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Traditional laboratory detection methods and metagenomic next-generation sequencing in pulmonary fungal infection diagnosis

In recent years, due to the extensive usage of immunosuppressant and the rise of patients with cancers and organ transplantation, the incidence rate of invasive fungal infection, especially invasive pulmonary fungal infection, has increased. Besides the clinical manifestations, medical history and imaging, the diagnosis of pulmonary mycosis mainly depends on pathogen detection methods in clinical microbiology laboratory. However, due to the difficulty in fungi culturing and the low sensitivity of smear microscopy, better molecular biology methods are needed. To date, the emergence of metagenomic next-generation sequencing (mNGS) has improved the identification rate of pulmonary fungal infections. mNGS is significantly superior to traditional detection methods in rapid, accurate, and comprehensive determination of fungi from various clinical specimens, especially atypical fungi. However, some problems in mNGS method have to be addressed including sample collection, report interpretation, and its combination with traditional microbiology methods. With the in-depth discussion and solution of the above problems, mNGS will be indispensable to the etiological diagnosis of pulmonary invasive fungal infection.

The Interpretation of Guidelines for the Management of Candidiasis from Infectious Diseases Society of America 2009

In vitro Antifungal Spectrum and Activity of 1% Naftifine-0.25% Ketoconazole Cream Compared with 2% Ketoconazole Cream and 1% Terbinafine Cream

Objective　To research the anti-fungal spectrum and activity of the cream containing 1% naftifine-0.25% ketoconazole compared with other two creams that contain of 2% ketoconazole and of 1% terbinafine, respectively. Methods　The agar diffusion method was used to judge drug sensitivity. Twenty-nine isolates of pathogenic fungi belonging to 11 species from clinic and three species of Malassezia standard stains were enrolled into the experiment. Organism suspension of each species was spread on the surface of the plate of the optimal media containing 2% agar. Then wells were made in the plate and three types of cream were put in each well respectively. After seven-day incubation, the diameter of the inhibition zone around the well full of each cream was observed and recorded. Results　The inhibition zone around the well full of 1% naftifine-0.25% ketoconazole cream for all experiment isolates (Dermatophytes, Candida spp., Sporothrix schenkii, Fonsecaea pedrosoi, Fusarium graminearum, Malassezia furfur, M. globosa and M. sympodialis) was observed, with the mean diameter of 45.46mm. Similarly, the mean diameter of inhibition zone of 2% ketoconazole cream for all experiment isolates was 23.92mm. About 1% terbinafine cream, the mean diameter was 29.81mm but there was no inhibition zone observed around Candida krusei and Candida albicans mycelial-form. There were significant significances for mean diameters of the inhibition zone when comparing 1% naftifine-0.25% ketoconazole cream with 2% ketoconazole cream (P=0.000) and with 1% terbinafine cream (P=0.000). Conclusion　The anti-fungal spectrum of 1% naftifine-0.25% ketoconazole cream is wider than that of 1% terbinafine cream. The antifungal activity of 1% naftifine-0.25% ketoconazole cream is ber than that of 2% ketoconazole cream and 1% terbinafine cream.

The value of 1,3-beta-D-glucan assay for diagnosis of invasive fungal disease with automimmune disease

ObjectiveTo evaluate the diagnostic value of monitoring 1,3-beta-D-glucan (G test) in patients with autoimmune disease complicated with invasive fungal disease (IFD). MethodsA retrospective study was performed in hospitalized patients in the First Affiliated Hospital of Zhengzhou Universisty who were diagnosed as autoimmune disease with lung infection during the immunosuppressive therapy between January 2014 and January 2016. A total of 372 patients were enrolled in this study. All subjects were classified according to the 2006 diagnostic criteria and treatment of invasive pulmonaary fungal infection, with serum 1,3-β-D-glucan results not included in the diagnosis. There were 18 cases with proven IFD, 35 cases with probable IFD, and 70 ceses with possible IFD. Fifty-three patients with proven IFD or probable IFD were as a case group, and another 249 patients with no evidence for IFD were as a control group. The value of the G test for diagnosis of automimmune disease with IFD was analyzed by ROC curve. ResultsThe serum 1,3-β-D-glucan level was significantly higher in the case group when compared with the control group [median (interquartile range): 135.0 (63.1 to 319.0) pg/ml vs. 75.9 (41.2 to 88.1) pg/ml, P<0.05]. When the cut-off value of serum 1,3-β-D-glucan level was set at 93.8 pg/ml, the sensitivity, specificity, positive predictive value, and negative predictive value for diagnosis of autoimmune disease with IFD were 0.65 (95% CI 0.56 to 0.73), 0.87 (95% CI 0.83 to 0.92), 0.70 (95% CI 0.64 to 0.81), and 0.83 (95% CI 0.79 to 0.88), respectively. ConclusionThe 1,3-beta-D-glucan test is a valuable method for diagnosis of IFD in patients with autoimmune disease.

