Reliable detection and quantitation of viral nucleic acids in oral fluid: Liquid phase-based sample collection in conjunction with automated and standardized molecular assays

2008 ◽  
Vol 80 (9) ◽  
pp. 1684-1688 ◽  
Author(s):  
Reinhard B. Raggam ◽  
Jasmin Wagner ◽  
Birgit D. A. Michelin ◽  
Csilla Putz-Bankuti ◽  
Andreas Lackner ◽  
...  
Keyword(s):  
Liquid Phase ◽  
Nucleic Acids ◽  
Oral Fluid ◽  
Sample Collection ◽  
Molecular Assays ◽  
Reliable Detection

scholarly journals Commercial Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Molecular Assays: Superior Analytical Sensitivity of cobas SARS-CoV-2 Relative to NxTAG CoV Extended Panel and ID NOW COVID-19 Test

2020 ◽  
Vol 144 (11) ◽  
pp. 1303-1310 ◽  
Author(s):  
Run Jin ◽  
Matthew A. Pettengill ◽  
Nicole L. Hartnett ◽  
Herbert E. Auerbach ◽  
Stephen C. Peiper ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Nucleic Acid Amplification ◽  
Analytical Sensitivity ◽  
E Gene ◽  
Molecular Assays ◽  
Performance Variation ◽  
Wide Range ◽  
Reliable Detection ◽  
Testing Algorithms ◽  
Low Levels

Context.— We implemented multiple nucleic acid amplification test platforms because of the limited availability of test kits for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the early stages of the pandemic. Interpretation of results generated by different platforms and prioritization for testing algorithms required cross-comparison. Objective.— To compare the analytical sensitivity of 3 commercial SARS-CoV-2 molecular assays, selected samples were studied in parallel with Cobas SARS-CoV-2 test, NxTAG CoV Extended Panel, and ID NOW COVID-19 assays. Design.— A total of 8043 SARS-CoV-2 tests performed from March 22 to April 19, 2020, were included in this study. For all 1794 positive specimens detected by the cobas SARS-CoV-2 assay, the cycle threshold (Ct) values were manually tracked and plotted to demonstrate the distribution of sample viral levels. Additionally, 50 and 63 low-positive specimens (Ct values >32) as well as 50 and 61 consecutive positive specimens by the cobas assay were tested with NxTAG and ID NOW, respectively, to estimate their relative sensitivities. Results.— The Ct values of cobas SARS-CoV-2–positive samples were evenly distributed throughout ranges of 13.32 to 39.50 (mean, 25.06) and 13.60 to 42.49 (mean, 26.45) for ORF1 and E gene targets, respectively. NxTAG reliably detected only specimens with E gene Ct values lower than 33, and is estimated to detect 89.4% of positive specimens detected by cobas assay. ID NOW had performance variation independent of Ct value and is estimated to detect 83.5% of cobas positives. Conclusions.— Clinical specimens exhibit a wide range of viral burden, with a significant portion at low levels. Analytical sensitivity of testing platforms is critical for reliable detection of SARS-CoV-2 and uniform care to patients.

scholarly journals Molecular assays for the detection of prostate tumor derived nucleic acids in peripheral blood

2010 ◽  
Vol 9 (1) ◽  
pp. 174 ◽  
Author(s):  
Matthias Jost ◽  
John R Day ◽  
Ryan Slaughter ◽  
Theodore D Koreckij ◽  
Deanna Gonzales ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Peripheral Blood ◽  
Prostate Tumor ◽  
Molecular Assays

THC and CBD concentrations in blood, oral fluid and urine following a single and repeated administration of “light cannabis”

2020 ◽  
Vol 58 (5) ◽  
pp. 682-689 ◽  
Author(s):  
Roberta Pacifici ◽  
Simona Pichini ◽  
Manuela Pellegrini ◽  
Maria Concetta Rotolo ◽  
Raffaele Giorgetti ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Oral Fluid ◽  
Repeated Administration ◽  
Urine Samples ◽  
Gas Chromatography Mass Spectrometry ◽  
Sample Collection ◽  
Valuable Alternative ◽  
The Mean ◽  
Single And Repeated Administration

