scholarly journals The Effect of Genomic DNA Contamination on the Detection of Circulating Long Non-Coding RNAs: The Paradigm of MALAT1

Diagnostics ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1160
Author(s):  
Athina N. Markou ◽  
Stavroula Smilkou ◽  
Emilia Tsaroucha ◽  
Evi Lianidou

The presence of contaminating gDNA in RNA preparations is a frequent cause of false positives in RT-PCR-based analysis. However, in some cases, this cannot be avoided, especially when there are no exons–intron junctions in the lncRNA sequences. Due to the lack of exons in few of long noncoding RNAs (lncRNAs) and the lack of DNAse treatment step in most studies reported so far, serious questions are raised about the specificity of lncRNA detection and the potential of reporting false-positive results. We hypothesized that minute amounts of gDNA usually co-extracted with RNA could give false-positive signals since primers would specifically bind to gDNA due to the lack of junction. In the current study, we evaluated the effect of gDNA and other forms of DNA like extrachromosomal circular DNAs (eccDNAs) contamination and the importance of including a DNAse treatment step on lncRNAsexpression.As a model, we have chosen as one of the most widely studied lncRNAs in cancer namely MALAT1, which lacks exons. When we tested this hypothesis in plasma and primary tissue samples from NSCLC patients, our findings clearly indicated that results on MALAT1 expression are highly affected by the presence of DNA contamination and that the DNAse treatment step is absolutely necessary to avoid false positive results.


1974 ◽  
Vol 31 (02) ◽  
pp. 273-278
Author(s):  
Kenneth K Wu ◽  
John C Hoak ◽  
Robert W Barnes ◽  
Stuart L Frankel

SummaryIn order to evaluate its daily variability and reliability, impedance phlebography was performed daily or on alternate days on 61 patients with deep vein thrombosis, of whom 47 also had 125I-fibrinogen uptake tests and 22 had radiographic venography. The results showed that impedance phlebography was highly variable and poorly reliable. False positive results were noted in 8 limbs (18%) and false negative results in 3 limbs (7%). Despite its being simple, rapid and noninvasive, its clinical usefulness is doubtful when performed according to the original method.



1995 ◽  
Vol 31 (5-6) ◽  
pp. 403-406 ◽  
Author(s):  
E. Frahm ◽  
U. Obst

Two recently developed Legionella detection tests, a microbiological-immunological method based on monoclonal antibodies (carried out as a colony-blot assay) and a commercial gene-probe testkit (the EnvironAmp Legionella Kit), are compared with the standard method. The colony-blot assay is faster than the conventional method; the gene-probe test is much faster still and is the most sensitive, but in consequence is at greater risk of false-positive results.



Author(s):  
Kazuhiro Itoh ◽  
Toru Kawamitsu ◽  
Yoko Osaka ◽  
Kazuyo Sato ◽  
Yusuke Suzuki ◽  
...  


Author(s):  
Takahiro Takazono ◽  
Tomomi Saijo ◽  
Nobuyuki Ashizawa ◽  
Kazuhiro Oshima ◽  
Keitaro Nishimura ◽  
...  


2012 ◽  
Vol 7 (6) ◽  
pp. 562-571 ◽  
Author(s):  
Roger Giner-Sorolla

The current crisis in psychological research involves issues of fraud, replication, publication bias, and false positive results. I argue that this crisis follows the failure of widely adopted solutions to psychology’s similar crisis of the 1970s. The untouched root cause is an information-economic one: Too many studies divided by too few publication outlets equals a bottleneck. Articles cannot pass through just by showing theoretical meaning and methodological rigor; their results must appear to support the hypothesis perfectly. Consequently, psychologists must master the art of presenting perfect-looking results just to survive in the profession. This favors aesthetic criteria of presentation in a way that harms science’s search for truth. Shallow standards of statistical perfection distort analyses and undermine the accuracy of cumulative data; narrative expectations encourage dishonesty about the relationship between results and hypotheses; criteria of novelty suppress replication attempts. Concerns about truth in research are emerging in other sciences and may eventually descend on our heads in the form of difficult and insensitive regulations. I suggest a more palatable solution: to open the bottleneck, putting structures in place to reward broader forms of information sharing beyond the exquisite art of present-day journal publication.



Author(s):  
Timo Huber ◽  
Philipp Steininger ◽  
Pascal Irrgang ◽  
Klaus Korn ◽  
Matthias Tenbusch ◽  
...  

AbstractSARS-CoV-2 antibody assays are used for epidemiological studies and for the assessment of vaccine responses in highly vulnerable patients. So far, data on cross-reactivity of SARS-CoV-2 antibody assays is limited. Here, we compared four enzyme-linked immunosorbent assays (ELISAs; Vircell SARS-CoV-2 IgM/IgA and IgG, Euroimmun SARS-CoV-2 IgA and IgG) for detection of anti-SARS-CoV-2 antibodies in 207 patients with COVID-19, 178 patients with serological evidence of different bacterial infections, 107 patients with confirmed viral respiratory disease, and 80 controls from the pre-COVID-19 era. In COVID-19 patients, the assays showed highest sensitivity in week 3 (Vircell-IgM/A and Euroimmun-IgA: 78.9% each) and after week 7 (Vircell-IgG: 97.9%; Euroimmun-IgG: 92.1%). The antibody indices were higher in patients with fatal disease. In general, IgM/IgA assays had only limited or no benefit over IgG assays. In patients with non-SARS-CoV-2 respiratory infections, IgG assays were more specific than IgM/IgA assays, and bacterial infections were associated with more false-positive results than viral infections. The specificities in bacterial and viral infections were 68.0 and 81.3% (Vircell-IgM/IgA), 84.8 and 96.3% (Euroimmun-IgA), 97.8 and 86.0% (Vircell-IgG), and 97.8 and 99.1% (Euroimmun-IgG), respectively. Sera from patients positive for antibodies against Mycoplasma pneumoniae, Chlamydia psittaci, and Legionella pneumophila yielded particularly high rates of unspecific false-positive results in the IgM/IgA assays, which was revealed by applying a highly specific flow-cytometric assay using HEK 293 T cells expressing the SARS-CoV-2 spike protein. Positive results obtained with anti-SARS-CoV-2 IgM/IgA ELISAs require careful interpretation, especially if there is evidence for prior bacterial respiratory infections.



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