bluuebunny/crossref_metadata_2025 huggingface dataset:
P285 Predominance of Trichophyton tonsurans causing tinea capitis: a 12-years retrospective study in north of Iran
Authors: Shokohi Tahereh , Hedayati Mohammad Taghi , Kermani Firoozeh , Moosazadeh Mahmood , Nosratabadi Mohsen , Asadi Sabrieh , Salimi Maryam , Javidnia Javad , Aghili Seyed Reza
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Background

Tinea capitis is a common and endemic fungal infection of the scalp that Trichophyton spp. and Microsporum spp. usually cause. This study aimed to investigate tinea capitis and its etiologic agents for 12 years in northern Iran.

Material Methods

The study is a retrospective analysis involving 11 095 patients referred to the Reference Laboratory of Medical Mycology at Sari, Iran, from July 2009 to April 2022. The skin scraping and hair samples were assessed based on direct microscopy and culture, and causative agents were identified based on macroscopic, and microscopic morphology.

Results

Tinea capitis was confirmed in 209 patients: 157 (75.1%) male and 52 (24.9%) female out of 921 suspected patients with a scalp lesion. The prevalence of tinea capitis in patients who refer to the Reference Laboratory of Medical Mycology varied from 6.1 to 37.5%. In both sexes, a higher rate of tinea capitis was observed in patients younger than 20 years of age. Trichophyton tonsurans (146/209; 69.9%) was the most etiological agent, followed by T. mentagrophytes (13/209; 6.2), T. violaseum (9/209; 4.3), Microsporum canis (3/209; 1.4%), T. verrucosum (2/209; 1%), and T. rubrum (1/209; 0.5%). On direct microscopy examination, endothrix hair invasion in 77.0% cases, ectothrix in 2.3%, septate hyphae in 10.5%, and ecto-endothrix in one case were observed.

Conclusion

In recent decades, the prevalence of tinea capitis caused by anthropophilic fungi has remained high, particularly in younger children. Therefore, it is essential to focus on public and personal health education in this age group to prevent and control the disease.
P286 Histoplasmosis in Sri Lanka
Authors: Sigera Liyanage Shamithra Madhumali , Jayasekera Primali I , Denning David W
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Background

Histoplasmosis is a systemic mycosis caused by Histoplasma capsulatum, a dimorphic fungus. More severe disease has been observed in immunocompromised individuals. Most cases occur in certain endemic regions of the world however it seems to have a much wider global distribution. Histoplasmosis is infrequently recognized in Sri Lanka and the dispersion of information on cases is fragmented.

Method

The comprehensive search of medical literature in the English language through databases from any time to February 2022. Either, culture-proven or histopathologically proven cases were selected as diagnostically confirmed histoplasmosis. Duplicate reports were excluded. All available data on demography, clinical presentation, diagnostic method, management, and clinical outcome were appraised for the reported cases.

Result

One survey of histoplasmin skin sensitivity testing and ten cases of histoplasmosis across Sri Lanka were observed during the above period. A total of 5.7% of histoplasmin positivity had been observed in the survey of histoplasmin sensitivity among 1366 Sri Lankan volunteers of the Western and the Central Provinces in 1969. Most of the patients were reported from the Central province which had the positive histoplasmin test previously. In addition, cases were observed in Southern Province, the Sabragamuwa Province, and the Eastern Province. The majority of affected individuals were adult males (90%) and pediatric patients were not observed. The clinical presentation stretched from oral lesions (the most common presentation), skin lesions, and fever of unknown origin, to adrenal crisis. Disseminated histoplasmosis was diagnosed in 50% of the patients however asymptomatic, acute pulmonary, and chronic pulmonary histoplasmosis was not observed. Both diabetes and betel chewing are likely to be linked with oral histoplasmosis and none of the patients were positive for HIV. Both histopathology and fungal culture methods were used for the diagnosis while the use of antigen and antibody testing were not popular. Both itraconazole and amphotericin B were used for the treatment of the patients with variable outcomes.

Conclusion

Histoplasmosis exists in Sri Lanka. The number of cases could be expected to be much higher than reported along with the increase in at-risk populations. These mandates enhance laboratory diagnostic facilities and increase the awareness of medical professionals in Sri Lanka.
P287 A case of cerebral aspergillosis in Sri Lanka
Authors: Welagedara P.G.R.I.S. , Sigera Liyanage Shamithra Madhumali , Devakanthan B , Jayasekara P.I.
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Introduction

Cerebral aspergillosis is a rare disease entity that is usually associated with high mortality. In accordance with most systemic fungal infections, immunocompromised patients are the predominant victims affected. Nonspecific clinical and radiological features may contribute to difficulty and delay in diagnosis, further contributing to the poor prognosis of cerebral aspergillosis. Obtainment of proper specimens for laboratory diagnosis is a key to make a timely diagnosis that may be lifesaving.

Case Report

A 74-year-old female presented with a frontal headache for 2 weeks’ duration. She had been diagnosed with a low-grade lymphoma and had completed the chemotherapy and high-dose prednisolone 4 weeks prior to this presentation. Other than the frontal headache, which progressed slowly over 2 weeks, she did not disclose any associated symptoms. Physical examination was normal, with neither sign of raised intracranial pressure nor of a CNS infection. Basic investigations revealed no abnormalities, and the white cell count was 7.5 × 109/l and the CRP was 7 mg/l.

