Predictors of Intensive Care Unit Admission among Hospitalized COVID-19 Patients in a Large University Hospital in Tehran, Iran.

BACKGROUND: The rapid increase in the spread of COVID-19 and the numbers of infected patients worldwide has highlighted the need for intensive care unit (ICU) beds and more advanced therapy. This need is more urgent in resource-constrained settings. The present study aimed to identify the predictors of ICU admission among hospitalized COVID-19 patients. STUDY DESIGN: The current study was conducted based on a retrospective cohort design. . METHODS: The participants included 665 definite cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)  hospitalized in Imam Hossein Hospital from February 20 to May 14, 2020. The baseline characteristics of patients were assessed, and multivariate logistic regression analysis was utilized to determine the significant odds ratio (OR) for ICU admission. RESULTS: Participants were aged 59.52±16.72 years, and the majority (55.6%) of them were male. Compared to non-ICU patients (n=547), the ICU patients (n=118) were older, had more baseline comorbidities, and presented more often with dyspnea, convulsion, loss of consciousness, tachycardia, tachypnea, and hypoxia, and less often with myalgia. Significant OR (95% CI) of ICU admission was observed for the 60-80 age group (2.42, 95%CI: 1.01; 5.79), ≥80 age group (3.73, 95%CI: 1.44; 9.42), ≥3 comorbidities (2.07, 95%CI: 1.31; 3.80), loss of consciousness (6.70, 95%CI: 2.94, 15.24), tachypnea (1.79, 95%CI: 1.03, 3.11), and SpO2<90 (5.83, 95%CI: 2.74; 12.4). Abnormal laboratory results were more common among ICU-admitted patients; in this regard, leukocytosis (4.45, 95%CI: 1.49, 13.31), lymphopenia (2.39, 95%CI: 1.30; 4.39), elevated creatine phosphokinase (CPK) (1.99, 95%CI: 1.04; 3.83), and increased aspartate aminotransferase (AST) (2.25, 95%CI: 1.18-4.30) had a significant OR of ICU admission. Chest computer tomography (CT) revealed that consolidation (1.82, 95%CI: 1.02, 3.24), pleural effusion (3.19, 95%CI: 1.71, 5.95), and crazy paving pattern (8.36, 95%CI: 1.92, 36.48) had a significant OR of ICU admission. CONCLUSION: As evidenced by the obtained results, the predictors of ICU admission were identified among epidemiological characteristics, presenting symptoms and signs, laboratory tests, and chest CT findings.

Study of prevalence, distribution and clinical significance of Blastocystis isolated from two medical centers in Iran.

AIM: This study aimed to survey prevalence and clinical significance of Blastocystis among symptomatic and asymptomatic groups. BACKGROUND: Blastocystis is a prevalent microorganism that is found in intestine of human and majority of animals. However, most studies have failed to establish correlation between the presence of the parasite and clinical manifestations. METHODS: from Dec 2016 to Jun 2017, 554 stool samples were collected from symptomatic and asymptomatic subjects referred to Imam Hossein Hospital and Gastroenterology and Liver Diseases Research Institute, Tehran, Iran. All samples were concentrated using conventional formalin-ethyl acetate concentration and then were microscopically examined using Lugol's iodine staining and light microscope. The fresh stool samples were also cultivated in DMEM medium and were examined for growth of Blastocystis every 48 hours with direct smear slides for 10 days. RESULTS: Blastocystis was observed among 93 (16.8%) of stool samples cultivated in DMEM. The findings represented that 64/398 (16.08%) and 29/156 (18.58%) of asymptomatic and symptomatic patients were infected with Blastocystis, respectively. In addition, there was no significant correlation between presence of symptoms and carrying Blastocystis (P=0.528), although statistically significant association was observed between presence of urticaria and carrying Blastocystis (P<0.05). Furthermore, a statistically significant correlation between observing the parasite and different age groups was seen (P<0.05). CONCLUSION: Blastocystis is a prevalent parasitic eukaryote among symptomatic and asymptomatic populations despite the higher prevalence among symptomatic group that suggests the chance of infection with Blastocystis raises with age.

SYBR green-based detection of Leishmania infantum DNA using peripheral blood samples.

Parasitological methods for the diagnosis of visceral leishmaniasis (VL) require invasive sampling procedures. The aim of this study was to detect Leishmania infantum (L. infantum) DNA by real time-PCR method in peripheral blood of symptomatic VL patient and compared its performance with nested PCR, an established molecular method with very high diagnostic indices. 47 parasitologically confirmed VL patients diagnosed by direct agglutination test (DAT > 3200), bone marrow aspiration and presented characteristic clinical features (fever, hepatosplenomegaly, and anemia) and 40 controls (non-endemic healthy control-30, Malaria-2, Toxoplasma gondii-2, Mycobacterium tuberculosis-2, HBV-1, HCV-1, HSV-1 and CMV-1) were enrolled in this study. SYBR-green based real time-PCR and nested PCR was performed to amplify the Kinetoplast DNA minicircle gene using the DNA extracted from Buffy coat. From among 47 patients, 45 (95.7 %) were positive by both nested-PCR and real time-PCR. These results indicate that real time-PCR was not only as sensitive as a nested-PCR assay for detection of Leishmania kDNA in clinical sample, but also more rapid. The advantage of real time-PCR based methods over nested-PCR is simple to perform, more faster in which nested-PCR requires post-PCR processing and reducing contamination risk.

