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A real-time reverse transcriptase polymerase chain reaction (RT-PCR) is the gold standard in diagnosis for infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but the false-negative result is the problem in the prevention and control the pandemic of coronavirus disease 2019 (COVID-19). A false-negative of RT-PCR test needs to be evaluated when the patient showed a high clinical suspicion for COVID-19. We report a 36-year-old man with 4 times negative RT-PCR results, but clinical, radiological (chest X-ray and chest CT scan), and serological examinations showed a high suspicion of COVID-19. History of close contacted with COVID-19 confirmed patient was reported, and the wife of our case was also RT-PCR tested positive for SARS-CoV-2 in the next few days strengthen the COVID-19 diagnosis of our case patient. It is important to use the combination of RT-PCR, chest X-ray, chest CT scan, clinical manifestations, antibodies test, and exposure history of patients to diagnose COVID-19 and decide the early isolation and appropriate treatment.

The coronavirus disease 2019 (COVID-19) outbreak started in December 2019 and rapidly spread around the globe as a major health threat. Several reports on re-positive cases subsequent to discharge from hospitals caught our attention. We aimed to highlight RT-qPCR positivity re-detection after discharge from isolation, with special consideration of the possible reasons behind it. We found that re-positive RT-qPCR assays for severe acute respiratory syndrome coronavirus 2 after previous negative results might be attributed to false-negative laboratory results and prolonged viral shedding, rather than to re-infection. These findings are encouraging and should be validated in a larger cohort.

Polymerase chain reaction (PCR) testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is necessary for confirming a diagnosis of Coronavirus disease 2019 (COVID-19). Here we present a COVID-19 case of an elderly woman whose SARS-CoV-2 PCR tests showed false negative repeatedly by evaluating with different sampling sites and procedures. Nasopharyngeal swabs, suctioned sputum, and tongue swabs were collected for SARS-CoV-2-PCR. As for tongue swabs, we compared between two different sample conditions; one obtained with dry condition and the other obtained with moistened condition inside the oral cavity. SARS-CoV-2-PCR showed positive for an extended period with suctioned sputum samples compared with nasopharyngeal swabs and tongue swabs. No SARS-CoV-2 from a nasopharyngeal swab sample obtained on day 46 after symptoms onset was isolated despite high viral load (183740.5 copies/5μL). An adequate production of neutralizing antibody in a serum sample on day 46 was also confirmed. The number of RNA copies of the tongue swab samples was higher with moistened condition than with dry condition. The present case suggests that the difference of sampling site or sample condition can affect PCR results. High loads viral RNA detection does not always correlate with infectivity., (Copyright © 2021. Published by Elsevier Ltd.)

The coronavirus disease 2019 (COVID-19) outbreak started in December 2019 and rapidly spread around the globe as a major health threat. Several reports on re-positive cases subsequent to discharge from hospitals caught our attention. We aimed to highlight RT-qPCR positivity re-detection after discharge from isolation, with special consideration of the possible reasons behind it. We found that re-positive RT-qPCR assays for severe acute respiratory syndrome coronavirus 2 after previous negative results might be attributed to false-negative laboratory results and prolonged viral shedding, rather than to re-infection. These findings are encouraging and should be validated in a larger cohort., (© 2020 The Authors.)

