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The Comparative Analysis of Two RT-qPCR Kits for Detecting SARS-CoV-2 Reveals a Higher Risk of False-Negative Diagnosis in Samples with High Quantification Cycles for Viral and Internal Genes.

The early detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using the real-time quantitative polymerase chain reaction (RT-qPCR) as a gold-standard molecular tool has allowed to test and trace the viral spread and the isolation of COVID-19-infected patients. The detection capacity of viral and internal genes is an essential parameter to consider and analyze during the assay. In this study, we analyze the performance of the two commercial RT-qPCR kits used in Chile, TaqMan™ 2019-nCoV Control Kit v1 (Thermo Fisher) and MaxCov19 (TAAG Genetics), for the COVID-19 diagnosis

Plastic Bronchitis of Human Bocavirus 1 Detected by Comprehensive Polymerase Chain Reaction of Mucus Casts.

Plastic bronchitis (PB) is a rare and severe respiratory disease characterized by the formation of branching mucus casts, resulting in airway obstruction. PB can be divided into two types. Type 1 PB is primarily caused by inflammatory casts that result from allergic diseases. In type 2 PB, mucinous casts are produced in association with congenital heart disease. PB is also associated with viral respiratory infections, particularly the influenza A (H1N1) pdm09 virus, which is the most common pathogen affecting pediatric patients. Herein, we report a case of severe type 1 PB caused by human boca

Assessing rodents as carriers of pathogenic Leptospira species in the U.S. Virgin Islands and their risk to animal and public health.

Leptospirosis is a global zoonotic disease caused by pathogenic bacteria of the genus Leptospira. We sought to determine if rodents in U.S. Virgin Islands (USVI) are carriers of Leptospira. In total, 140 rodents were sampled, including 112 Mus musculus and 28 Rattus rattus. A positive carrier status was identified for 64/140 (45.7%); 49 (35.0%) were positive by dark-field microscopy, 60 (42.9%) by culture, 63 (45.0%) by fluorescent antibody testing, and 61 (43.6%) by real-time polymerase chain reaction (rtPCR). Molecular typing indicated that 48 isolates were L. borgpetersenii and 3 were L. ki…

Infectivity of Symptomatic Malaria Patients to Anopheles farauti Colony Mosquitoes in Papua New Guinea.

Plasmodium transmission from humans to mosquitoes is an understudied bottleneck in the transmission of malaria. Direct membrane feeding assays (DMFA) allow detailed malaria transmission studies from humans to mosquitoes. Especially for Plasmodium vivax, which cannot be cultured long-term under laboratory conditions, implementation of DMFAs requires proximity to P. vivax endemic areas. In this study, we investigated the infectivity of symptomatic Plasmodium infections to Anopheles farauti colony mosquitoes in Papua New Guinea (PNG). A total of 182 DMFAs were performed with venous blood collecte

Peripheral blood hsa-circRNA5333-4: A novel biomarker for myasthenia gravis.

In this study, the potential of specific Circular RNAs (circRNAs) as novel peripheral blood biomarkers for myasthenia gravis (MG) was explored. We analyzed circRNAs in the peripheral blood of three normal controls and three MG patients using RNA microarray. Candidate circRNAs were validated in three independent cohorts by Quantitative Real-time polymerase chain reaction (qPCR). Eleven differentially expressed circRNAs were initially identified and four were confirmed in the first independent cohort. Hsa_circ_0076490 and hsa-circ_5333-4 had the largest areas under the curve (AUCs) of the receiv…

miR-21 regulates osteogenic and adipogenic differentiation of BMSCs by targeting PTEN.

ObjectiveTo explore the effects and mechanism of miR-21 on the osteogenic/adipogenic differentiation of mouse BMSCs.MethodsThe bilateral ovaries of C57BL/6J mice (n=24) were removed to construct an osteoporosis model. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-21, osteogenic/adipogenic genes, and PTEN. ALP and ARS and ORO staining were used to detect the formation of calcium nodules and lipid droplets in BMSCs. Western blot was used to detect the expression of PTEN.ResultsmiR-21 was significantly down-regulated in osteoporotic mice. The

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… Quantitative real-time polymerase chain reaction (qRT-PCR) The quantitative PCR enzyme mixture kit (ABI, 4366596, USA) and ABI 7900 (ABI, USA) real-time fluorescence quantitative PCR apparatus were used for detection. qRT-PCR was carried out in the 95°C environment for 30 minutes, with a succession …

Angiogenesis' related genetic variants alter clinical features and prognosis of diffuse large B-cell lymphoma patients.

