Sensitive Determination of Infectious Titer of Recombinant Adeno-Associated Viruses (rAAVs) Using TCID50 End-Point Dilution and Quantitative Polymerase Chain Reaction (qPCR)

2020 ◽  
Vol 2020 (5) ◽  
pp. pdb.prot095653
Author(s):  
Martin Lock ◽  
James Wilson ◽  
Miguel Sena-Esteves ◽  
Guangping Gao
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
End Point ◽  
Infectious Titer ◽  
Sensitive Determination ◽  
Polymerase Chain

Sensitive Determination of the Rh D Genotype in Mixed Samples Using Fluorescence-Based Polymerase Chain Reaction

Vox Sanguinis ◽  
1997 ◽  
Vol 72 (3) ◽  
pp. 177-181 ◽  
Author(s):  
T. Tonn ◽  
D. Westrup ◽  
C. