Fungal Diagnosis Research Articles

Cross-sectional investigation of mycological diagnosis challenges in Saudi Arabia.

The global incidence of fungal infection has increased dramatically over the last two decades. Fungal diseases threaten both immunocompetent, and immunocompromised patients. The current fungal diagnostics status in Saudi Arabia needs to be evaluated, especially with the increase of the immunosuppressed population. This cross-sectional study investigated the gaps in mycological diagnosis on a national level. The call interview questionnaire responses were collected to evaluate the demand for fungal assays, diagnostic methods' quality, and mycological expertise of laboratory technologists in both public and private medical intuitions. The data were analyzed using (IBM SPSS ® software version 22.0). A total of 57 hospitals from all Saudi regions participated in the questionnaire; however, only 32% received or processed mycological samples. Most participants were from the Mecca region (25%), Riyadh region (19%), and Eastern region (14%). The top fungal isolates identified were Candida spp., Aspergillus spp., and dermatophyte. Fungal investigation is highly requested by intensive care, dermatology, and obstetrics and gynecology units. Most laboratories rely on fungal culture and microscopic examination, which mostly identify Candida to the genus level, and use 37°C incubators for culture (67%). Antifungal susceptibility testing (AST) and serological and molecular methods are rarely performed and mostly outsourced. Using accurate identification and AST are the primary factors to improve fungal diagnosis in respect to turnaround time and cost. The three major obstacles identified were availability of facility (47%), reagents and kits (32%), and good training (21%). The results indicated that fungal diagnosis demand was relatively higher in high-population regions. This study highlighted the gaps in fungal diagnostics reference laboratories to encourage their improvement in Saudi hospitals.

Comparisons of artificial intelligence algorithms in automatic segmentation for fungal keratitis diagnosis by anterior segment images.

This study combines automatic segmentation and manual fine-tuning with an early fusion method to provide efficient clinical auxiliary diagnostic efficiency for fungal keratitis. First, 423 high-quality anterior segment images of keratitis were collected in the Department of Ophthalmology of the Jiangxi Provincial People's Hospital (China). The images were divided into fungal keratitis and non-fungal keratitis by a senior ophthalmologist, and all images were divided randomly into training and testing sets at a ratio of 8:2. Then, two deep learning models were constructed for diagnosing fungal keratitis. Model 1 included a deep learning model composed of the DenseNet 121, mobienet_v2, and squeezentet1_0 models, the least absolute shrinkage and selection operator (LASSO) model, and the multi-layer perception (MLP) classifier. Model 2 included an automatic segmentation program and the deep learning model already described. Finally, the performance of Model 1 and Model 2 was compared. In the testing set, the accuracy, sensitivity, specificity, F1-score, and the area under the receiver operating characteristic (ROC) curve (AUC) of Model 1 reached 77.65, 86.05, 76.19, 81.42%, and 0.839, respectively. For Model 2, accuracy improved by 6.87%, sensitivity by 4.43%, specificity by 9.52%, F1-score by 7.38%, and AUC by 0.086, respectively. The models in our study could provide efficient clinical auxiliary diagnostic efficiency for fungal keratitis.

Diagnostic Efficacy of LAMP Assay for Human Fungal Pathogens: a Systematic Review and Meta-analysis.