An MRI of the brain indicated right frontal sinusitis with a secondary small abscess in the right frontal lobe inferiorly. Therefore, she was treated with parenteral antibiotics for a bacterial infection and was discharged.

The patient presented with a headache again after 3 months. A repeat MRI of the brain revealed an increase in the size of the lesion in the right frontal lobe and the possibility of a cerebral tumor was suggested. A diagnostic biopsy was performed, and the specimen was subjected to fungal studies. The direct microscopic examination revealed fungal filaments and the culture yielded a pure growth of Aspergillus fumigatus.

She was started on intravenous amphotericin B, followed by oral voriconazole. Her headache gradually subsided with antifungal therapy. The duration of therapy was guided by serial radiological imaging, and the patient achieved a complete recovery at the end of 1-year of treatment. She remains asymptomatic to date, after 2 years of treatment completion.

Discussion

Immunocompromised patients with cerebral aspergillosis may present with minimal clinical symptoms and signs. Obtaining a proper specimen for laboratory testing is vital to arrive at a definitive diagnosis. Radiological investigations may play an important role in the diagnosis as well as during the follow-up of a patient with cerebral aspergillosis. Appropriate antifungal treatment for a prolonged duration, with or without neurosurgical intervention and reversal or reduction of immunosuppressive therapy leads to a good prognosis.
P294 A descriptive study of Trichosporon fungemia cases from tertiary care center from north India
Authors: Srinivasan Priya , Kaur Harsimran , Gupta Parakriti , Pandey Abhishek , Paul Raees A , Ghosh Anup , Chakrabarti Arunaloke , Rudramurthy Shivaprakash M
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Objective

Incidence of fungaemia with Trichosporon spp is increasing especially in immunocompromised patients. High mortality of 60% is associated with T. fungemia. Trichosporon species are intrinsically resistant to echinocandins, exhibit high MICs to amphotericin B, and have a propensity to form biofilm necessitating accurate and timely diagnosis. Cases of Trichosporon fungemia are often misdiagnosed and under-reported owing to the difficulties in diagnosis. Here, we report the clinical presentation and outcome of Trichosporon fungemia cases at the tertiary care center from north India.

Methods

This is a descriptive study conducted at the Department of Medical Microbiology, PGIMER, Chandigarh, India. A total of 8 patients who had fungemia due to Trichosporon spp. were included in the study. Trichosporon spp. was identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) with an updated database. Antifungal susceptibility was done using microbroth dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI). Beta-D-glucan (BDG) assay was performed as per the manufacturer's recommendation. Demographic and clinical data along with treatment outcomes of all the patients were noted from medical records.

Results

The average age of presentation of patients diagnosed with Trichosporon fungemia was 44 years and the male to female ratio was 5:3. The underlying conditions included necrotizing pancreatitis (n = 2), infective endocarditis (n = 2) and renal transplant (n = 1) The other risk factors included the use of broad-spectrum antibiotics (n = 7), a central venous catheter (n = 4), and prior surgical procedures (n = 3) BDG was tested in 5 patients and 4 patients had a positive value of >80 pg/ml. A total of 5/8 patients received antifungal treatment. Six patients clinically improved and were discharged while two patients died of refractory shock despite treatment with amphotericin B. Minimum inhibitory concentration of Trichosporon isolates was (range): amphotericin B 0.5-16 μg/ml, fluconazole 0.12-32 μg/ml, voriconazole 0.03-1 μg/ml, itraconazole 0.03-1 μg/ml, and posaconazole 0.03-0.5 μg/ml.

Conclusion

Trichosporon spp. is an opportunistic pathogen causing fungemia in immunocompromised patients. Most of the patients in our study were immunocompetent except for one post-transplant patient. BDG helps in the diagnosis of this infection. This study highlights the need to accurately diagnose Trichosporon infections and perform antifungal susceptibility testing for guiding appropriate management and reducing mortality.
P300 Importance of direct microscopy in respiratory mold infection: a diagnostic dilemma
Authors: Tilak Ragini , Gupta Munesh Kumar , Narwade Prity , Bhatia Mohit
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Introduction

Opportunistic fungal infections are on the rise due to a changing healthcare paradigm worldwide. India is the second most populated country in the world and is experiencing an upward trend in fungal lung diseases.

Objective

Rapid diagnosis of respiratory mold infections by direct microscopy.

Material and Methods

We retrospectively analyzed the mycology of the received respiratory samples; sputum, BAL, or ETT secretion between January 1, 2021 to March 31, 2022. All the samples were processed as per standard mycological procedures. For each sample, two KOH wet mounts were prepared and examined. Samples were inoculated over SDA and blood agar and incubated at 28°0C and 37°C in BOD. Once there was growth, it was processed as per standard protocol.

Results

We received a total of 109 respiratory samples from suspected respiratory fungal infections of which early morning sputum samples 84 (77%) were the predominant followed by BAL samples 25 (23%). Most of the enrolled patients were male 73(67%) with a mean age of 48 years. Among 109 samples, 83 (76%) were shown to be fungal elements by KOH wet mount while 26 (24%) were KOH negative. A total of 53 (48.6%) samples were culture-positive Aspergillus flavus30(27%), was found to be the most common species isolated followed by A. fumigatus 16 (14.6%), miscellaneous11(10%), Aspergillus. species 5 (4.5%), and A. terreus 2 (1.8%).

