A Highly Sensitive Quantitative PCR for the Detection of Bartonella bacilliformis by Targeting a Multiple-Copy DNA Segment

2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Hua-Wei Chen ◽  
Ai Mochida ◽  
Philip Ching ◽  
Chien-Chung Chao ◽  
Wei-Mei Ching
Keyword(s):  
Quantitative Pcr ◽  
Multiple Copy ◽  
Bartonella Bacilliformis ◽  
Highly Sensitive ◽  
Copy Dna

Development of a highly sensitive real-time immuno-PCR for the measurement of chloramphenicol in milk based on magnetic bead capturing

2014 ◽  
Vol 6 (23) ◽  
pp. 9340-9347 ◽  
Author(s):  
Xiaoqi Tao ◽  
Zhifei He ◽  
Xingyuan Cao ◽  
Jianzhong Shen ◽  
Hongjun Li
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Magnetic Bead ◽  
The Real ◽  
Highly Sensitive

The real-time immuno-quantitative PCR (RT-IPCR) schematic illustration of the determination of CAP in milk based on magnetic bead capturing.

scholarly journals Pan-Piscine Orthoreovirus (PRV) Detection Using Reverse Transcription Quantitative PCR

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1548
Author(s):  
Julie Zhao ◽  
Niccolò Vendramin ◽  
Argelia Cuenca ◽  
Mark Polinski ◽  
Laura M. Hawley ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Reverse Transcription ◽  
High Sensitivity ◽  
Test Results ◽  
Reverse Transcription Quantitative Pcr ◽  
Highly Sensitive ◽  
Surveillance Programs ◽  
America South ◽  
Universal Detection ◽  
Quantitative Pcr Assay

Piscine orthoreovirus (PRV) infects farmed and wild salmon and trout species in North America, South America, Europe, and East Asia. PRV groups into three distinct genotypes (PRV-1, PRV-2, and PRV-3) that can vary in distribution, host specificity, and/or disease potential. Detection of the virus is currently restricted to genotype specific assays such that surveillance programs require the use of three assays to ensure universal detection of PRV. Consequently, herein, we developed, optimized, and validated a real-time reverse transcription quantitative PCR assay (RT-qPCR) that can detect all known PRV genotypes with high sensitivity and specificity. Targeting a conserved region at the 5′ terminus of the M2 segment, the pan-PRV assay reliably detected all PRV genotypes with as few as five copies of RNA. The assay exclusively amplifies PRV and does not cross-react with other salmonid viruses or salmonid host genomes and can be performed as either a one- or two-step RT-qPCR. The assay is highly reproducible and robust, showing 100% agreement in test results from an inter-laboratory comparison between two laboratories in two countries. Overall, as the assay provides a single test to achieve highly sensitive pan-specific PRV detection, it is suitable for research, diagnostic, and surveillance purposes.

Quantitative PCR for detection and discrimination of the bloodborne pathogen Staphylococcus epidermidis in platelet preparations using divIVA and icaA as target genes

2007 ◽  
Vol 53 (11) ◽  
pp. 1222-1231 ◽  
Author(s):  
Cherie Cameron Mastronardi ◽  
Sandra Ramírez-Arcos
Keyword(s):  
Quantitative Pcr ◽  
Internal Control ◽  
Staphylococcus Epidermidis ◽  
Target Genes ◽  
White Blood Cells ◽  
Virulence Gene ◽  
Qpcr Assay ◽  
Platelet Storage ◽  
Highly Sensitive ◽  
Internal Sequence

Bacterial contamination of blood components is the major microbiological cause of transfusion-associated morbidity, with Staphylococcus epidermidis being the most frequently isolated organism from contaminated platelet preparations (PPs). We have recently shown that S. epidermidis forms biofilms during platelet storage, which might account for reported missed detection during routine screening. In this study, we developed a highly sensitive and specific multiplex quantitative PCR (QPCR) assay to detect S. epidermidis in PPs at levels of 102–103 cfu/mL. A specific primer pair and hydrolysis probe were designed to amplify an internal region of the cell division divIVA gene that is unique to S. epidermidis. In addition, an internal sequence of the virulence gene icaA, which is involved in the synthesis of the S. epidermidis biofilm matrix, was selected to allow for differentiation of potentially biofilm-forming S. epidermidis isolates. A conserved region of the 8 alleles of the HLA-DQα1 locus present in residual white blood cells in PPs was selected as an internal control for the assay. The specificity of this assay was confirmed, as other staphylococcal species that were tested with the optimized parameters were not detected. This QPCR assay could be adaptable for the detection of other bloodborne bacterial pathogens.

Purification and structural analysis of interferon

1982 ◽  
Vol 299 (1094) ◽  
pp. 39-50 ◽  
Keyword(s):  
Biochemical Characterization ◽  
Structural Information ◽  
Scale Up ◽  
Structural Data ◽  
Highly Sensitive ◽  
Complete Sequences ◽  
Degree Of Confidence ◽  
Copy Dna ◽  
High Degree ◽  
Cdna Fragments

Studies with crude or partly purified interferon have provided a significant amount of structural information. However, complete biochemical characterization required purification to homogeneity. Earlier work on fractionation has met with many difficulties because interferon was available only in minute quantities. A scale-up of production, adaptation of multi-step purification schemes, use of high-resolution separation techniques and highly sensitive analytical methods have yielded pure interferons and hence many structural data. Specific activities, amino-acid compositions, partial sequences and structural homologies of many interferons were determined. Finally, cloned copy DNA (cDNA) fragments derived from specific interferon mRNA, as well as isolated interferon genes, have been sequenced and the data were used to elucidate complete sequences of many interferons with a high degree of confidence.

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