Human fungal infections particularly caused by Candida and Aspergillus have emerged as major public health burden. Long turnaround time and poor sensitivity of the conventional diagnostics are the major impediments for faster diagnosis of human fungal pathogens. To overcome these issues, molecular-based diagnostics have been developed. They offer enhanced sensitivity but require sophisticated infrastructure, skilled manpower, and remained expensive. In that context, loop-mediated isothermal amplification (LAMP) assay represents a promising alternative that facilitates visual read outs. However, to eradicate fungal infections, all forms of fungi must be accurately detected. Thus, a need for alternative testing methodologies is imperative that should be rapid, accurate and facilitate widespread adoption. Therefore, the aim of the present study is to conduct a meta-analysis to assess the diagnostic efficiency of LAMP in the detection of a panel of human fungal pathogens following PRISMA guidelines using scientific databases viz. PubMed, Google Scholar, Science Direct, Scopus, BioRxiv, and MedRxiv. From various studies reported on the diagnosis of fungi, only 9 articles were identified as eligible to meet the criteria of LAMP based diagnosis. Through this meta-analysis, it was found that most of the studies were conducted in China and Japan with sputum and blood as the most common specimens to be used for LAMP assay. The collected data underlined that ITS gene and fluorescence-based detections ranked as the most used target and method. The pooled sensitivity values of meta-analysis ranged between 0.71 and 1.0 and forest plot and SROC (summary receiver operating characteristic) curve revealed a pooled specificity values between 0.13 and 1.0 with the confidence interval of 95%, respectively. The accuracy and precision rates of eligible studies mostly varied between 70 to 100% and 68 to 100%, respectively. A quality assessment based on QUADAS-2 (Quality Assessment of Diagnostic Accuracy Studies) of bias and applicability was conducted which depicted low risk of bias and applicability concerns. Together, LAMP technology could be considered as a feasible alternative to current diagnostics considering high fungal burden for rapid testing in low resource regions.

Modern Technologies in Diagnosis of Fungal Keratitis (Review).

Traumas and infectious diseases of the eye play a leading role in the development of corneal blindness responsible for 1.5-2 million cases of vision loss per year. To date, the issue of reducing the incidence of fungal keratitis is acute and needs to be solved worldwide. Trauma as a risk factor for corneal fungal disease is thought to be prevalent in developing countries due to agricultural involvement, while in developed countries the onset of the disease is predisposed by medical advances such as contact vision correction and modern ophthalmic surgery. Thorough analysis of the pathogenesis gives the possibility to describe the action of fungal enzymes, biofilm formation, and the resistance mechanism, which on the one hand explains the aggressive course of the disease and difficulties in its diagnosis, and on the other hand, it encourages searching for new methods of diagnosis and treatment. The non-specific clinical picture of fungal keratitis, the variety and availability of antibiotics nowadays become an obstacle for rapid detection of this pathology. Low public awareness and late visit to an ophthalmologist are also a barrier to successful combating the increasing incidence of fungal keratitis. Belated diagnosis, increasing resistance of fungi to antibiotics, and lack of registered antifungal ophthalmic drugs justify poor treatment efficacy resulting in decreased visual acuity or vision loss. Existing diagnostic methods need systematization and detailed comparison, identifying the advantages and disadvantages of each. This review considers causative agents and their influence on pathogenesis of the disease, describes difficulties of fungal keratitis diagnosis and possible ways of overcoming these problems using new developments, and also outlines further prospects of research in this direction.

Updates in Laboratory Identification of Invasive Fungal Infection in Neonates.

Invasive fungal infection (IFI) in immunocompromised neonates is significantly associated with high morbidity and mortality and has become the third most common infection in Neonatal Intensive Care Units. The early diagnosis of IFI for neonatal patients is difficult because of the lack of specific symptoms. The traditional blood culture remains the gold standard in clinical diagnosis for neonatal patients but it requires a long duration, which delays treatment initiation. Detections of fungal cell-wall components are developed for early diagnosis but the diagnostic accuracy in neonates needs to be improved. PCR-based laboratory methods, such as real-time PCR, droplet digital PCR, and the cationic conjugated polymer fluorescence resonance energy transfer (CCP-FRET) system, distinguish the infected fungal species by their specific nucleic acids and show a high sensitivity and specificity. Particularly, the CCP-FRET system, which contains a cationic conjugated polymer (CCP) fluorescent probe and pathogen-specific DNA labeled with fluorescent dyes, could identify multiple infections simultaneously. In the CCP-FRET system, the CCP and fungal DNA fragments can self-assemble into a complex with an electrostatic interaction and the CCP triggers the FRET effect under ultraviolet light to make the infection visible. Here, we summarize the recent laboratory methods for neonatal IFI identification and provide a new perspective for early clinical fungal diagnosis.