Conclusion

Considering the findings of KOH mount in our study, we would like to emphasize that this is a very simple, rapid, and cost-effective laboratory method with high diagnostic sensitivity when correlated with clinical findings, radiological findings, and host factors.

Further, it has been found to be very dependable for making decisions to start empirical antifungal therapy early by clinicians.
P304 Epidemiological and clinical characteristics of patients with onychomycosis caused due to Neoscytalidium spp. in two reference centers in Bogotá D.C.
Authors: Álvarez María Alejandra Vanegas , Hernández Oscar Mora , Prada Leticia Sopó , Riveros John Hernández , Osorio Elvis Gómez
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Objective

To describe the sociodemographic and clinical characteristics, diagnostic methods, and therapies received in patients with onychomycosis confirmed due to Neoscytalidium spp, who attended the San José Hospital and the Medical Mycology Laboratory for 1 year, in order to get a better perspective of the disease in our field, enhance its detection and increase early prevention.

Methods

A descriptive cross-sectional observational study was carried out in two centers: San José Hospital and Medical Mycology Laboratory from November 2019 to November 2020; patients of all ages with a confirmed diagnosis of onychomycosis due to Neoscytalidium spp. were included. The cases were identified based on ICD-10 codes and the information was collected from the clinical history, questionnaires, clinical photographs, and additional documents.

Results

A total of 465 cases of onychomycosis were confirmed in the chosen period of time, of which 60 cases were caused due to Neoscytalidium spp. Affecting 31 women and 29 men, with an average age of 48.5 years in both sexes; 85% of these patients lived in Bogota D. C. The mean duration between the onset of the disease and its diagnosis was 3 years, 13% of the patients had a history of chronic disease including thyroid disorders, arterial hypertension, and diabetes mellitus. A total of 98% of the patients had toenail involvement, 88% had 1 to 5 nails involved, 33% of the cases showed mild nail involvement, 20% moderate, and 6% severe. Likewise, 35% of the cases presented dermatomycosis. 43% of the patients reported having received topic and systemic treatment, where there was no improvement in the major of the cases (78%).

Conclusions

Our results support what is reported in the literature regarding risk factors, sociodemographic findings, and difficulty in therapeutic response to the pharmacological measures available to treat this infection; as well as the opinion of certain authors, onychomycosis caused due to non-dermatophyte mold is clinically indistinguishable from that caused by dermatophytes. Therefore, a complete mycological study (direct examination/KOH and culture) should always be performed to identify the causative agent before starting any treatment, since as our study showed, infections due to Neoscytalidium spp. are, in general frequency, the third most common cause of infection in onychomycosis and the first by non-dermatophyte molds.
P341 Evaluation of anti-Candida activity of Cinnamaldehyde using mice model
Authors: Uttam Gunjan , Singh Karuna
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objective

Vaginal candidiasis is a frequent infection afflicting the female population. Candida co-infections are reported in escalating frequency and can be associated with severe health hazards or even death. People with severe COVID-19, malignancies, AIDS, and organ transplants are particularly vulnerable to invasive vaginal candidiasis. Due to the high frequency of infections associated with recurrence, vaginal candidiasis poses a significant medical problem worldwide. Treatments of vaginal candidiasis are limited due to drug resistance, side effects, and toxicity. CIN is a natural compound and its antifungal activity is widely reported. The introduction of cinnamaldehyde (CIN) as the anti-Candidal agent will revolutionize the treatment of vaginal candidiasis.

In this study, we investigated the anti-Candida activity of CIN against vaginal candidiasis in Swiss albino mice (C3HHC-Strain).

Methods

Vaginal candidiasis in mice (Swiss albino) was induced under conditions of pseudo-estrus. Persistent vaginal infection was found in estrogenized mice after vaginal challenge with C. albicans. The mice were treated orally after confirmation of infection in mice. The efficacy of CIN treatment was investigated phenotypically by colony-forming unit (CFU) counts in the vaginal smear, fungal load determination in the blood, the ovarian and vaginal tissues and periodic acid-Schiff (PAS) staining of histopathological sections of the vaginal tissues. The hematological parameters of the experimental mice were also evaluated.

Results

The pseudohyphae and spores of C. albicans were present in the vaginal smear of experimentally infected mice. After treatment, no C. albicans colonies were found in the vaginal lavage of infected mice. The fungal burden was significantly higher in the vagina and the ovaries of infected mice. However, a dose of 262.5 mg/kg BW of CIN reduced the number of CFU in the vagina and ovaries in treated mice. The histopathology revealed the absence of C. albicans in the vaginal tissue of the treated mice. Nonetheless, the vaginal sections of infected mice exhibited pathological changes. The hematological parameters such as RBC count, WBC count, and percentage of hemoglobin showed significant differences in the treatment groups compared to the infected group.

Conclusion

Cinnamaldehyde showed good in vivo antifungal potential against vaginal candidiasis. However, evaluation of its pharmacodynamic and pharmacokinetic parameters and complete elucidation of its mode of action are desirous.
P349 Relationship between genotypes and in vitro pathogenic behavior of Uruguayan strains of Cryptococcus spp.
Authors: Cabeza Elisa , Arteta Zaida , Betancor Laura
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objective

Contribute to the knowledge of the relationship between genetic and pathogenic characteristics of meningeal cryptococcosis-producing strains from Uruguay.