Risk Factors of Invasive Fungal Infection in Respiratory Ward: A Retrospective Case Control Study

Objective To explore the risk factors of invasive fungal infection ( IFI) in respiratory ward. Methods A multi-center, retrospective, case-control study was carried out. Patients from five general hospitals in Chongqing city, diagnosed as fungal infection, or whose respiratory specimens were fungal positive, were retrospectively screened for IFI. Patients with respiratory infection and colonization of nonfungal cases in the same period of hospitalization were enrolled as control. Results Thirty-four patients diagnosed with IFI and 50 patients diagnosed with bacterial infection were analyzed for the risk factors of IFI. The demographic characteristics of patients including age and gender were not different( P gt; 0. 05) , but hospitalization days, carbapenem antibiotic use, chemotherapy, deep venous catheterization, total parenteralnutrition( TPN) , neutropenia, and renal disfunction were different significantly between the IFI group and the control group. Multiple logistic regression analysis showed that carbapenem antibiotic use ( OR = 6. 753) ,central venous catheterization ( OR = 5. 021) and TPN ( OR = 3. 199) were main risk factors of invasive fungal infection. Conclusion The carbapenem antibiotic use, central venous catheterization and TPN are risk factors for IFI in respiratory ward.

Clinical Research on Correlation Factors of Concurrent Fungal Infections in Old Patients with Pulmonary Diseases: 49 Case Reports

摘要:目的: 探討我院呼吸內科病房老年肺部疾病患者并發真菌感染發病的相關因素,分析其易患因素、臨床特征和治療。 方法 : 采用回顧性調查方法對2002年1月至2008年6月收住內科的經微生物檢查證實49例繼發真菌感染的患者進行分析,并與同期無真菌感染的肺部疾病患者(對照組)比較。 結果 : 在呼吸內科病房中,老年患者院內肺部真菌感染發生率為378%,主要感染部位為泌尿系(218%),呼吸道(269%),消化道(409%)。慢性阻塞性肺疾病(498%)是繼發院內肺部真菌感染最常見的基礎疾病,其感染因素為長期使用廣譜抗生素(962%)和糖皮質激素(332%)、營養狀況不良(583%)出現低蛋白血癥及合并糖尿病、白細胞減少和侵襲性診療操作等。肺部真菌感染的臨床表現無特異性,確診需結合痰培養,組織病理學和臨床表現來確定,感染菌種以白色念珠菌為主,占626%。氟康唑治療有效率914%。研究組與同期無真菌感染的肺部疾病患者(對照組)比較:病死率分別為612%和082%,兩組治療無效的病例(惡化和死亡病例)比較差異有顯著性。 結論 : 院內真菌是呼吸系統疾病繼發感染的重要病原體,而白色假絲酵母菌是院內肺部真菌感染的主要致病菌,宿主免疫狀態、感染播散和疾病嚴重程度是影響預后的因素。該研究認為老年肺部疾病患者并發真菌感染的相關因素和影響預后的因素對其預防、診斷、治療、改進預后和生存質量有重要的臨床意義。除有效的抗真菌治療外,積極的綜合治療有助于提高真菌感染的治愈率。Abstract: Objective: To study the susceptible factors,clinical features and treatments of nosocomial pulmonary fungal infection in the ward of respiratory department.〖WTHZ〗Methods : The chart files of 49 patients with nosocomial pulmonary fungal infection admitted from January 2002 to June 2008 in the ward of Respiratory Department were reviewed. Results : The incidence rate of nosocomial pulmonary fungal infection was 378%.COPD(498%)was the main predisposing disease,and candidiasis(626%) was the most common pathogen. The main susceptible factors associated with nosocomial pulmonary fungal infection are longterm use of broadspectrum antibiotics(962%),hypoalbuminemia(583%),longterm use of adrenocortical steroid(332%),and diabetes mellitus.There is no specific clinical feature.Fluconazole(914%)is more efficient in the treatment.〖WTHZ〗Conclusion : Nosocomial pulmonary fungis are important pathogenin the secondary infection in respiratory disease.The most common pathogen is candida albicans.Combined therapy as well as treating fungus infection are important measures to increase the cure rate of nosocomial pulmonary fungal infection.