We here report a case of coronavirus disease-19 (COVID-19) in Japan in which the initial throat swab polymerase chain reaction result was negative The possibility of false-negative results in the early phase of disease suggest reconsideration of the feasibility of a community or national infection control framework to prevent transmission. We recommend establishing an alternative feasible system, such as self-isolation by contact history in non-endemic community and by symptoms in endemic community, not relying on the PCR examination, to minimize this ongoing COVID-19 outbreak. Further rapid accumulation of knowledge including incubation period, clinical course and types of transmission is warranted to control this outbreak., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Annual antigen testing is a mainstay for diagnosing infection with Dirofiliaria immitis in dogs; yet, it has been documented that some heartworm-infected dogs and cats test false-negative for antigen due to the presence of antigen-antibody complexes. Several studies have reported the use of heat as a reliable means of immune-complex dissociation (ICD) in recent years; however, the data regarding the use of acid as a reliable method of ICD for D. immitis detection are limited. The objective of this study was to compare an acid-based form of ICD to the more established and evaluated method of heat-based ICD in experimentally infected and non-infected dogs. Plasma from class A dogs experimentally infected ∼4 months prior with D. immitis (infected; n = 24) and dogs reared indoors with no history of exposure to mosquitoes (non-infected; n = 75) were evaluated for presence of D. immitis antigen (DiroCHEK® Heartworm antigen test kit). Each sample was divided into three aliquots for testing: [1] Control plasma (no acid- or heat-treatment), [2] acid-treated plasma (trichloroacetic acid (TCA), incubation, centrifugation for 5 min at 16,000 X g, buffer), and [3] heat-treated plasma (104 °C followed by centrifugation at 16,000 X g). Treatments for each aliquot were performed and tested in triplicate; results were determined both visually (color change) and by spectrophotometric analysis (optical density [OD] value). Of the 24 infected dogs, 0/24 tested positive for antigen in the absence of acid- or heat-treatment. Those same plasma samples following processing by either acid- or heat-treatment yielded 18/24 (75.0%) and 19/24 (79.2%) antigen-positive results, respectively. Of the 75 plasma samples from non-infected dogs, neither acid- nor heat-treatment of plasma caused any false-positive color changes or spectrophotometric values. These results indicate that acid as a means of ICD reliably allowed for the detection of D. immitis antigen in infected plasma while not inducing false-positive results in non-infected plasma samples., (Copyright © 2020 Elsevier B.V. All rights reserved.)

There has been an increasing interest in accurate prediction of human pharmacokinetics of drug candidates to reduce cost and increase productivity during research and development. Modeling efforts have primarily focused on predicting drug absorption after oral administration because it is the most desired route for small molecule drug delivery. Despite significant progress in the field, the fraction of dose absorbed (Fa) is still considered to be a challenging parameter to predict. In recent years, compartment and transit models have become increasingly popular because of their effectiveness. A multicompartment plug flow-based model in which the stomach is assigned as a single compartment has been built. However, this model was found to be less accurate in estimating stomach solubilization of basic drugs under certain conditions and leads to false-negative results. Therefore, a modified multicompartment approach was developed by dividing the stomach into 4 compartments allowing the model to better mimic the physiological conditions. This approach was found to be more precise in estimating Fa for basic drugs compared to the previous approach. Based on this finding, it is believed the aforementioned approach should also be applicable to all compartmental-based models when similar issues are encountered., (Copyright © 2020 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

Human immunodeficiency virus (HIV)/AIDS is a major public health problem worldwide. Here we report the case of a patient with positive HIV-1 serology and negative result of The COBAS AmpliPrep/COBAS Taqman HIV-1 Test version 2.0 (CAP/CTM v2.0) assay (Roche Diagnostics) for HIV-1 RNA quantification in the absence of antiretroviral therapy (ARV). Considering the HIV contact history and repeating serological confirmatory test is essential before considering a low or even undetectable HIV-1 viral load to be a false-negative result., (Copyright © 2018. Published by Elsevier Ltd.)