Methods

We have included a total of 21 isolates of Cryptococcus spp from human cases of meningoencephalitis. We start from the general strain collection of the mycology laboratory of the department of parasitology and mycology, medical school, University of the Republic. The strain was sectioned taking into account the phenotypic diversity and genetic diversity through a previous analysis by PCR fingerprinting, for study by MLST.

MLST was performed according to the protocol available at: http://mlst.mycologylab.org/defaultinfo.aspx?Page=CryptoHome.

Infection, intracellular survival, and cell proliferation assays were performed. The murine macrophage cell line J774 (ATCC TIB 67) was used.

Results

Of the 21 strains studied, in 13 we obtained an ST that had been previously reported (9 C. neoformans complex and 4 C. gattii complex). For the remaining 8 strains, we obtained new allelic combinations to which new STs were assigned, which were deposited in the MLST database for Cryptococcus spp. (http://mlst.mycologylab.org) (Table 1).

All strains were phagocytosed and all were able to persist within macrophages at 24 h post-infection, although there was great variability between strains (Table 1). In most of the strains, the control of macrophages prevailed through cell lysis mechanisms over Cryptococcus spp, with one proliferation index (PI) ˂1.0. In the proliferation assay, only 4 strains managed to proliferate within the macrophages, 3 of them correspond to C. gattii complex, genotypes VGI and VGII. The 4th strain that achieved proliferation was corresponding to the VNI genotype.

Conclusion

We obtained a clear differentiation of in vitro behavior between the strains corresponding to C. neoformans complex and C. gattii complex. The strains belonging to C. gattii complex presented less phagocytosis and greater persistence and intracellular proliferation.
P384 Characterization of the virulence potential of Aspergillus species of section Terrei in Galleria mellonella infection model
Authors: Shahandashti Roya Vahedi , Binder Ulrike , Houbraken Jos , Lass-Flörl Cornelia
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objectives

Species belonging to the genus of Aspergillus are among the most common causative agents of human and animal infections. Less than 40 species among all Aspergillus species are known to be associated with human infections, including allergic bronchopulmonary aspergillosis, chronic pulmonary aspergillosis, and invasive aspergillosis. And of these, Aspergillus section Fumigati is one of the major infectious causes of death, followed by members of sections Flavi, Nigri, and Terrei. Aspergillus species in section Terrei are categorized into three series: Ambigui, Nivei, and Terrei. A. terreus sensu stricto is the first species described and the most common species found worldwide in different ecological habitats.

However, there are several other species within the section Terrei, stating cryptic species which are not distinguished by conventional morphological analysis, even though they are taxonomically accepted by forming a distinctive phylogenetic clade. Despite definitive species identification, there is still less known about the virulence potential of all species in this section, and it might be underestimated because of their lack of distinction by conventional diagnostic methods. In this ongoing study, the in vivo Galleria mellonella model has been utilized to examine the inter-and/or intraspecies virulence dependency of section Terrei.

Methods

A total of 18 accepted Aspergillus species in section Terrei (n = 18) were tested, including A. terreus sensu stricto, A. citrinoterreus, A. hortae, A. pseudoterreus, A. alabamensis, A. aureoterreus, A. floccosus, A. iranicus, A. recifensis, A. carneus, A. microcysticus, A. niveus, A. bicephalus, A. neoindicus, A. neoafricanus, A. barbosae, A. ambiguus, and A. allahabadii. Species were identified by sequencing gene regions of b-tubulin, calmodulin, and RNA Polymerase II Subunit 2 (RPB2). Briefly, groups of larvae (n = 30) (0.3 to 0.4 g; SAGIP, Italy) were stored in wood shavings in the dark at 18°C for 24 h before the experiment. Three groups were included: larvae infected with 107 conidia/larva, larvae injected with 20 μL sterile insect physiological saline, and untouched larvae. The survival rate was monitored for up to 144 h at 37°C.

Results

Median survival rates revealed a species-dependent virulence pattern. Larvae inoculated with A. aureoterreus, A. pseudoterreus (Serie Terrei) and A. niveus, A. carneus, and A. iranicus (Serie Nivei) exhibited high virulence potential by reflecting lower survival rates in comparison with other species. In contrast, species belonging to the series Ambigui showed low virulence potential.

Conclusion

In conclusion, the virulence characteristics of section Terrei differ between species. Further studies are needed to unravel the species' invasiveness, such as histopathology and immune response of G. mellonella.
P396 Utility of nasal swabs as samples for the diagnosis of Mucormycosis using molecular methods for detection
Authors: Chowdhury Souradeep , Sikdar Sunit , Ranjan Piyush , Singh Gagandeep , Pandey Mragnayani , Ghosh Tamoghna , Sachdev Janya , Yadav Renu , Perumalla Sowjanya , Bhattacharya Aakashneel , Xess Immaculata , Wig Naveet
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objectives

Mucormycosis is an aggressive, life-threatening infection caused by fungi in the order Mucorales. There was an explosion of new cases of rhino-sino-orbital mucormycosis following the COVID pandemic in India, and the need for easy and rapid diagnostics was felt. The current diagnosis of mucormycosis relies on mycological cultures, radiology, and histopathology. These methods lack sensitivity and are most definitive later in the course of infection, resulting in the failure of timely intervention.