The clinical features and prognostic analysis of visual function damage caused by fungal sphenoid sinusitis

ObjectiveTo observe the clinical characteristics of patients with visual impairment caused by fungal sphenoid sinusitis and analyze the influencing factors related to visual prognosis. Methods A retrospective clinical study. From January 2006 to December 2020, 44 patients (55 eyes) with visual impairment caused by fungal sphenoid sinusitis confirmed by imaging and pathological examination in the Department of Ophthalmology of Beijing Tongren Hospital were included in the study. Patients was first diagnosed in the Department of Ophthalmology due to monocular or binocular vision loss, or binocular diplopia, limited eye movement and ptosis. All patients underwent visual acuity examination and fundus color photography. CT examination of paranasal sinus or orbit was performed in 37 cases; magnetic resonance imaging (MRI) of paranasal sinus, brain or orbit was performed in 34 cases. All patients underwent endoscopic sinus opening combined with intrasinus lesion clearance; 14 cases were treated with antifungal drugs after operation. The average follow-up time was 59.61±37.70 months. Comparison of clinical characteristics between invasive and non-invasive fungal sphenoid sinusitis were by χ2 test or Fisher exact test. The influencing factors with P＜0.2 in univariate analysis were selected for multivariate regression analysis. ResultsAmong the 44 patients, there were 19 males and 25 females; the ratio of male to female was 1:1.3; the average age of visual symptoms was 61.48 ± 12.17 years; 23 cases (52.3%, 23/44) suffered from immune dysfunction, including 21 cases of diabetes mellitus. The visual acuity decreased in 33 cases (44 eyes) (75.0%, 33/44). There were 15 cases of binocular diplopia with eye movement disorder (34.0%, 15/44), including 6 cases with visual impairment. The visual acuity of the affected eye was no light perception-0.8. There were 35 cases with headache (79.5%, 35/44). Nasal symptoms were found in 14 cases (31.8%, 14/44). There were 40 and 4 cases of Aspergillus and Mucor infection in sphenoid sinus, respectively. Among the 37 cases who underwent CT examination of paranasal sinus or orbit, there were soft tissue filling in the sinus cavity, including 19 cases of high-density calcification in the sinus cavity (51.4%, 19/37); bone defect of sinus wall were in 24 cases (64.9%, 24/37). There were 26 cases (70.3%, 26/37) of sinus wall osteosclerosis. MRI of paranasal sinus, brain or orbit was performed in 34 cases. T1WI of sphenoid sinus lesions showed low signal, high signal and equal signal in 14, 10 and 9 cases, respectively; T2WI showed high signal, low signal and equal signal in 13, 16 and 2 cases respectively. After enhancement, the lesions were strengthened in 11 cases, no obvious enhancement in 23 cases, and the surrounding mucosa was thickened and strengthened. The lesions involved the orbital apex and cavernous sinus in 18 and 16 cases, respectively; orbital apex and cavernous sinus were involved in 12 cases. Six months after operation, visual acuity was significantly improved in 27 eyes (65.9%, 27/41); visual acuity did not improve in 14 eyes (34.1%, 14/41). Multivariate regression analysis showed that the change of sinus wall osteosclerosis was associated with higher visual acuity improvement rate (odds ratio= 0.089, 95% confidence interval 0.015-0.529, P=0.008). ConclusionsFungal sphenoid sinusitis related visual impairment is relatively common in elderly female patients with low immune function; monocular vision loss with persistent headache is the most common clinical symptom; imaging findings of sphenoid sinus lesions are an important basis for diagnosis. Sphenoid sinus opening combined with sinus lesion clearance is an effective treatment. After operation, the visual acuity of most patients can be improved. The prognosis of visual acuity was relatively good in patients with hyperplasia and sclerosis of sphenoid sinus wall bone.