Progress of polymer-based strategies in fungal disease management: Designed for different roles.

Fungal diseases have posed a great challenge to global health, but have fewer solutions compared to bacterial and viral infections. Development and application of new treatment modalities for fungi are limited by their inherent essential properties as eukaryotes. The microorganism identification and drug sensitivity analyze are limited by their proliferation rates. Moreover, there are currently no vaccines for prevention. Polymer science and related interdisciplinary technologies have revolutionized the field of fungal disease management. To date, numerous advanced polymer-based systems have been developed for management of fungal diseases, including prevention, diagnosis, treatment and monitoring. In this review, we provide an overview of current needs and advances in polymer-based strategies against fungal diseases. We high light various treatment modalities. Delivery systems of antifungal drugs, systems based on polymers' innate antifungal activities, and photodynamic therapies each follow their own mechanisms and unique design clues. We also discuss various prevention strategies including immunization and antifungal medical devices, and further describe point-of-care testing platforms as futuristic diagnostic and monitoring tools. The broad application of polymer-based strategies for both public and personal health management is prospected and integrated systems have become a promising direction. However, there is a gap between experimental studies and clinical translation. In future, well-designed in vivo trials should be conducted to reveal the underlying mechanisms and explore the efficacy as well as biosafety of polymer-based products.

Artificial intelligence-aided rapid and accurate identification of clinical fungal infections by single-cell Raman spectroscopy.

Integrating artificial intelligence and new diagnostic platforms into routine clinical microbiology laboratory procedures has grown increasingly intriguing, holding promises of reducing turnaround time and cost and maximizing efficiency. At least one billion people are suffering from fungal infections, leading to over 1.6 million mortality every year. Despite the increasing demand for fungal diagnosis, current approaches suffer from manual bias, long cultivation time (from days to months), and low sensitivity (only 50% produce positive fungal cultures). Delayed and inaccurate treatments consequently lead to higher hospital costs, mobility and mortality rates. Here, we developed single-cell Raman spectroscopy and artificial intelligence to achieve rapid identification of infectious fungi. The classification between fungi and bacteria infections was initially achieved with 100% sensitivity and specificity using single-cell Raman spectra (SCRS). Then, we constructed a Raman dataset from clinical fungal isolates obtained from 94 patients, consisting of 115,129 SCRS. By training a classification model with an optimized clinical feedback loop, just 5 cells per patient (acquisition time 2 s per cell) made the most accurate classification. This protocol has achieved 100% accuracies for fungal identification at the species level. This protocol was transformed to assessing clinical samples of urinary tract infection, obtaining the correct diagnosis from raw sample-to-result within 1 h.

The estimated burden of fungal diseases in Mali.

Mali is a developing country facing several health challenges with a high rate of tuberculosis (TB) and a moderate HIV infection burden. Little is known or done about fungal diseases, yet they represent a significant public health problem in certain populations. The aim of this study was to estimate the national burden of fungal disease, and summarize data, diagnostic and treatment gaps. We used national demographics and PubMed searches to retrieve articles on published data on these infections and at-risk populations (pulmonary TB, HIV/AIDS patients, patients receiving critical care etc.) in Mali. The estimated Malian population was 21,251,000 in 2020 (UN), of which 45% were children <14 years. Among HIV patients, we estimate an annual incidence of 611 cryptococcosis, 1393 Pneumocystis pneumonia, 180 histoplasmosis and >5,700 esophageal candidiasis and some microsporidiosis cases. Our prevalence estimates for tinea capitis are 2.3 million, for recurrent vulvovaginal candidiasis 272,460, ∼60,000 fungal asthma and 7,290 cases of chronic pulmonary aspergillosis (often mistaken for TB). Less common acute fungal infections are probably invasive aspergillosis (n=1230), fungal keratitis (n=2820), candidaemia (>1,060) and mucormycosis (n=43). Histoplasmin was found in 6% in general population. A few cases of mycetoma are described in Mali. Many WHO Essential medicines and Diagnostics are not available in Mali. This shows a marked disparity in documented and estimated cases of fungal diseases in Mali. These infections are underestimated due to the lack of accurate diagnosis tools and lack of support for fungal diseases diagnosis and management.