A real-time multiplex PCR platform is commercially available for the detection of Rhizopus spp., Mucor spp. Rhizomucor spp., Lichtheimia spp., and Cunninghamella spp. (PN-700, MucorGenius®, PathoNostics®, Maastricht, The Netherlands) This real-time PCR has been validated to identify these fungal pathogens from bronchoalveolar lavage, tissue, and serum samples. This study aimed to validate this PCR-based system to detect Mucorales from nasal swab samples and evaluate its utility in the detection of Mucorales from nasal cavities of high-risk patients developing signs and symptoms of mucormycosis.

Methods

A single-center cross-sectional observational study was conducted on 50 hospitalized adult patients with signs and symptoms of mucormycosis. Nasal swabs were taken for PCR analysis once there was a clinical suspicion and were compared with the results of the gold standard. The gold standard for the diagnosis of mucormycosis was the conventional method (KOH mounted microscopy/HPE). Demographic details and risk factors for these patients were recorded, and the RTPCR-based test was run on the nasal swab samples of all these 50 patients. The workflow is depicted graphically in Fig. 1 (Created with BioRender.com).

Results

The study population mean (SD) age was 50 (16) years and consisted of 32 (64%) males. A total of 39 (78%) patients were known cases of diabetes mellitus, 48 (96%) patients had amphotericin B intake, and 20 (40%) had posaconazole intake. In all, 21 (42%) patients had a past history of COVID-19 infection; 14 patients had received steroids and 10 patients received oxygen support. PCR for Mucorales was positive in 15 (30%) patients while the KOH mount was positive in 4 (8%) patients.

Conclusion

These results are not encouraging for the use of nasal swabs as the sample for diagnosis of mucormycosis. Though the PCR performed better on the swab samples than KOH preparation and culture techniques, it highlights the importance of using standard sampling methods.
P4 COVID19-associated fungal infections
Authors: Hoenigl Martin
Published: September 2022

Plenary session 4, September 22, 2022, 2:00 PM - 2:45 PM

 

 

Coronavirus disease 2019 (COVID-2019) associated invasive fungal infections (IFIs) have emerged as an important complication in a substantial number of critically ill COVID-19 patients, and three groups of fungal pathogens have increasingly been recognized as causes of superinfections: Aspergillus, Mucorales, and Candida. First reports of cases and case series of COVID-19-associated pulmonary aspergillosis (CAPA) have emerged during the first months of the pandemic. Prevalence rates varied widely due to the fact that CAPA was, and still remains, challenging to diagnose in patients with COVID-19-associated acute respiratory failure (ARF). The clinical picture and radiological findings of CAPA are unspecific and the primarily airway invasive growth in non-neutropenic patients and the late occurrence of angioinvasion in the course of the disease may complicate diagnosis. Current guidelines recommend treatment of CAPA during its early airway invasive phase, which may result in some overtreatment (i.e., treatment in patients that may not develop angioinvasive infection), given the independent contribution of CAPA to devastating mortality rates of around 50% that have been shown in multiple studies. This talk will also review the incidence of COVID-19-associated IFIs caused by Mucorales, and Candida, including C. auris, and will discuss—clinical risk factors, predisposing changes in the host environment, and immunological mechanisms involved in the pathogenesis of these coinfections, as well as current and future diagnostics and treatments.
P402 Comparison of PCR-RFLP with 21-plex PCR and rDNA: sequencing for identification of clinical yeast isolates
Authors: Khodavaisy Sadegh , Kord Mohammad , Elmimoghaddam Ahmad , Hashemi Seyed Jamal , Reziae Sassan , Ghazvini Roshanak Daie , Salehi Mohammadreza , Abdollahi Alireza , Ahmadi Ali , Boekhout Teun
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objectives

Non-albicans Candida species and other rare yeasts have emerged as major opportunistic pathogens in fungal infections. Identification of opportunistic yeasts in developing countries is mainly performed by phenotypic assay, which is time-consuming and prone to errors. The aim of the present study was to evaluate PCR-RFLP as a routinely used identification technique for the most clinically important Candida species in Iran and make a comparison with a novel multiplex PCR, called 21-plex PCR.

Methods

A total of 173 yeast isolates from clinical sources were selected and identified with sequence analysis of the D1/D2 domains of rDNA (LSU rDNA) sequencing as the gold standard method.

Results

The results were compared with those obtained by PCR-RFLP using MspI restriction enzyme and the 21-plex PCR. PCR-RFLP correctly identified 93.4% of common pathogenic Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and P. kudriavsevii (=C. krusei)) and was able to identify 45.5% of isolates of the uncommon yeast species compared to the D1/D2 rDNA sequencing. Compared with PCR RFLP, all common Candida species and 72.7% of uncommon yeast species were correctly identified by the 21-plex PCR.

Conclusion

The application of the 21-plex PCR assay as a non-sequence-based molecular method for the identification of common and rare yeasts can reduce turnaround time and costs for the identification of clinically important yeasts and can be applied in resource-limited settings.
P403 Metagenomic sequencing as an effective diagnostic tool for Talaromycosis in HIV-negative patients: A retrospective study
Authors: Liang Tianwei , Cao Cunwei
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objective

The diagnosis of Talaromyces marneffei (T. marneffei) infection in HIV-negative patients is still challenging. The aim of this study was to evaluate the effectiveness and efficiency of metagenomic next-generation sequencing (mNGS) for T. marneffei infection in non-HIV-infected patients by comparing the diagnostic value of mNGS and traditional microbiological culture on different specimens.