Discussion on the construction of fungal disease prevention and control system in China from the epidemic of Candida auris

Candida auris is an emerging multidrug-resistant fungus that has become a significant global public health threat due to its strong resistance to antifungal agents and its ability to spread within healthcare facilities. This paper reviews the global epidemiological trends of Candida auris and the current status of existing prevention and control systems, focusing specifically on pathogen epidemiological characteristics, domestic and international epidemic situations, current prevention and control frameworks, and the construction of prevention networks. In response to the challenges posed by the international spread of Candida auris , China’s fungal disease prevention system should advance towards a more systematic and scientific direction. By integrating resources from medical institutions, disease control agencies, and research institutes, and combining multidisciplinary knowledge and technologies, China should establish a multi-level coordinated prevention and control mechanism to improve its monitoring, prevention, and treatment systems. In the future, China’s fungal disease prevention and control system needs to further strengthen talent cultivation, improve surveillance networks, promote technological innovation, and build a comprehensive, multi-level modern prevention and control system.

Clinical characteristics and pathogen distribution of patients with community-acquired pneumonia and type 2 diabetes mellitus

Objective To investigate the clinical characteristics and pathogen distribution of community-acquired pneumonia (CAP) combined with type 2 diabetes mellitus (T2DM), based on bronchoalveolar lavage fluid (BALF) metagenomic next-generation sequencing (mNGS) test. Methods In this cross-sectional study, CAP patients with BALF mNGS test were screened from April 2023 to April 2024. The patients were divided into a single CAP group (CAP group) and a CAP combine with T2DM group (CAP+T2DM group). The data of demographics, underlying diseases, complications, and laboratory tests including blood routine, inflammatory parameters, liver and renal functions, random blood glucose (RGB), hemoglobin A1C (HbA1c), and BALF mNGS tests were collected and compared between the two groups. Results Ultimately, 86 patients were included, with 45 in the CAP group and 41 in the CAP+T2DM group. Compared with the CAP group, the CAP+T2DM group had higher platelet count [(272.44±128.57)×109/L vs. (215.00±100.06)×109/L], erythrocyte sedimentation rate [(75.63±35.19) vs. (59.69±34.47) mm/h], RGB [10.8 (9.1, 13.5) vs. 6.5 (5.8, 7.8) mmol/L], HbA1c [8.2% (7.3%, 8.5%) vs. 5.7% (5.5%, 6.1%)], and fungi infection rate (65.9% vs. 40.0%), and the differences were statistically significant between the two groups (P<0.05). Conclusion CAP patients with T2DM have increased levels of platelet and erythrocyte sedimentation rate, and are at higher risk for fungi infection, which potentially leads to worse outcome.

A nomogram for predicting secondary pulmonary fungal infection in patients with acute exacerbations of chronic obstructive pulmonary disease

Objective To investigate the risk factors for secondary pulmonary fungal infection in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). And a visual tool using nomogram was developed and validated to assist in the clinical prediction of the probability of pulmonary fungal infection occurrence in AECOPD patients. Methods A retrospective cohort study method was used to collect AECOPD patients hospitalized in the Department of Respiratory, The First Affiliated Hospital of Chengdu Medical College from January 2021 to December 2021 as a training set. And AECOPD patients between January 2020 and December 2020 were collected as a validation set. Independent risk factors were determined through univariate, Lasso regression analyses. and multivariable logistic, A nomogram prediction model was constructed with these independent risk factors, and the nomogram was evaluated by receiver operating characteristic area under the curve (AUC), calibration curve, and decision curve analysis (DCA). Results The use of glucocorticoid, combined use of antibiotics, duration of antibiotic use and hypoalbuminemia were independent risk factors for secondary pulmonary fungal infection in AECOPD patients (all P<0.05). The training set and validation set of the constructed prediction model had an AUC value of 0.915 [95%CI: 0.891 - 0.940] and 0.830 [95%CI: 0.790 - 0.871], respectively. The calibration curve showed that the predicted probability was in good agreement with the actual observed probability of pulmonary fungal infection in AECOPD patients. The corresponding decision curve analysis (DCA) indicated the nomogram had relatively ideal clinical utility. Conclusions The result showed that the use of glucocorticoid, combined use of antibiotics, prolonged antibiotic therapy and hypoalbuminemia was independent risk factors for pulmonary fungal infection in AECOPD patients. The clinical prediction model for secondary pulmonary fungal infection in AECOPD patients constructed in this study has strong predictive power and clinical practicability.