Challenging loop-mediated isothermal amplification (LAMP) technique for molecular detection of Toxoplasma gondii.

OBJECTIVE: To compare analytical sensitivity and specificity of a newly described DNA amplification technique, LAMP and nested PCR assay targeting the RE and B1 genes for the detection of Toxoplasma gondii (T. gondii) DNA. METHODS: The analytical sensitivity of LAMP and nested-PCR was obtained against10-fold serial dilutions of T. gondii DNA ranging from 1 ng to 0.01 fg. DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays. RESULTS: After testing LAMP and nested-PCR in duplicate, the detection limit of RE-LAMP, B1-LAMP, RE-nested PCR and B1-nested PCR assays was one fg, 100 fg, 1 pg and 10 pg of T. gondii DNA respectively. All the LAMP assays and nested PCRs were 100% specific. The RE-LAMP assay revealed the most sensitivity for the detection of T. gondii DNA. CONCLUSIONS: The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T. gondii. Furthermore, these findings indicate that primers based on the RE are more suitable than those based on the B1 gene. However, the B1-LAMP assay has potential as a diagnostic tool for detection of T. gondii.

Emergence of co-infection of visceral leishmaniasis in HIV-positive patients in northeast Iran: a preliminary study.

Visceral leishmaniasis (VL) serosurvey was carried out on 49 HIV/AIDS patients among 500 asymptomatic HIV/infected patients who registered in the Khorasan Razavi Province during the last 14 years. HIV infections were detected by ELISA and confirmed using western blot assay at the AIDS centre of the Khorasan Razavi Province. All collected sera were screened using the direct agglutination test (DAT). The sera with anti-Leishmania infantum antibodies at a titre of 1:100 were considered positive for VL infection and serum titration was performed from 1:100 to 1:102,400. Nine (18.4%) patients were sero-positive according to DAT. The distribution of sera titrations were as follows: 1:100 (n = 6) 1:1600 (n = 1); 1:25,600 (n = 1) and 1:102,400 (n = 1). All sero-positive cases showed clinical signs and symptoms. The most predominant signs and symptoms of co-infection of visceral leishmaniasis in HIV-positive patients were pneumonia (n = 2), hepatosplenomegaly (n = 2), lymphadenopathy (n = 2), anaemia (n = 1), prolonged fever (n = 1) and cachexia (n = 1). Our finding shows that VL (or kala-azar) is an opportunistic disease in HIV-positive patients that may be occurred in VL endemic areas of Iran.

Development and Assessment of Loop-Mediated Isothermal Amplification (LAMP) Assay for the Diagnosis of Human Visceral Leishmaniasis in Iran.

BACKGROUND: Parasitological methods for the diagnosis of Visceral leishmaniasis (VL) require invasive procedures, so serological and molecular approaches have been developed but are not generally applicable in the field. We evaluated a loop mediated isothermal amplification (LAMP) assay using blood from VL patients and compared it to nested PCR. METHODS: Forty-seven subjects with clinical features (fever, hepatosplenomegaly and anemia) were confirmed positive for VL by the direct agglutination test (DAT) at titers >3200. Forty DAT negative individuals from non-endemic areas with no clinical signs or symptoms of VL served as controls. A LAMP assay was performed using a set of six primers targeting Leishmania infantum kinetoplast DNA (kDNA) minicircle gene under isothermal (64 °C) conditions. For nested PCR we used primers targeting the kDNA minicircle gene. RESULTS: The LAMP assay provided a detection limit of 1 parasite in 1 ml of peripheral blood and detected L. infantum DNA in 44 of 47 DAT-confirmed VL cases, with diagnostic sensitivity of 93.6% (95% CI). No L. infantum DNA was amplified in controls, indicating a specificity of 100%. The nested PCR yielded sensitivity of 96% (95% CI) and a specificity of 100% (95% CI). CONCLUSION: The LAMP assay gave results similar to those of nested PCR but in a shorter time. The LAMP method is simple; requires no sophisticated equipment; has a short reaction time; and results, indicated by turbidity of the reaction mixture, are observable with the naked eye.

Cutaneous and post kala-azar dermal leishmaniasis caused by Leishmania infantum in endemic areas of visceral leishmaniasis, northwestern Iran 2002-2011: a case series.

Visceral leishmaniasis (VL) is endemic in Northwest and southern Iran. Reports of cutaneous leishmaniasis (CL) in Northwest areas are rare, and its etiological agents are unknown. In the current study, we report six CL and two post kala-azar dermal leishmaniasis (PKDL) cases caused by Leishmania infantum from endemic areas of VL in the Northwest. Smears were made from skin lesions of 30 suspected patients in 2002-2011, and CL was determined by microscopy or culture. Leishmania spp. were identified by nested-PCR assay. The disease was confirmed in 20 out of 30 (66%) suspected patients by parasitological examinations. L. infantum was identified in eight and Leishmania major in 12 CL cases by nested-PCR. Cutaneous leishmaniasis patients infected with L. major had the history of travel to CL endemic areas. L. infantum antibodies were detected by direct agglutination test (DAT) at titers of 1:3200 in two cases with history of VL. Results of this study indicated that L. infantum is a causative agent of CL as well as PKDL in the VL endemic areas.