OBJECTIVES: Single nucleotide variants (SNVs) in vascular endothelial growth factor A (VEGFA) and VEGFA receptor (KDR) genes confer differen … study. Patients were homogeneously treated with R-CHOP. Genotypes were identified in genomic DNA by real-time polymerase chain reaction. RESULTS: Patients with VEGFA -634CC and +936CT or TT genotypes were at increased risk of showing grade III / IV toxicities and not achieving complete remission with treatment, and shorter event-free and overall survival were seen in patients with …

Clinical course, radiological findings and late outcome in preterm infant with suspected vertical transmission born to a mother with severe COVID-19 pneumonia: a case report.

BACKGROUND: Vertical transmission of coronavirus disease 2019 (COVID-19) from mother to newborn infant is doubtful, and very little is known about disease severity and neonatal outcome. CASE PRESENTATION: We present a preterm Iranian infant born to a Persian mother with severe COVID-19 pneumonia. The mother underwent cesarean delivery, and amniotic fluid yielded a positive result for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by real-time reverse transcription polymerase chain reaction (RT-PCR). The newborn infant showed early-onset infection with SARS-CoV-2 confirmed on pha

Prevalence and clinical characteristics of septicemia in children with Mycoplasma pneumoniae pneumonia.

BACKGROUND: Mycoplasma pneumoniae (MP) pneumonia in children can be challenging to treat, and the impact of MP blood infection is unclear. The present study aims to determine the prevalence and clinical characteristics of MP septicemia among pediatric patients. METHODS: Children hospitalized at our center for MP pneumonia between October 2017 and June 2018 were included. Healthy controls visiting our outpatient clinic for regular physical examinations were also enrolled. MP was detected by real-time polymerase chain reaction (qPCR) analysis of plasma and peripheral blood mononuclear cell (PBM…

Expression of mRNA vascular endothelial growth factor in hypospadias patients.

BACKGROUND: Hypospadias is a relatively common genital anomaly in humans, usually followed by inelastic dartos that causes penile chordee. Vascular endothelial growth factor (VEGF) is strongly linked to the viscoelasticity of tissues and their elastic phase. This study aimed to evaluate VEGF expressions in (1) fascia dartos between hypospadias and controls and (2) chordee severity. METHODS: This prospective cohort study involved 65 specimens from patients with hypospadias and ten specimens from controls. The samples were analyzed by quantitative real-time polymerase chain reaction (qPCR) for …

Diagnosis of COVID-19 Based on Symptomatic Analysis of Hospital Healthcare Workers in Belgium: Observational Study in a Large Belgian Tertiary Care Center During Early COVID-19 Outbreak.

OBJECTIVE: To identify early symptoms allowing rapid appraisal of infection with SARS-CoV-2 among healthcare workers of a large Belgian hospital. METHODS: Healthcare workers with mild symptoms of an acute respiratory tract infection were systematically screened on clinical characteristics of corona virus disease 2019 (COVID-19). A nasopharyngeal swab was taken and analyzed by real-time Reverse-Transcription-Polymerase-Chain-Reaction (rRT-PCR). RESULTS: Fifty percent of 373 workers tested COVID-19 positive. The symptoms cough (82%), headache (78%), myalgia (70%), loss of smell or taste (40%),

A Rare Pediatric Case of Severe Rhabdomyolysis Owing to Dual Infection.

AIM: We aimed to report a severe and rare pediatric rhabdomyolysis case associated with a dual viral infection. CASE: A 13 year-old, healthy girl presented with the complaints of fever, abdominal pain, weakness and dark-colored urine. She was diagnosed with rhabdomyolysis based on clinical signs and laboratory findings. The diagnosis was confirmed by serological tests and real-time polymerase chain reaction for Epstein-Barr virus (EBV) and cytomegalovirus (CMV), respectively. Other potential genetic, metabolic and infectious causes were evaluated meticulously but no evidence was found. This c…

Fatal visceral disseminated varicella-zoster virus infection in a renal transplant recipient: A case report.

BACKGROUND: Visceral disseminated varicella-zoster virus (VZV) infection is a rare but life-threatening disease. In transplant recipients wi … ney injury. Next-generation sequencing of peripheral blood yielded 39224 sequence reads of VZV, and real-time polymerase chain reaction for VZV was positive, with 1.2 × 107 copies/mL. The final diagnosis was visceral disseminated VZV infection. Acyclovir and supportive therapy were started, but the patient died of severe visceral organ damage 16 h after admission. CONCLUSION: Visce …

Validation of SARS CoV-2 detection by real-time PCR in matched pooled and deconvoluted clinical samples before and after nucleic acid extraction: a study in tertiary care hospital of North India.

The diagnosis of coronavirus disease-19 (COVID-19) relies on the detection of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) RNA by real-time reverse-transcription polymerase chain reaction in respiratory samples. Rapid increase in the COVID-19 cases across the world requires fast and efficient testing as testing capacity is a bottleneck in diagnosis. In this context, pooling strategy can be opted for rapid testing in a cost-effective manner. In this study, the authors have optimized and compared the effect of pooling (5 and 10 samples) before and after nucleic acid extraction. I

Dacron swab and PBS are acceptable alternatives to flocked swab and viral transport media for SARS-CoV-2.

Nasopharyngeal flocked swabs placed in viral transport media (VTM) are the preferred collection methodology for respiratory virus testing. Due to the rapid depletion of available reagents and swabs, we have validated an alternative swab placed in phosphate-buffered saline (PBS) for use in respiratory virus testing in a SARS-CoV-2 real-time polymerase chain reaction assay and a multiplexed respiratory virus panel. We collected nasopharyngeal (NP) swabs and oropharyngeal (OP) swabs from 10 healthy volunteers. Flocked swabs were placed in VTM and alternative swabs in PBS. In this feasibility stud…

Ovarian Cancer Detection Using Photoacoustic Flow Cytometry.

Many studies suggest that the enumeration of circulating tumor cells (CTCs) may show promise as a prognostic tool for ovarian cancer. Current strategies for the detection of CTCs include flow cytometry, microfluidic devices, and real-time polymerase chain reaction (RT-PCR). Despite recent advances, methods for the detection of early ovarian cancer metastasis still lack the sensitivity and specificity required for clinical translation. Here, a novel method is presented for the detection of ovarian circulating tumor cells by photoacoustic flow cytometry (PAFC) utilizing a custom three dimensiona…

Discontinuation of tyrosine kinase inhibitors in chronic myeloid leukemia: when and for whom?

Treatment discontinuation is considered one of the main goals of therapy for patients with chronic myeloid leukemia. Several criteria are felt to be necessary to consider discontinuation, while others may predict a better chance of achieving treatment-free remission. Criteria for discontinuation include patients in chronic phase chronic myeloid leukemia, a minimum duration of tyrosine kinase inhibitor therapy of 3 years, sustained deep molecular response for at least 2 years and a molecular response of at least MR4. In addition, proper education of the patient on the need for more frequent mon

Peroxisome Proliferator-Activated Receptor Beta/Delta Agonist Suppresses Inflammation and Promotes Neovascularization.

The effects of peroxisome proliferator-activated receptor (PPAR)β/δ ophthalmic solution were investigated in a rat corneal alkali burn model. After alkali injury, GW501516 (PPARβ/δ agonist) or vehicle ophthalmic solution was topically instilled onto the rat's cornea twice a day until day 7. Pathological findings were evaluated, and real-time reverse transcription polymerase chain reaction was performed. GW501516 strongly suppressed infiltration of neutrophils and pan-macrophages, and reduced the mRNA expression of interleukin-6, interleukin-1β, tumor necrosis factor alpha, and nuclear factor-k

Oncogenic hsa_circ_0091581 promotes the malignancy of HCC cell through blocking miR-526b from degrading c-MYC mRNA.

Circular RNAs (circRNAs) are new types of endogenous non-coding RNAs, which are identified to have critical regulatory roles in cancer biology. In this study, we aimed to find the abnormally expressed circRNAs in hepatocellular carcinoma (HCC) and investigate the function and mechanism of circRNAs in HCC progression. Upregulation of hsa_circ_0091581 was identified by RNA-sequencing and validated by quantitative real-time polymerase chain reaction (qRT-PCR). Hsa_circ_0091581 expression was correlated with tumor size, disease-free survial and overall survival of HCC patients. Functionally, hsa_c…

Respiratory distress in a patient with Klinefelter syndrome: a suspicion of COVID-19 hiding severe pulmonary embolism.

Klinefelter syndrome is the most common congenital abnormality causing primary hypogonadism and predisposing to a state of hypercoagulability. We report the case of a 37-year-old man, of Algerian nationality, diagnosed with Klinefelter syndrome admitted to the hospital via the emergency room for acute chest pain and dyspnea. The patient arrived in Tunisia 36 hours ago. On admission, body temperature was 38.2°C, blood pressure, pulse and respiratory rate were 130/70 mmHg, 120/minute and 26/minute, respectively. He had an oxygen saturation of 87% in room air. His electrocardiography revealed a c