scholarly journals Development of Reverse Transcriptase Polymerase Chain Reaction Primer Sets and Standard Positive Control Capable of Verifying False Positive for the Detection of Severe acute respiratory syndrome coronavirus 2

2021 ◽  
Vol 27 (4) ◽  
pp. 283-290
Author(s):  
Kyu Bong Cho
Keyword(s):  
Polymerase Chain Reaction ◽  
Severe Acute Respiratory Syndrome ◽  
Reverse Transcriptase ◽  
False Positive ◽  
Positive Control ◽  
Polymerase Chain Reaction Primer ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Primer Sets

scholarly journals Polymerase chain reaction primer sets for the detection of genetically diverse human sapoviruses

2020 ◽  
Vol 165 (10) ◽  
pp. 2335-2340
Author(s):  
Tomoichiro Oka ◽  
Seiji P. Yamamoto ◽  
Nobuhiro Iritani ◽  
Shigenori Sato ◽  
Chika Tatsumi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Primer ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Primer Sets

scholarly journals Experience With Pretravel Testing for SARS-CoV-2 at an Academic Medical Center

2021 ◽  
Vol 8 ◽  
pp. 237428952110102
Author(s):  
Katherine L. Imborek ◽  
Matthew D. Krasowski ◽  
Paul Natvig ◽  
Anna E. Merrill ◽  
Daniel J. Diekema ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Severe Acute Respiratory Syndrome ◽  
Reverse Transcriptase ◽  
Medical Center ◽  
Academic Medical Center ◽  
Immunoglobulin M ◽  
Chain Reaction ◽  
Academic Medical ◽  
Polymerase Chain ◽  
Positive Results

International travel has been a significant factor in the coronavirus disease 2019 pandemic. Many countries and airlines have implemented travel restrictions to limit the spread of the causative agent, severe acute respiratory syndrome coronavirus-2. A common requirement has been a negative reverse-transcriptase polymerase chain reaction performed by a clinical laboratory within 48 to 72 hours of departure. A more recent travel mandate for severe acute respiratory syndrome coronavirus-2 immunoglobulin M serology testing was instituted by the Chinese government on October 29, 2020. Pretravel testing for severe acute respiratory syndrome coronavirus-2 raises complications in terms of cost, turnaround time, and follow-up of positive results. In this report, we describe the experience of a multidisciplinary collaboration to develop a workflow for pretravel severe acute respiratory syndrome coronavirus-2 reverse-transcriptase polymerase chain reaction and immunoglobulin M serology testing at an academic medical center. The workflow primarily involved self-payment by patients and preferred retrieval of results by the patient through the electronic health record patient portal (Epic MyChart). A total of 556 unique patients underwent pretravel reverse-transcriptase polymerase chain reaction testing, with 13 (2.4%) having one or more positive results, a rate similar to that for reverse-transcriptase polymerase chain reaction testing performed for other protocol-driven asymptomatic screening (eg, inpatient admissions, preprocedural) at our medical center. For 5 of 13 reverse-transcriptase polymerase chain reaction positive samples, the traveler had clinical history, prior reverse-transcriptase polymerase chain reaction positive, and high cycle thresholds values on pretravel testing consistent with remote infection and minimal transmission risk. Severe acute respiratory syndrome coronavirus-2 immunoglobulin M was performed on only 24 patients but resulted in 2 likely false positives. Overall, our experience at an academic medical center shows the challenge with pretravel severe acute respiratory syndrome coronavirus-2 testing.

scholarly journals Enhanced Polymerase Chain Reaction Methods for Detecting and Quantifying Phytophthora infestans in Plants

2000 ◽  
Vol 90 (10) ◽  
pp. 1112-1119 ◽  
Author(s):  
Howard S. Judelson ◽  
Paul W. Tooley
Keyword(s):  
Polymerase Chain Reaction ◽  
Phytophthora Infestans ◽  
Positive Control ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Diagnostic Assays ◽  
Specificity And Sensitivity ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Diagnostic Applications

