Molecular and microscopy detection of Pneumocystis jirovecii in hospitalized patients during the COVID-19 pandemic.

Introduction: Early detection of Pneumocystis jirovecii as an opportunistic pathogen that may endanger predisposed persons, including COVID-19 patients, may help to choose the optimal management. Methods: In this study, 585, including 530 COVID-19 patients, with clinical and radiological evidence of respiratory diseases, were investigated for P. jirovecii screening. Clinical specimens were examined by direct microscopy and PCR, and randomly selected positive PCR products were confirmed through DNA sequence analysis. Results: Thirty-one (5.3%) samples were positive in P. jirovecii-specific nested-PCR, while by direct microscopic tests, Pneumocystis was observed in 22 (3.76%) samples. Males (61.7%) and patients over 50 years old (75.6%) were more commonly affected than others, and malaise and fatigue (84%), and wheezing (75%) were the most common symptoms, followed by fever (40.48%) and dyspnea (39.51%). Among the Pneumocystis-positive patients, three cases had coinfection with Aspergillus fumigatus, A. flavus, and A. niger (each n = 1), as documented by direct microscopy, culture, and species identification by PCR-sequencing. Conclusion: Pneumocystis pneumonia is still a diagnostic challenge; therefore, additional large-scale studies are needed to clarify the epidemiology of the disease in immunocompromised or COVID-19 patients.

Efficacy of Hydroxychloroquine in Pre-exposure Severe Acute Respiratory Syndrome Coronavirus 2 Prophylaxis among High-Risk HealthCare Workers: A Multicenter Study

Background: Prophylaxis could be an established strategy to potentially prevent and control infectious diseases and should be considered in the coronavirus disease 2019 (COVID-19) pandemic. The present study aimed to assess the effectiveness of hydroxychloroquine as a prophylaxis treatment strategy in the reduction of the risk of COVID-19 among health professionals. Materials and Methods: The health professionals were randomly assigned (1:1) to the control group without receiving any hydroxychloroquine as prophylaxis and the hydroxychloroquine group receiving a weekly hydroxychloroquine dose of 400 mg up to 12 weeks. Results: A total of 146 health professionals were randomly enrolled in this study between August 11 and November 11 in 2020. Among the screened health professionals, 21 (14.6%) were infected with COVID-19 during the 12 weeks, and 14 (66.6%) out of the 21 health professionals were in the control group. Most participants with COVID-19 had mild symptoms (62%). In addition, 9.5% (n = 2) of the participants suffered from moderate disease and 28.5% were diagnosed with severe symptoms. In the hydroxychloroquine group, 5 (7.1%) and 2 (2.8%) participants were reported with mild and moderate symptoms of COVID-19, respectively, and 2 participants had moderate, 8 (10.9%) participants had mild symptoms, and 6 (8.2%) participants had severe symptoms in the control group, within 3 months. Severe symptoms of COVID-19 were not observed in the hydroxychloroquine group. Conclusion: This study addressed the effect and benefit of hydroxychloroquine administration for the prevention of COVID-19 among health professionals. The improved perception of prophylaxis might highlight its important role in future COVID-19 outbreaks to prevent hospital transmission, which is a major route of spread.

The Seroprevalence and Seropositivity of SARS-CoV-2 among Healthcare Workers during the Third Pandemic Wave.

BACKGROUND: Due to the unclear protective role of produced antibodies and the need for seroepidemiologic studies, we surveyed the COVID-19 seroprevalence among healthcare professionals who had direct or indirect contact with COVID-19 patients. METHODS: From 19 October 2020 to 17 February 2021, 300 healthcare workers were enrolled and tested for serum antibodies in this prospective cohort study. Demographic information, risk factors, and infection history were collected. Anti- SARS-CoV-2 IgG and IgM antibody titers were determined to estimate the seroconversion rate. RESULTS: During the first and second phases of the study, the positive seroconversion rates were 31.7 and 26.6%, respectively. In seronegative individuals, sixteen (10.6%) new cases of COVID-19 and five (6.3%) reinfections were identified. Among those with a positive antibody level, forty-one (36.9%) healthcare workers reported no symptoms in the preceding months. There was no association between occupational exposure and an increased probability of seroconversion. CONCLUSIONS: The seropositivity rate and the rate of asymptomatic individuals with seroconversion was remarkable and could be an indicator of a high infection rate among healthcare workers.

