scholarly journals Next-generation sequencing technology for detecting pulmonary fungal infection in bronchoalveolar lavage fluid of a patient with dermatomyositis: a case report and literature review

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Kaiyu Zhang ◽  
Chen Yu ◽  
Yuxiang Li ◽  
Yang Wang
2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Suguru Takeuchi ◽  
Jun-ichi Kawada ◽  
Kazuhiro Horiba ◽  
Yusuke Okuno ◽  
Toshihiko Okumura ◽  
...  

Abstract Next-generation sequencing (NGS) has been applied in the field of infectious diseases. Bronchoalveolar lavage fluid (BALF) is considered a sterile type of specimen that is suitable for detecting pathogens of respiratory infections. The aim of this study was to comprehensively identify causative pathogens using NGS in BALF samples from immunocompetent pediatric patients with respiratory failure. Ten patients hospitalized with respiratory failure were included. BALF samples obtained in the acute phase were used to prepare DNA- and RNA-sequencing libraries. The libraries were sequenced on MiSeq, and the sequence data were analyzed using metagenome analysis tools. A mean of 2,041,216 total reads were sequenced for each library. Significant bacterial or viral sequencing reads were detected in eight of the 10 patients. Furthermore, candidate pathogens were detected in three patients in whom etiologic agents were not identified by conventional methods. The complete genome of enterovirus D68 was identified in two patients, and phylogenetic analysis suggested that both strains belong to subclade B3, which is an epidemic strain that has spread worldwide in recent years. Our results suggest that NGS can be applied for comprehensive molecular diagnostics as well as surveillance of pathogens in BALF from patients with respiratory infection.


2021 ◽  
Author(s):  
Jiangqin Song ◽  
Fuying Hu ◽  
Huashun Li ◽  
Youping Chen ◽  
Weifang Zhu ◽  
...  

Abstract BackgroundMucor infection cannot be ignored in patients with pulmonary shadowing with cavitation .This paper reports a case of mucormycosis during the COVID-19 pandemic in Hubei Province, China. Case PresentationA anesthesiology doctor was initially diagnosed as COVID-19 due to changes in lung imaging. Later Lichtheimia ramose was found by Metagenomic next generation sequencing (mNGS) in the Bronchoalveolar lavage fluid (BALF).After adjusting amphotericin B for anti-infective treatment, the patient's infection lesions were shranked and the symptoms were significantly relieved. ConclusionThe diagnosis of invasive fungal infections is very difficult, mNGS can make an accurate pathogenic diagnosis of invasive fungal diseases for the clinic and provide a basis for clinical treatment.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Meiqin Chen ◽  
Weili Lu ◽  
Shugen Wu ◽  
Shun Wang ◽  
Tao Lu ◽  
...  

Abstract Background Leptospirosis is a common infectious disease in tropical and semitropical regions, and it is typically neglected. Leptospirosis-associated acute diffuse alveolar hemorrhage is one of its fatal complications. The use of bronchoalveolar lavage fluid (BALF) metagenomic next-generation sequencing in the diagnosis of Leptospira interrogans infection has rarely been reported. Case presentation We present the case of a 62-year-old female who was transferred to our hospital with dyspnea, and severe hemoptysis and was supported by a tracheal intubation ventilator. Bronchoalveolar lavage fluid (BALF) metagenomic next-generation sequencing (mNGS) reported Leptospira interrogans. A diagnosis of diffuse alveolar hemorrhage caused by leptospirosis was made. After immediately receiving antibiotics and hormone therapy, the patient achieved a complete recovery upon discharge. Conclusion Leptospirosis presenting as severe diffuse alveolar hemorrhage is rare but should be considered in the differential diagnosis. mNGS can help identify pathogens and treat them early, which can improve prognosis.


2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Wanghui Shi ◽  
Shanshan Zhu