AbstractBackground“Light cannabis” is a product legally sold in Europe with Δ9-tetrahydrocannabinol (THC) concentration lower than 0.2% and variable cannabidiol (CBD) content. We studied THC and CBD excretion profiles in blood, oral fluid (OF) and urine after smoking one or four light cannabis cigarettes.MethodsBlood, OF and urine samples were obtained from six healthy light cannabis consumers after smoking one 1 g cigarette containing 0.16% THC and 5.8% CBD and from six others after smoking four 1 g cigarettes within 4 h. Sample collection began 0.5 and 4.5 h after smoking one or four cigarettes, respectively. Cannabinoid concentrations were quantified by gas chromatography-mass spectrometry (GC-MS).ResultsAt the first collection, the highest THC and CBD concentrations occurred in blood (THC 7.0–10.8 ng/mL; CBD 30.2–56.1 ng/mL) and OF (THC 5.1–15.5 ng/mL; CBD 14.2–28.1 ng/mL); similar results occurred 0.5 h after the last of four cigarettes in blood (THC 14.1–18.2 ng/mL, and CBD 25.6–45.4 ng/mL) and OF (THC 11.2–24.3 ng/mL; CBD 14.4–37.0 ng/mL). The mean OF to blood ratio ranged from 0.6 to 1.2 after one and 0.6 to 1.9 after four light cannabis cigarettes. THC/CBD ratios in blood and OF were never greater than 2. Urinary 11-nor-9-carboxy-THC concentrations peaked 8 h after one and four cigarettes.ConclusionsOF was a valuable alternative to blood in monitoring consumption of light cannabis. Blood and OF THC/CBD concentration ratios, never exceeded 2, possibly providing a useful biomarker to identify light cannabis vs illegal higher THC cannabis use, where THC/CBD ratios are generally greater than 10.

scholarly journals DETEKSI DINI DEMAM BERDARAH DENGUE DENGAN PEMERIKSAAN ANTIGEN NS1

2013 ◽  
Vol 3 (1) ◽  
Author(s):  
Mayer F. Wowor
Keyword(s):  
Nucleic Acids ◽  
Acute Phase ◽  
Vaccine Development ◽  
Early Stage ◽  
Serological Test ◽  
Clinical Care ◽  
Dengue Infection ◽  
Diagnostic Methods ◽  
Sample Collection ◽  
Ns1 Antigen

Abstract: Efficient and accurate diagnosis of dengue is of primary importance for clinical care, surveillance activities, outbreak control, pathogenesis, academic research, vaccine development, and clinical trials. Laboratory diagnostic methods for confirming dengue virus infection may involve detection of the viruses, viral nucleic acids, antigens and antibodies, or a combination of these techniques. After the onset of illness, the virus can be detected in serum, plasma, circulating blood cells, and other tissues for 4–5 days. During the early stage of the disease, virus isolation and the detection of viral nucleic acids or antigens, can be used to confirm the diagnosis of dengue infection. NS1 antigen appears as early as day 1 after the onset of fever, and declines to undetectable levels after day 5–6. At the end of the acute phase of infection, a serological test is the method of choice for diagnosis. A range of laboratory diagnostic methods has been developed to support patient management and disease control. The choice of diagnostic method depends on the purpose for which the test is done, available  laboratory facilities and technical expertise, costs, and the time of sample collection. Keywords: laboratory diagnostic methods, acute phase, NS1 antigen     Abstrak: Diagnosis dengue yang efisien dan akurat adalah hal yang paling penting  untuk proses perawatan di klinik, aktivitas surveilens, kontrol penularan penyakit, patogenesis, penelitian akademik, pengembangan vaksin dan percobaan-percobaan klinis. Metode diagnosis laboratorium untuk mengonfirmasi adanya infeksi virus dengue dapat meliputi deteksi virus dengue, asam nukleat virus, antigen dan antibodi, atau kombinasi dari teknik-teknik tersebut. Setelah serangan penyakit, virus dapat dideteksi dalam serum, plasma, sel-sel darah yang bersirkulasi, dan di jaringan lain dalam waktu 4-5 hari. Selama tahap awal dari penyakit, isolasi virus, asam nukleat virus, atau deteksi antigen dapat digunakan untuk diagnosis infeksi. Antigen NS1 muncul pada hari pertama setelah serangan demam dan menurun ke tingkat tidak terdeteksi setelah 5-6 hari. Pada akhir fase akut infeksi, serologi adalah metode pilihan untuk diagnosis. Metode diagnosis laboratorium telah berkembang untuk menunjang penanganan pasien dan kontrol penyakit. Pilihan untuk metode diagnosis bergantung pada tujuan tes dilakukan, fasilitas laboratorium dan tenaga ahli yang tersedia, biaya, dan saat sampel dikumpulkan. Kata kunci: metode diagnosis laboratorium, fase akut, antigen NS1