Sensitive and specific primer sets for polymerase chain reaction (PCR) for Phytophthora infestans, the oomycete that causes late blight of potato and tomato, were developed based on families of highly repeated DNA. The performance of these primers was compared to those developed previously for the internal transcribed spacer (ITS) of ribosomal DNA. The detection limit using the new primers is 10 fg of P. infestans DNA, or 0.02 nuclei. This is about 100 times more sensitive than ITS-directed primers. Nested polymerase chain reaction (PCR) allows the measurement of down to 0.1 fg of DNA using the new primers. To enhance the reliability of diagnostic assays, an internal positive control was developed using an amplification mimic. The mimic also served as a competitor for quantitative PCR, which was used to assess the growth of P. infestans in resistant and susceptible tomato. A key dimension of this study was that two laboratories independently checked the specificity and sensitivity of each primer set; differences were noted that should be considered when PCR is adopted for diagnostic applications in any system.

scholarly journals Rapid, Sensitive, and Specific Severe Acute Respiratory Syndrome Coronavirus 2 Detection: A Multicenter Comparison Between Standard Quantitative Reverse-Transcriptase Polymerase Chain Reaction and CRISPR-Based DETECTR

2020 ◽  
Author(s):  
Eelke Brandsma ◽  
Han J M P Verhagen ◽  
Thijs J W van de Laar ◽  
Eric C J Claas ◽  
Marion Cornelissen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Severe Acute Respiratory Syndrome ◽  
Reverse Transcriptase ◽  
N Gene ◽  
Analytical Sensitivity ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Ribonucleoprotein Complexes ◽  
Large Patient ◽  
Polymerase Chain

Abstract Background Recent advances in CRISPR-based diagnostics suggest that DETECTR, a combination of reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) and subsequent Cas12 bystander nuclease activation by amplicon-targeting ribonucleoprotein complexes, could be a faster and cheaper alternative to quantitative reverse-transcription polymerase chain reaction (qRT-PCR) without sacrificing sensitivity and/or specificity. Methods In this study, we compare DETECTR with qRT-PCR to diagnose coronavirus disease 2019 on 378 patient samples. Patient sample dilution assays suggest a higher analytical sensitivity of DETECTR compared with qRT-PCR; however, this was not confirmed in this large patient cohort, where we report 95% reproducibility between the 2 tests. Results These data showed that both techniques are equally sensitive in detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) providing additional value of DETECTR to the currently used qRT-PCR platforms. For DETECTR, different guide ribonucleic acids can be used simultaneously to obviate negative results due to mutations in N-gene. Lateral flow strips, suitable as a point-of-care test, showed a 100% correlation to the high-throughput DETECTR assay. More importantly, DETECTR was 100% specific for SARS-CoV-2 relative to other human coronaviruses. Conclusions Because there is no need for specialized equipment, DETECTR could be rapidly implemented as a complementary technically independent approach to qRT-PCR thereby increasing the testing capacity of medical microbiological laboratories and relieving the existent PCR platforms for routine non-SARS-CoV-2 diagnostic testing.

scholarly journals Investigation of Compatibility of SARS-CoV-2 RT-PCR Kits Containing Different Gene Targets During COVID-19 Pandemic

2020 ◽  
Author(s):  
Figen Sarıgul ◽  
Osman Doluca ◽  
Sıla Akhan ◽  
Murat Sayan
Keyword(s):  
Polymerase Chain Reaction ◽  
Severe Acute Respiratory Syndrome ◽  
Reverse Transcriptase ◽  
Accurate Diagnosis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Chi Square ◽  
Unknown Sample ◽  
Deming Regression ◽  
Polymerase Chain

ABSTRACTAimIn the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), reverse transcriptase-polymerase chain reaction (RT-PCR) technique is often used. We evaluated the compatibility of SARS-CoV-2 RT-PCR kits containing different gene targets during the pandemic.Materials & methodsSamples were tested by Bio-Speddy® (RdRp gene) and Diagnovital® (RdRp+E genes). The correlation between two assays were determined by Deming regression and chi-square heatmap analyses.ResultsDiagnovital PCR kit showed in a constricted range and conveniently exponential amplification curves than Bio-Speedy PCR kit. While the correlation increased when a secondary biomarker was added to the kit.ConclusionIn an unknown sample, using together different PCR kits that target different genes during the pandemic situation may provide a more accurate diagnosis of SARS-CoV-2.

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