Methods

A total of 57 HIV-negative patients with suspected infectious diseases were enrolled in the study from July 2018 to November 2021. mNGS and culture were performed on all enrolled samples. Data were collected on demographic characteristics, clinical symptoms, laboratory findings, and antifungal results. The diagnostic value of mNGS and conventional microbiological culture was compared.

Results

A total of 66 samples from 57 patients were analyzed, 29 were diagnosed with T. marneffei infection and 28 were of non-T. marneffei infected patients. Compared with the final diagnosis, mNGS showed a sensitivity of 97.22%, which was higher than that of conventional culture (61.11%). In addition, mNGS provides greater diversity in sample selection compared to culture.

Conclusion

Our study demonstrated that mNGS can be considered a promising tool for rapid and accurate pathogenic diagnosis in HIV-negative patients with suspected T. marneffei.
P423 Recombinase polymerase amplification for Mucormycosis
Authors: Hallur Vinaykumar , Sable Mukund , C. Preetam , Sahu Supriya , Tudu Malati
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objectives

(1) To develop a Recombinase polymerase amplification (RPA)-based assay to diagnose mucormycosis. (2) To determine the analytical sensitivity and cross-reactivity of the developed assay. (3) To validate the assay in clinical samples. (4) To determine the turnaround time for the developed assay.

Methods

Multiple alignments of the ITS region of Rhizopus arrhizus were used to identify conserved sequences and design 30-32 bp long primers and 45 bp long Exo probes for RPA-based assay as per TwistDX guidelines (Cambridge, UK) using the PrimedRPA program (Higgins et al. 2019) and subject to in silico specificity check using NCBI primer blast. Genomic DNA and plasmid clone of the target region were spiked into human DNA and were used to determine analytical sensitivity. DNA from 35 clinically relevant fungi and bacteria were used to determine the cross-reactivity. A total of 40 sinus tissue samples from patients with microscopy and culture-confirmed mucormycosis and 40 sinus tissue samples from patients without mucormycosis were used to determine the diagnostic sensitivity and specificity of the assay. DNA from relevant fungi, bacteria and molds were extracted using the MPbio FastDNA kit as per the manufacturer’s instructions with an elution volume of 100 μl. All extracted DNA samples were subjected to GAPDH PCR before RPA testing to rule out the presence of inhibitors as per established protocols. A 50 μl RPA reaction using Twistamp Exo kit as per manufacturer's instruction but using 10 μl template instead of 1 μl. The reactions were set up and inserted into a portable Qiagen ESE quant TS2.4 fluorometer incubated at 39°C and fluorescence was acquired every 30 sec over 20 min. The tubes were removed from the fluorometer mixed well and reinserted after 4 min as per kit instructions. Positive control and no template control were set up with each reaction. All relevant data were entered in Microsoft excel and diagnostic sensitivity and specificity were calculated using the Medcalc free online statistical software.

Results

A total of 1246 primer-probe combinations were designed and checked for in silico specificity using NCBI primer blast. Of these 1246 primer probes, 8 primers and 1 probe was finalized for in vitro testing to determine the best primer and probe set. The finalized primers had analytical sensitivity of 10pg and 10 copies and showed no cross-reactivity with any of the 35 clinically relevant fungi and bacteria. All extracted DNA showed amplified GAPDH gene demonstrating the absence of PCR inhibitors in extracted DNA. The sensitivity and specificity of the assay were 97.5% (95% CI: 86.8-99.9) and 97.6% (95% CI: 87.1-99.9). The turnaround time for RPA was 6 h.

Conclusion

Real-time RPA can be used to rapidly and reliably diagnose mucormycosis.
P443 Altered expression of fungal CotH, human glucose-regulated protein 78 (GRP78), and predicted miRNAs in macrophages and model diabetic mice infected with Rhizopus oryzae
Authors: Shokohi Tahereh , Seifi Zahra , Shafie Mohammad , Mowla Javad , Niknejad Farhad , Abastabar Mahdi , Larijan Lale Vahedi
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objectives

Rhizopus oryzae is one of the most common causes of mucormycosis. Among the virulence factors of the Mucorales, CotH protein has recently been identified, which causes the invasion of R. oryzae into endothelial cells. In this study, we aimed to examine the reaction between GRP78 at the level of human cells and different groups of mice and CotH3 at the surface of the R. oryzae hyphae. We evaluated the relative expression of GRP78 and CotH3 genes and changes in the expression of some miRNAs that target the human GRP78 gene.

Methods

In this study, the relative changes in gene expression were studied. In three groups (1) Macrophages derived from human monocytes: monocytes from the blood of healthy donors were isolated using Ficoll and in RPMI 1640 medium containing FCS 10% and with penicillin-streptomycin after 2 weeks were differentiated into macrophages. Two groups were investigated, including control and infected with R. oryzae hyphae, for 6 and 16 hours after infection. (2) Hematogenous dissemination mucormycosis model: Seven groups of male BALb/c mice were examined in control, infected, and treated groups with Liposomal amphotericin B. (3) Human mucormycosis: in this study, two samples of patients with rhinocerebral mucormycosis with diabetes mellitus treated and untreated with liposomal amphotericin B were examined. Total RNA extraction and cDNA synthesis were performed from the studied samples. The relative expression changes of the target genes and miRNAs were evaluated using real-time PCR carried out using Sybrgreen-based detection methods.