scholarly journals Plasmodium falciparum Allelic Diversity: A Comparison of DNA Extraction from Isolates Collected on Rapid Diagnostic Tests (Rdts) and Filter Paper

2021 ◽  
Author(s):  
Denise Patricia Mawili-Mboumba ◽  
Marie Louise Tshibola Mbuyi ◽  
Noe Patrick M’bondoukwe ◽  
Marielle Karine Bouyou-Akotet
Keyword(s):  
Plasmodium Falciparum ◽  
Nucleic Acids ◽  
Filter Paper ◽  
Diagnostic Tests ◽  
Allelic Diversity ◽  
Storage Conditions ◽  
Rapid Diagnostic Tests ◽  
Sample Collection ◽  
Filter Papers

Background: To perform molecular epidemiologic studies based on large cohorts, material such as RDTs or filter papers are useful for biological sample collection and extraction of RNA or DNA of good quality. Thus, we aimed to assess the quality of DNA extracted from malaria rapid diagnostic tests (RDTs) stored at various temperatures for the analysis of Plasmodium falciparum genetic diversity. Methods: Febrile patients benefitted from free malaria diagnosis using microscopy in a malaria sentinel site, at the Regional Hospital Estuaire-Melen, in Gabon, in 2015. P. falciparum isolates were collected onto one filter paper and 2 similar RDTs devices (Acon®) per patient. Nucleic acids were extracted with QiAmp Qiagen kit from paper and RDTs and the quality of the DNA was analyzed by msp1 gene amplification. Results: Msp1gene amplification was achieved in nucleic acids extracted from all filter papers and RDTs devices (n = 45, 100%). K1 alleles were detected in 93.3% (n = 42/45) of the samples and Mad20 alleles in 73.3% (n = 33/45). The number and the intensity of K1 and/or Mad20 fragments were comparable according to the sample collection material and the storage conditions (room temperature vs -20°C) of the samples. The size of the fragments indicating allelic diversity was comparable in 80% (n=36) of the samples. Conclusion: These data show that RDTs are a valuable source of DNA for malaria parasite genetic polymorphism analysis. Storage conditions of the devices did not influence the quality of DNA extracted from RDTs device, although some alleles may be missed.