The detection of hotspot mutations in key cancer genes is now an essential part of the diagnostic work-up in molecular pathology. Nearly all assays for mutation detection involve an amplification step. A second single nucleotide variant (SNV) on the same allele adjacent to a mutational hotspot can interfere with primer binding, leading to unnoticed allele-specific amplification of the wild type allele and thereby false-negative mutation testing. We present two diagnostic cases with false negative sequence results for JAK2 and SRSF2. In both cases mutations would have escaped detection if only one strand of DNA had been analysed. Because many commercially available diagnostic kits rely on the analysis of only one DNA strand they are prone to fail in cases like these. Detailed protocols and quality control measures to prevent corresponding pitfalls are presented., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Cannabis sativa subspecies, known as industrial hemp (C. sativa sativa) and marijuana (C. sativa indica) show no evident morphological distinctions, but they contain different levels of psychoactive Δ-9-tetrahidrocanabinol (THC), with considerably higher concentration in marijuana than in hemp. C. sativa subspecies differ in sequence of tetrahydrocannabinolic acid (THCA) synthase gene, responsible for THC production, and only one active copy of the gene, distinctive for marijuana, is capable of producing THC in concentration more then 0,3% in dried plants, usually punishable by the law. Twenty different samples of marijuana that contain THC in concentration more then 0,3% and three varieties of industrial hemp were analyzed for presence of an active copy of THCA synthase gene using in-house developed restriction fragment length polymorphism (RFLP) method All twenty samples of marijuana were positive for the active copy of THCA synthase gene, 16 of them heterozygous. All three varieties of industrial hemp were homozygous for inactive copy. An algorithm for the fast and accurate forensic analysis of samples suspected to be marijuana was constructed, answering the question if an analyzed sample is capable of producing THC in concentrations higher than 0.3%., (Copyright © 2017 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.)

Objectives: This study sought to investigate the clinical and imaging characteristics associated with diagnostic inaccuracy of computed tomography angiography (CTA) for detecting obstructive coronary artery disease (CAD) defined by quantitative coronary angiography (QCA)., Background: Although diagnostic performance metrics of CTA have been reported, there are sparse data on predictors of diagnostic inaccuracy by CTA., Methods: The clinical characteristics of 291 patients (mean age: 59 ± 10 years; female: 25.8%) enrolled in the multicenter CorE-64 (Coronary Artery Evaluation Using 64-Row Multi-detector Computed Tomography Angiography) study were examined. Pre-defined CTA segment-level characteristics of all true-positive (N = 237), false-positive (N = 115), false-negative (FN) (N = 159), and a random subset of true-negative segments (N = 511) for ≥50% stenosis with QCA as the reference standard were blindly abstracted in a central core laboratory. Factors independently associated with corresponding levels of CTA diagnostic inaccuracies on a patient level and coronary artery segment level were determined using multivariable logistic regression models and generalized estimating equations, respectively., Results: An Agatston calcium score of ≥1 per patient (odds ratio [OR]: 5.2; 95% confidence interval [CI]: 1.1 to 24.6) and the presence of within-segment calcification (OR: 10.2; 95% CI: 5.2 to 19.8) predicted false-positive diagnoses. Conversely, absence of within-segment calcification was an independent predictor of an FN diagnosis (OR: 2.0; 95% CI: 1.2 to 3.5). Prior percutaneous revascularization was independently associated with patient-level misdiagnosis of obstructive CAD (OR: 4.2; 95% CI: 1.6 to 11.2). Specific segment characteristics on CTA, notably segment tortuosity (OR: 3.5; 95% CI: 2.4 to 5.1), smaller luminal caliber (OR: 0.48; 95% CI: 0.36 to 0.63 per 1-mm increment), and juxta-arterial vein conspicuity (OR: 2.1; 95% CI: 1.4 to 3.2), were independently associated with segment-level misdiagnoses. Attaining greater intraluminal contrast enhancement independently lowered the risk of an FN diagnosis (OR: 0.96; 95% CI: 0.94 to 0.99 per 10-Hounsfield unit increment)., Conclusions: We identified clinical and readily discernible imaging characteristics on CTA predicting inaccurate CTA diagnosis of obstructive CAD defined by QCA. Knowledge and appropriate considerations of these features may improve the diagnostic accuracy in clinical CTA interpretation. (Diagnostic Accuracy of Multi-Detector Spiral Computed Tomography Angiography Using 64 Detectors [CORE-64]; NCT00738218)., (Copyright © 2013 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.)