scholarly journals Extraction of Viral Nucleic Acids with Carbon Nanotubes Increases SARS-CoV-2 Quantitative Reverse Transcription Polymerase Chain Reaction Detection Sensitivity

ACS Nano ◽  
2021 ◽  
Author(s):  
Sanghwa Jeong ◽  
Eduardo González-Grandío ◽  
Nicole Navarro ◽  
Rebecca L. Pinals ◽  
Francis Ledesma ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Polymerase Chain Reaction ◽  
Nucleic Acids ◽  
Reverse Transcription ◽  
Detection Sensitivity ◽  
Polymerase Chain Reaction Detection ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Specific Detection of Potato Virus A in Dormant Tubers by Reverse-Transcription Polymerase Chain Reaction

Plant Disease ◽  
1998 ◽  
Vol 82 (2) ◽  
pp. 230-234 ◽  
Author(s):  
Rudra P. Singh ◽  
Mathuresh Singh
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acids ◽  
Potato Virus ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Specific Detection ◽  
Chain Reaction ◽  
Breeding Lines ◽  
Potato Virus A ◽  
Polymerase Chain

A reverse-transcription polymerase chain reaction (RT-PCR) protocol was developed for the detection of potato virus A (PVA) in dormant tubers. A 255-bp amplified product was produced using a primer pair from the P1 gene of the PVA genome. The 255-bp product was detected in nucleic acids from leaves, tubers, and purified virions and was specific to PVA as determined by Southern blot tests and detection by a PVA-specific probe. When presented with seven potato virus/strain nucleic acids and a viroid, singly and in mixed infections, the primer pair did not amplify any products. Its specificity to PVA was further demonstrated by RT-PCR detection of PVA from the known mixtures of PVA and potato virus Y samples. PVA was detected in foliage nucleic acids at a dilution of 1:1024–1:4096 and tuber nucleic acids at 1:256–1:1024. It was uniformly present in various parts of the potato tuber. PVA was detected in composite tuber samples containing a ratio of infected to healthy sap of 1:29 and was readily detected in tubers of several cultivars or breeding lines, in dormant as well as in sprouting tubers stored at 20–25°C for 4 months.

scholarly journals A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay With Enhanced Capacity to Detect Vesicular Stomatitis Viral Lineages of Central American Origin

2021 ◽  
Vol 8 ◽  
Author(s):  
Kate Hole ◽  
Charles Nfon ◽  
Luis L. Rodriguez ◽  
Lauro Velazquez-Salinas
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sensitivity And Specificity ◽  
Reverse Transcription ◽  
Polymerase Chain Reaction Assay ◽  
Detection Sensitivity ◽  
Analytical Sensitivity ◽  
Chain Reaction ◽  
Vesicular Stomatitis ◽  
Polymerase Chain

Vesicular stomatitis virus (VSV) causes a disease in susceptible livestock that is clinically indistinguishable from foot-and-mouth disease. Rapid testing is therefore critical to identify VSV and rule out FMD. We previously developed and validated a multiplex real-time reverse transcription polymerase chain reaction assay (mRRT-PCR) for detection of both VS New Jersey virus (VSNJV) and VS Indiana virus (VSIV). However, it was subsequently apparent that this assay failed to detect some VSNJV isolates in Mexico, especially in genetic group II, lineage 2.1. In order to enhance the sensitivity of the mRRT-PCR for VSNJV, parts of the assay were redesigned and revalidated using new and improved PCR chemistries. The redesign markedly improved the assay by increasing the VSNJV detection sensitivity of lineage 2.1 and thereby allowing detection of all VSNJV clades. The new assay showed an increased capability to detect VSNJV. Specifically, the new mRRT-PCR detected VSNJV in 100% (87/87) of samples from Mexico in 2006-2007 compared to 74% for the previous mRRT-PCR. Furthermore, the analytical sensitivity of the new mRRT-PCR was enhanced for VSNJV. Importantly, the modified assay had the same sensitivity and specificity for VSIV as the previously published assay. Our results highlight the challenges the large genetic variability of VSV pose for virus detection by mRRT-PCR and show the importance of frequent re-evaluation and validation of diagnostic assays for VSV to ensure high sensitivity and specificity.

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