Efficacy of Hydroxychloroquine in Pre-exposure Severe Acute Respiratory Syndrome Coronavirus 2 Prophylaxis among High-Risk HealthCare Workers: A Multicenter Study.

Background: Prophylaxis could be an established strategy to potentially prevent and control infectious diseases and should be considered in the coronavirus disease 2019 (COVID-19) pandemic. The present study aimed to assess the effectiveness of hydroxychloroquine as a prophylaxis treatment strategy in the reduction of the risk of COVID-19 among health professionals. Materials and Methods: The health professionals were randomly assigned (1:1) to the control group without receiving any hydroxychloroquine as prophylaxis and the hydroxychloroquine group receiving a weekly hydroxychloroquine dose of 400 mg up to 12 weeks. Results: A total of 146 health professionals were randomly enrolled in this study between August 11 and November 11 in 2020. Among the screened health professionals, 21 (14.6%) were infected with COVID-19 during the 12 weeks, and 14 (66.6%) out of the 21 health professionals were in the control group. Most participants with COVID-19 had mild symptoms (62%). In addition, 9.5% (n = 2) of the participants suffered from moderate disease and 28.5% were diagnosed with severe symptoms. In the hydroxychloroquine group, 5 (7.1%) and 2 (2.8%) participants were reported with mild and moderate symptoms of COVID-19, respectively, and 2 participants had moderate, 8 (10.9%) participants had mild symptoms, and 6 (8.2%) participants had severe symptoms in the control group, within 3 months. Severe symptoms of COVID-19 were not observed in the hydroxychloroquine group. Conclusion: This study addressed the effect and benefit of hydroxychloroquine administration for the prevention of COVID-19 among health professionals. The improved perception of prophylaxis might highlight its important role in future COVID-19 outbreaks to prevent hospital transmission, which is a major route of spread.

CT findings of COVID-19-associated pulmonary aspergillosis: a systematic review and individual patient data analysis.

PURPOSE: Common CT abnormalities of pulmonary aspergillosis represent a cavity with air-meniscus sign, nodule, mass, and consolidation having an angio-invasive pattern. This study aims to conduct a systematic review and an individual patient-level image analysis of CT findings of COVID-19-associated pulmonary aspergillosis (CAPA). METHODS: A systematic literature search was conducted to identify studies reporting CT findings of CAPA as of January 7, 2021. We summarized study-level clinical and CT findings of CAPA and collected individual patient CT images by inviting corresponding authors. The CT findings were categorized into four groups: group 1, typical appearance of COVID-19; group 2, indeterminate appearance of COVID-19; group 3, atypical for COVID-19 without cavities; and group 4, atypical for COVID-19 with cavities. In group 2, cases had only minor discrepant findings including solid nodules, isolated airspace consolidation with negligible ground-glass opacities, centrilobular micronodules, bronchial abnormalities, and cavities. RESULTS: The literature search identified 89 patients from 25 studies, and we collected CT images from 35 CAPA patients (mean age 62.4 ± 14.6 years; 21 men): group 1, thirteen patients (37.1%); group 2, eight patients (22.9%); group 3, six patients (17.1%); and group 4, eight patients (22.9%). Eight of the 14 patients (57.1%) with an atypical appearance had bronchial abnormalities, whereas only one (7.1%) had an angio-invasive fungal pattern. In the study-level analysis, cavities were reported in 12 of 54 patients (22.2%). CONCLUSION: CAPA can frequently manifest as COVID-19 pneumonia without common CT abnormalities of pulmonary aspergillosis. If abnormalities exist on CT images, CAPA may frequently accompany bronchial abnormalities.

Melting Curve-based Assay as an Alternative Technique for the Accurate Detection of SARS-CoV-2.