Conventional and rapid diagnosis of fungal infections including mucormycosis linked to COVID-19

Abstract: Introduction: COVID-19 is a primary respiratory viral illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV2). It has been a challenge to diagnose COVID-19-associated mucormycosis as the clinical and radiological findings are nonspecific and could overlap. This study was done to estimate the incidence of various fungal infections during the COVID 19 pandemic and to evaluate the merits of conventional microbiological diagnostic approach. Method: All the nasal swabs and endoscopic biopsy samples received for fungal diagnosis from suspected mucormycosis COVID positive or COVID suspected patients were included in the study and were processed as per conventional microbiological techniques including direct KOH mount and culture growth on LPCB. Results: A total of 434 samples from suspected mucormycosis patients were received during the study period including 214 nasal swabs and nasal biopsies each, 1 sample each of nasal fluid, BAL and pus along with 3 sputum samples. The positivity rate in direct microscopy of swabs was 38.3%, and for tissue samples was 72.4%. Culture positivity rate was 47.6% amongst swabs with 19 Mucorales, 41 Aspergillus spp., 22 dematiaceious and 20 Candida spp. Culture positivity rate amongst biopsy samples was 40.6% with 35 for Mucorales, 36 for Aspergillus spp., 8 for dematiaceious and 8 for Candida spp. Conclusion: In our study, tissue samples had a greater culture positivity rate as compared to the nasal swabs, so they should be the preferred samples to be sent to the laboratory. However, nasal swabs performed better in culture growth, so their value need not be underestimated. In countries like India where RT-PCR not available in all clinical setups conventional techniques such as KOH wet mounts from samples and culture findings hold a promising outcome to start the antifungal therapy.

Applications of Reflectance Confocal Microscopy in the Diagnosis of Fungal Infections: A Systematic Review.

Cutaneous and adnexal fungal infections are typically diagnosed with potassium hydroxide (KOH) skin scrapings, fungal cultures, and Periodic acid-Schiff (PAS) biopsy staining. All three current methods of fungal diagnosis require sample processing and turnover time which leads to a delay in diagnosis. Reflectance confocal microscopy (RCM) is a non-invasive, in vivo skin imaging technology that provides real-time dermatologic diagnoses. We present an updated systematic review of the applications of RCM in diagnosing fungal infections in an effort to explore the utility of RCM as an adjunct clinical tool in detecting cutaneous and adnexal fungi We systematically searched the MEDLINE (via PubMed) for studies published from January 2000 to October 2022 that described the utility of RCM in the setting of fungal infections. Of the 25 studies that met the inclusion criteria, 202 patients were included. The following information on the application of RCM in the setting of fungal infections was extracted from each study, if reported: study type, year published, number of patients included, diagnosis/diagnostic methods, and RCM description. Concordant within all included studies, fungal infections presented on RCM as bright, linear, branching, filamentous structures at the level of stratum corneum. A limitation of this review is that 11 of 25 studies were case reports (n = 1). Larger scale studies should be conducted to explore the utility of RCM in diagnosing fungal infections and to enrich the RCM descriptions of specific fungal conditions.