Blood and Oral Fluid Cannabinoid Profiles of Frequent and Occasional Cannabis Smokers

2021 ◽  
Author(s):  
Melissa A Hoffman ◽  
Jacqueline A Hubbard ◽  
Philip M Sobolesky ◽  
Breland E Smith ◽  
Raymond T Suhandynata ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Safety Concern ◽  
Biological Fluid ◽  
Driving Under The Influence ◽  
Sample Collection ◽  
Detection Rates ◽  
Specimen Collection ◽  
Final Sample ◽  
Cannabis Consumption ◽  
Per Se

Abstract Increased prevalence of cannabis consumption and impaired driving are a growing public safety concern. Some states adopted per se driving laws, making it illegal to drive with more than a specified ∆9-tetrahydrocannabinol (THC) blood concentration of THC in a biological fluid (typically blood). Blood THC concentrations decrease significantly (~90%) with delays in specimen collection, suggesting use of alternative matrices, such as oral fluid (OF). We characterized 10 cannabinoids’ concentrations, including THC metabolites, in blood and OF from 191 frequent and occasional users by LC–MS-MS for up to 6 h after ad libitum smoking. Subjects self-titrated when smoking placebo, 5.9 or 13.4% THC cannabis. Higher maximum blood THC concentrations (Cmax) were observed in individuals who received the 5.9% THC versus the 13.4% THC plant material. In blood, the Cmax of multiple analytes, including THC and its metabolites, were increased in frequent compared to occasional users, whereas there were no significant differences in OF Cmax. Blood THC remained detectable (≥5 ng/mL) at the final sample collection for 14% of individuals who smoked either the 5.9% or 13.4% THC cigarette, whereas 54% had detectable THC in OF when applying the same cutoff. Occasional and frequent cannabis users’ profiles were compared, THC was detectable for significantly longer in blood and OF from frequent users. Detection rates between frequent and occasional users at multiple per se cutoffs showed larger differences in blood versus OF. Understanding cannabinoid profiles of frequent and occasional users and the subsequent impact on detectability with current drug per se driving limits is important to support forensic interpretations and the development of scientifically supported driving under the influence of cannabis laws.

scholarly journals Antibody detection by agglutination–PCR (ADAP) enables early diagnosis of HIV infection by oral fluid analysis

2018 ◽  
Vol 115 (6) ◽  
pp. 1250-1255 ◽  
Author(s):  
Cheng-ting Tsai ◽  
Peter V. Robinson ◽  
Felipe de Jesus Cortez ◽  
Maria L. B. Elma ◽  
David Seftel ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Detection Threshold ◽  
Population Based ◽  
Antibody Detection ◽  
Analytical Sensitivity ◽  
Sample Collection ◽  
Clinical Sensitivity ◽  
Fluid Analysis ◽  
Hiv Antibodies ◽  
Anti Hiv

Oral fluid (OF) is a highly effective substrate for population-based HIV screening efforts, as it is noninfectious and significantly easier to collect than blood. However, anti-HIV antibodies are found at far lower concentrations in OF compared with blood, leading to poor sensitivity and a longer period of time from infection to detection threshold. Thus, despite its inherent advantages in sample collection, OF is not widely used for population screening. Here we report the development of an HIV OF assay based on Antibody Detection by Agglutination–PCR (ADAP) technology. This assay is 1,000–10,000 times more analytically sensitive than clinical enzyme-linked immunoassays (EIAs), displaying both 100% clinical sensitivity and 100% specificity for detecting HIV antibodies within OF samples. We show that the enhanced analytical sensitivity enables this assay to correctly identify HIV-infected individuals otherwise missed by current OF assays. We envision that the attributes of this improved HIV OF assay can increase testing rates of at-risk individuals while enabling diagnosis and treatment at an earlier time point.

scholarly journals CRISPR technology incorporating amplification strategies: molecular assays for nucleic acids, proteins, and small molecules

2021 ◽  
Author(s):  
Wei Feng ◽  
Ashley M. Newbigging ◽  
Jeffrey Tao ◽  
Yiren Cao ◽  
Hanyong Peng ◽  
...  
Keyword(s):  
Analytical Chemistry ◽  
Nucleic Acids ◽  
Small Molecules ◽  
Genome Engineering ◽  
Molecular Assays ◽  
Diagnostic Technology

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) protein systems revolutionize genome engineering and advance analytical chemistry and diagnostic technology.

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