Background: Early and cost-effective diagnosis and monitoring of the infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are critically important to anticipate and control the disease. We aimed to set up a SYBR Green-based one-step real-time polymerase chain reaction (PCR) as a lower-cost alternative method to detect the virus. Materials and Methods: An in-house SYBR Green-based PCR assay targeting the envelope (E) and RNA-dependent RNA polymerase (RdRp) genes, was set up to diagnose the infection, and was compared with the reference probe-based PCR method. Results: When the commercial probe-based assay was considered as the reference method, SYBR Green-based PCR had a slightly lower sensitivity (81.98% and 86.25% for E and RdRp targets, respectively) and a good specificity (100% and 94.44% for E and RdRp targets, respectively). For both gene targets, three different melting temperature (Tm) patterns were found in the PCRs of the nasopharyngeal/oropharyngeal swab samples, but no size polymorphism was seen in agarose gel electrophoresis. Conclusion: Further studies to improvement of the assay are needed to make it an inexpensive and reliable tool for the diagnosis of COVID-19.

A Chronic Autochthonous Fifth Clade Case of Candida auris Otomycosis in Iran.

Candida auris, a multidrug-resistant nosocomial pathogen, has emerged globally with high morbidity and mortality among immunocompromised individuals and COVID19 hospitalized patients. Five major clades of C. auris have been previously described. The fifth clade is exclusively found in Iran where C. auris isolates are genetically distinct from other clades by > 200,000 single-nucleotide polymorphisms. The origin of C. auris remains unclear, and limited clinical data are available at present regarding clade V infection or colonization. Herein, another case of otomycosis in Iran caused by an isolate of C. auris belonging to the fifth clade is reported. Genotyping revealed that the obtained C. auris isolate from Isfahan clustered with earlier clade V isolates from Babol, cities around 600 km separated, which indicates that C. auris clade V is established in Iran. C. auris is thought to exist more commonly in Iran, given that limited diagnostic capacity in the country has probably curbed the identification of more C. auris cases. Therefore, surveillance of the environment, patients and healthcare facilities in different geographical regions in Iran is urgently required.

Fatal disseminated aspergillosis in an immunocompetent patient with COVID-19 due to Aspergillus ochraceus.

Aspergillus infection is a well-known complication of severe influenza and severe acute respiratory syndrome coronavirus (SARS-CoV), and these infections have been related with significant morbidity and mortality even when appropriately diagnosed and treated. Recent studies have indicated that SARS-CoV-2 might increase the risk of invasive pulmonary aspergillosis (IPA). Here, we report the first case of Aspergillus ochraceus in a SARS-CoV-2 positive immunocompetent patient, which is complicated by pulmonary and brain infections. Proven IPA is supported by the positive Galactomannan test, culture-positive, and histopathological evidence. The patient did not respond to voriconazole, and liposomal amphotericin B was added to his anti-fungal regimen. Further studies are needed to evaluate the prevalence of IPA in immunocompetent patients infected with SARS-CoV-2. Consequently, testing for the incidence of Aspergillus species in lower respiratory secretions and Galactomannan test of COVID-19 patients with appropriate therapy and targeted anti-fungal therapy based on the primary clinical suspicion of IPA are highly recommended.

Fatal Invasive Pulmonary Aspergillosis in COVID-19 Patient with Acute Myeloid Leukemia in Iran.

Although patients with severe immunodeficiency and hematological malignancies has been considered at highest risk for invasive fungal infection, patients with severe pneumonia due to influenza, and severe acute respiratory syndrome coronavirus (SARS-CoV) are also at a higher risk of developing invasive pulmonary aspergillosis (IPA). Recently, reports of IPA have also emerged among SARS-CoV-2 infected patients admitted to intensive care units (ICUs). Here, we report a fatal case of probable IPA in an acute myeloid leukemia patient co-infected with SARS-CoV-2 and complicated by acute respiratory distress syndrome (ARDS). Probable IPA is supported by multiple pulmonary nodules with ground glass opacities which indicate halo sign and positive serum galactomannan results. Screening studies are needed to evaluate the prevalence of IPA in immunocompromised patients infected with SARS-CoV-2. Consequently, testing for the presence of Aspergillus in lower respiratory secretions and galactomannan in consecutive serum samples of COVID-19 patients with timely and targeted antifungal therapy based on early clinical suspicion of IPA are highly recommended.