Results

Monocyte-derived macrophages had a steady pattern in relative changes in gene expression. An increase in expression of two genes, GRP78 and CotH3, was observed in the samples, and all miRNAs targeted by the GRP78 gene included hsa - miR-16-5p; hsa -miR-335-5p and hsa -miR-93-3p showed a decreasing pattern. In the mice mucormycosis model, relative gene expression changes were observed, and mmu-miR-181b-5p showed increased expression deviation in all groups. The clinical sample of diabetic patients with untreated rhinocerebral mucormycosis also had a consistent pattern of GRP78 and CotH3 increased gene expression. The hsa-miR-16-5p and hsa-miR-335-5p have increased expression, while hsa-miR-93-3p decreased expression.

Conclusion

After validation, these micro RNAs can be used as valuable markers in mucormycosis detection and treatment processes.
P446 Molecular detection of fungal agents responsible for COVID-19-associated mycosis directly from tissue specimens
Authors: Singh Vikramjeet , Das Anupam , Agarwal Ashish Chandra , Gupta Nikhil , Sen Manodeep , Agarwal Jyotsna
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objective

Due to the ongoing COVID-19 pandemic a new group of patients at risk emerged with COVID-19-associated mucormycosis (CAM) and other fungal infections. Molecular studies, evaluating the prevalence of CAM and other fungal infections are lacking. To assess CAM prevalence in a super-specialty healthcare hospital in North India, we applied direct microscopy, fungal culture, and qualitative real-time-PCR targeting Mucorales-specific fragments on tissue specimens of critically ill COVID-19 patients.

Methods

This was a hospital-based prospective study during second-wave of COVID-19 in India. All clinically suspected CAM patients with a history of COVID-19 were included in the study from March 2021 to June 2021 where tissue or biopsy specimens were collected under aseptic conditions. Conventional identification methods were performed for all isolates, speciation was done by MALDI-TOF, and comparative detection by RTPCR was also done.

Results

In the present study, among 67 samples received in the laboratory from clinically suspected CAM patients, 32 samples showed positive growth using the conventional method of identification. Rhizopus arrhizus was the commonest fungal isolate obtained followed by Aspergillus flavus from tissue samples. Use of molecular and automated machines helped in the early identification of these species 24-48 h less than the conventional methods. Polyfungal isolates are also reported from two tissue samples of patients in the post-COVID discharge stage. Almost 90% of patients with CAM and other fungal etiology agreed to steroid intake and diabetes condition during COVID-19 infection.

Conclusion

Considering the ever-evolving strains and variants of COVID-19, it is important to have a high index of suspicion for fungal coinfection in patients with COVID-19 presenting with comorbidities. Further, they should undergo immediate molecular studies with an emphasis on the requirement of medical or surgical intervention if the result comes positive. There is a need to stress on the judicious use of steroids to avoid flaring up of the fungal infection.
P459 The Aspergillus lateral flow assay for the diagnosis of chronic pulmonary aspergillosis
Authors: Peng Jie , Yan Jun , Zhang Yuan , Yao Wang , Wang He , Su Yan , Zhou Zeqi
Published: September 2022

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

 

Objectives

Chronic pulmonary aspergillosis (CPA) is an uncommon and problematic pulmonary disease, complicating many other respiratory disorders. Measurement of Aspergillus-specific IgG antibodies had a higher sensitivity than either IgM, IgA, or IgE and it should therefore be considered the most appropriate test for screening CPA. Aspergillus-specific IgG antibody levels have been successfully used to monitor the response of CPA to medical therapy. Recently, a novel rapid test for Aspergillus-specific IgG antibody from Dynamiker Biotechnology (Tianjin) Co., Ltd. was released as a screening test of CPA. It is a fluorescent immunochromatographic cassette test using a monoclonal antibody against Aspergillus-specific IgG antibody and Europium nanoparticles (Eu NP) (Fig. 1). It is a semi-quantitative test which was easy to operate, rapid with portable detection devices, and can be widely accepted by clinical and primary medical (Fig. 2). We collect clinical samples to verify the detection performance of the Dynamiker Quic™Aspergillus-specific IgG antibody (LFA) test.

Methods

In total, 102 patients were included and 42 patients were diagnosed with CPA. The pathogen was identified from sputum, BALF culture, lung resection surgery, bronchos copy biopsy, percutaneous lung biopsy and BALF GM assay.

Results

The sensitivity, specificity, PLR, NLP and Kappa index of the IgG antibody test were 85.6%,94.4%,14.0%,0.23% and 0.72, respectively.