Optimizing cultivation conditions for the highest expression of fungal allergens

BackgroundAny reliable allergy diagnosis depends on the quality of the testing material. In the case of fungal allergy, fungal extracts, typically used as test solutions, exhibit considerable differences in their allergenicity. Better knowledge of fungal allergen expression would enable the production of diagnostic fungal extracts of higher quality and, thus, improve the specificity and sensitivity of fungal allergy diagnosis. ObjectiveOur study aimed to find optimal cultivation conditions for the highest expression of fungal allergens. MethodsFungal species (Alternaria alternata, Ulocladium chartarum, Aspergillus fumigatus, Cladosporium herbarum, and Paecilomyces variotii) were cultivated under different conditions, and extracts were prepared from fungal material. To detect the expression of the homologous major allergens Alt a 1 and Ulo c 1 and of different fungal enolases, Western blots with allergen-specific antibodies were carried out. ResultsWestern blots performed with antibodies directed against Alt a 1 and enolases showed that the expression of fungal allergens is highly species-dependent. Even allergens of closely related fungal species and highly conserved, cross-reactive allergens display different expression patterns. ConclusionThis study exhibits the impact of different environmental conditions on the expression of the fungal allergens Alt a 1, Ulo c 1, and different fungal enolases. Furthermore, it broadens the knowledge regarding the expression pattern of the major fungal allergens Alt a 1 and Ulo c 1. Information obtained in this study will help to optimize fungal cultivation to produce diagnostic fungal extracts of high quality and, therefore, improve diagnostic specificity and sensitivity.

FcMBL magnetic bead-based MALDI-TOF MS rapidly identifies paediatric blood stream infections from positive blood cultures.

Rapid identification of potentially life-threatening blood stream infections (BSI) improves clinical outcomes, yet conventional blood culture (BC) identification methods require ~24-72 hours of liquid culture, plus 24-48 hours to generate single colonies on solid media suitable for identification by mass spectrometry (MS). Newer rapid centrifugation techniques, such as the Bruker MBT-Sepsityper® IVD, replace culturing on solid media and expedite the diagnosis of BCs but frequently demonstrate reduced sensitivity for identifying clinically significant Gram-positive bacterial or fungal infections. This study introduces a protocol that utilises the broad-range binding properties of an engineered version of mannose-binding lectin linked to the Fc portion of immunoglobulin (FcMBL) to capture and enrich pathogens combined with matrix-assisted laser desorption-ionisation time-of-flight (MALDI-TOF) MS for enhanced infection identification in BCs. The FcMBL method identified 94.1% (64 of 68) of clinical BCs processed, with a high sensitivity for both Gram-negative and Gram-positive bacteria (94.7 and 93.2%, respectively). The FcMBL method identified more patient positive BCs than the Sepsityper® (25 of 25 vs 17 of 25), notably with 100% (3/3) sensitivity for clinical candidemia, compared to only 33% (1/3) for the Sepsityper®. Additionally, during inoculation experiments, the FcMBL method demonstrated a greater sensitivity, identifying 100% (24/24) of candida to genus level and 9/24 (37.5%) top species level compared to 70.8% (17/24) to genus and 6/24 to species (25%) using the Sepsityper®. This study demonstrates that capture and enrichment of samples using magnetic FcMBL-conjugated beads is superior to rapid centrifugation methods for identification of BCs by MALDI-TOF MS. Deploying the FcMBL method therefore offers potential clinical benefits in sensitivity and reduced turnaround times for BC diagnosis compared to the standard Sepsityper® kit, especially for fungal diagnosis.

Fungal Endocarditis in Non Neutropenic Patient: A Case Report

Fungal disease of cardiac valve is rare and occurs mostly in patients with predisposing conditions. Clinical presentation is variable and could lack classic signs and symptoms of bacterial endocarditis, as well as the onset of disease vary from nonspecific signs to severe systemic embolic complications, leading to a changeling early diagnosis and higher mortality. In this case report, a 50-year-old man presenting to emergency room for stupor and fever, rapidly worsen with respiratory failure, shock and admission to intensive care unit next day. Twelve days after, transthoracic echocardiography identifies a vegetation on native mitral valve, and blood cultures positive for yeast like fungi leads to Fungal Endocarditis diagnosis.

The Case for Fungal Keratitis to Be Accepted as a Neglected Tropical Disease.

