. Results. Ten cases of IPA were diagnosed. ROC data demonstrated that, for diagnosing IPA, an optimal cutoff value for GM in BALF of 0.76 yielded a sensitivity of 100.0% and a specificity of 76.2%. Symptoms and radiological findings had no significant difference between proven or probable IPA group and non-IPA group. In our case-control analysis, although nine patients with false-positive results received treatment with Piperacillin/tazobactam, there was no significant difference between case and control group. Conclusions. BALF GM detection is a valuable adjunctive diagnostic tool. Our retrospective study suggests that the optimal value of GM detection in BALF is 0.76 in nonneutropenic patients.

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