Objective. To uncover the application value of metagenomic next-generation sequencing (mNGS) in the detection of pathogen in bronchoalveolar lavage fluid (BALF) and sputum samples. Methods. Totally, 32 patients with pulmonary infection were included. Pathogens in BALF and sputum samples were tested simultaneously by routine microbial culture and mNGS. Main infected pathogens (bacteria, fungi, and viruses) and their distribution in BALF and sputum samples were analyzed. Moreover, the diagnostic performance of mNGS in paired BALF and sputum samples was assessed. Results. The pathogen culture results were positive in 9 patients and negative in 13 patients. No statistical differences were recorded on the sensitivity (78.94% vs. 63.15%, p = 0.283 ) and specificity (62.50% vs. 75.00%, p = 0.375 ) of mNGS diagnosis in bacteria and fungus in two types of samples. As shown in mNGS detection, 10 patients’ two samples were both positive, 13 patients’ two samples were both negative, 7 patients were only positive in BALF samples, and 2 patients’ sputum samples were positive. Main viruses mNGS detected were EB virus, human adenovirus 5, herpes simplex virus type 1, and human cytomegalovirus. Kappa consensus analysis indicated that mNGS showed significant consistency in detecting pathogens in two samples, no matter bacteria ( p < 0.001 ), fungi ( p = 0.026 ), or viruses ( p = 0.008 ). Conclusion. mNGS showed no statistical differences in sensitivity and specificity of pathogen detection in BALF and sputum samples. Under certain conditions, sputum samples might be more suitable for pathogen detection because of invasiveness of BALF samples.


2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S658-S659
Author(s):  
Suguru Takeuchi ◽  
Jun-ichi Kawada ◽  
Kazuhiro Horiba ◽  
Yusuke Okuno ◽  
Toshihiko Okumura ◽  
...  

Abstract Background In the field of infectious diseases, identification of etiologic pathogen is essential for definitive diagnosis and decisions regarding appropriate management. Bronchoalveolar lavage fluid (BALF) is considered a sterile type of specimen that is suitable for detecting pathogens of respiratory infections. Recently, next-generation sequencing (NGS) has been applied in the field of infectious diseases and has enabled us to identify pathogenic microorganisms comprehensively. The aim of this study was to comprehensively identify pathogens using NGS in BALF samples from immunocompetent pediatric patients with respiratory failure. Methods Ten patients hospitalized in the pediatric intensive care unit with respiratory failure were included. BALF samples obtained in the acute phase were used to prepare DNA- and RNA-sequencing libraries. The libraries were sequenced on MiSeq, and the sequence data were analyzed using metagenome analysis tools. Results A mean of 2,041,216 total reads were sequenced for each library. A significant number of four types of bacterial reads was detected in three BALF samples with DNA-sequencing, whereas pathogenic respiratory viruses were detected in seven of 10 patients with RNA-sequencing.Candidate pathogens were detected in three patients in whom etiologic agents were not identified by conventional methods. A summary of the detected pathogens is listed in Table 1. Sequence coverage and depth of each reference bacterial and viral genome are shown in Figures 1 and 2, respectively. The complete genome of enterovirus D68 was identified in two patients without underlying diseases, and phylogenetic analysis suggested that both strains belong to subclade B3, which is an epidemic strain that has spread worldwide in recent years. Conclusion We demonstrated the utility of the NGS-based approach for detection of candidate pathogens in BALF from pediatric patients with severe respiratory failure. Our results suggest that NGS can be applied for comprehensive molecular diagnostics as well as surveillance of pathogens in the field of infectious diseases. Disclosures All authors: No reported disclosures.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Lu Qin ◽  
Fei-Zhou Zhang ◽  
Tong-Yu Yang ◽  
Xiao-Fen Tao ◽  
Lan-Fang Tang

Abstract Background Idiopathic pulmonary hemosiderosis (IPH) encompasses a rare and agnogenic group of diffuse alveolar capillary hemorrhagic diseases. Corticosteroid treatment is the globally preferred therapeutic strategy for IPH; however, it can cause immunodeficiency. Nocardia infection often occurs in immunocompromised patients and primarily involves the pleura and lungs. Herein, we describe a case of pediatric pulmonary Nocardia infection after the corticosteroid treatment of IPH. Case presentation A 7-year-old girl presented with chief complaints of pale complexion persisting for 1 year and a cough for 20 days. Abundant hemosiderin-laden macrophages were detected in the gastric juice, which supported the diagnosis of IPH. Uninterrupted doses of corticosteroids were administered during the last hospitalization. After nearly 2 months of corticosteroids therapy, the patient began to cough and produce a purulent sputum. Next-generation sequencing of the bronchoalveolar lavage fluid revealed Nocardia abscessus (N. abscessus) DNA. Linezolid was administered with good response, and the patient was discharged after 18 days of hospitalization. Her symptoms and pulmonary lesions had recovered, and the IPH appeared to be well-controlled with low dose of corticosteroids in follow-up. Conclusions Nocardia infection should be considered in the differential diagnoses for IPH patients receiving corticosteroid therapy, especially in patients with poor response to conventional empirical antibiotic therapy. Next-generation sequencing of bronchoalveolar lavage fluid may be used to quickly identify the Nocardia. Sulfonamides or linezolid are effective for pediatric pulmonary Nocardia infection.


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