Conclusions

The current work indicates that the Dynamiker Quic™ Aspergillus specific IgG antibody (LFA) test shows a promising application in the diagnosis of CPA. The results are accurate and reliable, and it could be used as an aid for the early rapid screening test of CPA.
P473 Genomic epidemiology of antifungal-resistant Candida auris in Colombia
Authors: Escandon Patricia , Misas Elizabeth , Gade Lalitha , Caceres Diego , Hurst Steve , Litvintseva Anastasia , Duarte Carolina , Chow Nancy A.
Published: September 2022

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

 

Introduction

Candida auris is a public health threat. Five major clades of C. auris have been identified (Clades I–V). In Colombia, C. auris infections were first reported in 2016 with ongoing transmission reported from multiple cities. Here, we describe C. auris genomic epidemiology in Colombia detailing cases from 2016–2021.

Methods

A total of 99 isolates from C. auris cases were collected between June 2016 to January 2021 in Colombia, representing 11 geographic locations. Species confirmation, antifungal susceptibility testing, and whole-genome sequencing (WGS) were performed. In all, 37 genomic sequences generated previously from isolates from C. auris cases in Colombia, Venezuela, Panama, Israel, and United States were also analyzed MycoSNP workflow was used to assess sequence quality, map reads to the reference, and identify single-nucleotide polymorphisms (SNPs). Pairwise distances and a neighbor-joining tree were generated. IQtree was used to generate a maximum-likelihood tree with bootstrap values.

Results

Phylogenetic analysis identified 1 493 SNP positions. Isolates from Colombia clustered to Clade IV and predominately grouped by country except for 16 fluconazole-resistant isolates from Bogota, Colombia that grouped with five isolates from Venezuela. In this cluster, 20 (95%) were resistant to fluconazole and 5 (24%) were resistant to fluconazole and the echinocandin micafungin. Remaining isolates from Bogota did not group in this cluster and were susceptible to fluconazole and micafungin.

A total of 98 isolates from Colombia clustered together. Within this Colombian cluster, there were two subgroups that had bootstrap support of 100% and were separated by 13 SNPs. The first subgroup was a cluster that contained 18 isolates from the north coast; 17 (94%) isolates were resistant to amphotericin B. A second subgroup consisted of 26 isolates from Cesar and Norte de Santander, and 22 (84%) isolates were resistant to fluconazole.

Conclusions

Based on the phylogenetic reconstruction, C. auris in Colombia continues to be of Clade IV. Amphotericin B-resistant isolates were predominately from the north coast, fluconazole-resistant isolates were from a wider geographic area in Colombia, and echinocandin-resistant isolates were from Bogota. Within the Colombian cluster comprising two subgroups, we observed high genetic relatedness between isolates from different geographic locations suggesting transmission among cities.
P474 The value of PCR-based azole resistance detection in invasive aspergillosis: A prospective multicenter study
Authors: Huygens Sammy , Dunbar Albert , Buijl Jochem , Maertens Johan , Verweij Paul , Klaassen Corne , Lagrou Katrien , Van Dijk Karin , Mercier Toine , Schauwvlieghe Alexander , Rijnders Bart
Published: September 2022

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

 

Objectives

Prompt detection of azole-resistant Aspergillus fumigatus will result in the timely start of active treatment and may improve the survival of invasive aspergillosis (IA). The use of a multiplex polymerase chain reaction (PCR) targeting Aspergillus species and fumigatus DNA as well as the two most prevalent azole resistance- associated mutations (RAMs) in the cyp51A gen (TR34/L98H and TR46/Y121F/T289A) could shorten the time to detect azole-resistant IA.

Methods

In a prospective study in 12 Dutch and Belgian centers, we evaluated the clinical value of the multiplex AsperGenius®PCR in hematology patients with a pulmonary infiltrate undergoing bronchoalveolar lavage (BALf) sampling. The primary endpoint was antifungal treatment failure in the 6 weeks after antifungal treatment initiation in the patients in which azole-resistant IA was detected. Treatment failure was defined as death or a switch to an antifungal agent from another class after at least 5 days of first-line therapy. Patients with a mixed azole-susceptible/resistant infection were excluded from this analysis to ascertain that the infection was indeed caused by the resistant strain.

Results

Of 323 patients enrolled, sufficient BALf for PCR testing remained in 299. Probable fungal disease was diagnosed in 95 (34%), Aspergillus cultured in 24 (8%), Aspergillus DNA detected in 118 (39%), and A. fumigatus DNA in 88 (29%) patients. The resistance PCR was conclusive in 54/88 (61%) and RAMs were detected in 8 (15%), Table 1. All 8 had probable IA but 2 had a mixed infection and were excluded. In the 6 remaining patients, treatment failure was observed in one. Compared with the GM negative patients and despite antifungal therapy, a positive GM test was associated with a 13% higher 6-week overall mortality (P = .01), Table 2. Surprisingly, the 6-week mortality in the 65 patients who had a positive Aspergillus PCR but a negative GM and culture was not increased compared to those with a negative PCR (PCR + 14% vs. PCR- 16% mortality, P = .68).

Conclusions

In patients with an underlying hematological disease and a pulmonary infiltrate, the detection of Aspergillus DNA by PCR on BALf was not associated with increased mortality. The exact place of the Aspergillus PCR in the EORTC-MSGERC invasive fungal infection criteria is therefore uncertain. In 15% of the patients in whom A. fumigatus DNA was present, azole RAMs were detected by PCR. In only 1/6 probable cases of IA with RAMs detected, antifungal treatment failure was observed. Basing the choice of antifungal therapy on the result of a cyp51a resistance PCR may help to reduce the impact of azole resistance on mortality.