Amongst the treatable cause of blindness among young people, fungal keratitis ranks high. There are an estimated 1,051,787 to 1,480,916 eyes affected annually, with 8–11% of patients having to have the eye removed. Diagnosis requires a corneal scraping, direct microscopy and fungal culture with a large number of airborne fungi implicated. Treatment involves the intensive application of antifungal eye drops, preferably natamycin, often combined with surgery. In low-resource settings, inappropriate corticosteroid eye drops, ineffective antibacterial therapy, diagnostic delay or no diagnosis all contribute to poor ocular outcomes with blindness (unilateral or bilateral) common. Modern detailed guidelines on fungal keratitis diagnosis and management are lacking. Here, we argue that fungal keratitis should be included as a neglected tropical disease, which would facilitate greater awareness of the condition, improved diagnostic capability, and access to affordable antifungal eye medicine.

Does exposure to different menstrual products affect the vaginal environment?

The vaginal ecosystem is a key component of women's health. It also represents an ideal system for ecologists to investigate the consequence of perturbations on species diversity and emerging properties between organizational levels. Here, we study how exposure to different types of menstrual products is linked to microbial, immunological, demographic, and behavioural measurements in a cohort of young adult women who reported using more often tampons (n=107) or menstrual cups (n=31). We first found that cup users were older and smoked less than tampon users. When analysing health indicators, we detected potential associations between cups use reporting and fungal genital infection. A multivariate analysis confirmed that in our cohort, reporting using cups over tampons was associated with the higher odds ratio to report a fungal genital infection diagnosis by a medical doctor within the last 3months. We did not detect significant differences between groups in terms of their bacterial vaginal microbiota composition and found marginal differences in the level of expression of 20 cytokines. However, a multivariate analysis of these biological data identified some level of clustering based on the menstrual product type preferred (cups or tampons). These results suggest that exposure to different types of menstrual products could influence menstrual health. Larger studies and studies with a more powered setting are needed to assess the robustness of these associations and identify causal mechanisms.

Demographic and Socioeconomic Factors Associated with Fungal Infection Risk, United States, 2019.

Abstract Fungal infections cause substantial rates of illness and death. Interest in the association between demographic factors and fungal infections is increasing. We analyzed 2019 US hospital discharge data to assess factors associated with fungal infection diagnosis, including race and ethnicity and socioeconomic status. We found male patients were 1.5–3.5 times more likely to have invasive fungal infections diagnosed than were female patients. Compared with hospitalizations of non-Hispanic White patients, Black, Hispanic, and Native American patients had 1.4–5.9 times the rates of cryptococcosis, pneumocystosis, and coccidioidomycosis. Hospitalizations associated with lower-income areas had increased rates of all fungal infections, except aspergillosis. Compared with younger patients, fungal infection diagnosis rates, particularly for candidiasis, were elevated among persons >65 years of age. Our findings suggest that differences in fungal infection diagnostic rates are associated with demographic and socioeconomic factors and highlight an ongoing need for increased physician evaluation of risk for fungal infections.

Demographic, clinical, and laboratory findings of mushroom-poisoned patients in Kermanshah province, west of Iran.

BackgroundMushroom poisoning can cause gastrointestinal, renal, and hepatic symptoms and even death. This descriptive study examined the demographic, clinical, and laboratory findings of patients with fungal poisoning, a type of fungus causing the poisoning, and the incidence and mortality rates of fungal poisoning in Kermanshah province, western Iran, from 2014 to 2018.MethodsThe medical records of 193 patients with mushroom poisoning from 2014 to 2018 were evaluated. The liver and kidney function tests, electrolytes, abdominal and pelvic ultrasound, chest x-ray, coagulation tests, and coagulation factors (fibrinogen, prothrombin) were assessed. Data were collected from the medical records of patients admitted to the Poisoning Center of Imam Khomeini Hospital in Kermanshah, Iran using a researcher-made checklist. Data were analyzed by SPSS (version 16) using descriptive statistics, including mean, standard deviation, and frequency distribution tables. Trend analysis for proportion was done by chi-square statistics in STATA-14 software (ptrend command).ResultsOf cases, ‌51.3% were male, 92.6% were city dwellers, 38.3% were aged 21–40 years, and 92.5% were poisoned during the spring. The fungus that caused poisoning was Amanita virosa. The gastrointestinal, nervous, and visual systems were the most common systems involved. The most common gastrointestinal symptoms included nausea and vomiting (72.0%) and abdominal pain (71.0%). Vertigo (11.9%) and headache (9.3%) were the most common neurological symptoms. The most common visual manifestation was blurred vision (7.8%). Of cases, 23.7% had metabolic acidosis. The increased alkaline phosphatase level was the most common liver disorder in 98.7% of the cases. Increased blood urea nitrogen and creatinine levels were also reported in 21.0% and 17.7% of the cases, respectively. The serum lactic dehydrogenase and creatine phosphokinase levels also increased in 99.3% and 30.2% of the patients, respectively. The mortality rate was 1.6% (n = 3).ConclusionThe fungal poisoning diagnosis should always be considered in young patients referred to the emergency department with gastrointestinal complaints, a history of consuming wild self-picked mushrooms, and high liver and kidney test values. Since most fungal poisonings occur in the spring, it is necessary to inform the community of the dangers of consuming self-picked wild mushrooms, especially in this season.

P420 Fungal detection by means of HCR using 2D-Covalent Organic Framework Nanosheet

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMThe timely diagnosis of fungal infections is of prime importance for prescribing appropriate anti-fungal drugs. Current methods for fungal diagnosis involve culture-based methods, antibody-based detection using lateral flow assays and RT-PCR. In the present work, we devised a non-enzymatic amplification using 2D-Covalent Organic Framework (COF) nanosheet for the detection of fungal DNA.Objectives(1) Validation of exfoliated 2D COF Nanosheet as an efficient DNA detection tool via Hybridization chain reaction (HCR) triggered fluorescent assay. (2) Sequence retrieval and probe generation of fungal sample and detection of extracted target DNA via fluorescent assay.MethodA novel COF was synthesized and characterization was done using FTIR, BET, TGA, XRD, and SEM. Probes for the detection of fungi (Candida, Aspergillus, and Mucor) were designed using NUPACK software. HCR was monitored for different time and probe concentrations and standardized reaction was used for the detection of fungal RNA.ResultsFTIR, BET, TGA, XRD, and SEM confirmed the structure and formation of COF-nanosheet. H1, H2 probes at a concentration of 1μm and in presence of Target DNA (0.001μm-1μm) showed HCR reaction at 1.5 h. Fluorescence quenching was observed when probes were mixed with both, bulk COF and COF-nanosheets but increased quenchingConclusionFungal detection can be done by means of HCR using the Covalent nanosheets.

Laboratory diagnostic methods for medically important fungi

Fungi can cause a plethora of infections. They are often difficult to detect and identify. Various phenotypic and genotypic tests are there for fungal diagnosis. Here in this review article the author has tried to explain the extant culture and identification methods for various fungi of medical importance,

COVID-19 and Fungal infections: a double debacle

Fungal infections remain hardly treatable because of unstandardized diagnostic tests, limited antifungal armamentarium, and more specifically, potential toxic interactions between antifungals and immunosuppressants used during anti-inflammatory therapies, such as those set up in critically ill COVID-19 patients. Taking into account pre-existing difficulties in treating vulnerable COVID-19 patients, any co-occurrence of infectious diseases like fungal infections constitutes a double debacle for patients, healthcare experts, and the public economy. Since the first appearance of SARS-CoV-2, a significant rise in threatening fungal co-infections in COVID-19 patients has been testified in the scientific literature. Better management of fungal infections in COVID-19 patients is, therefore, a priority and requires highlighting common risk factors, relationships with immunosuppression, as well as challenges in fungal diagnosis and treatment. The present review attempts to highlight these aspects in the three most identified causative agents of fungal co-infections in COVID-19 patients: Aspergillus, Candida, and Mucorales species.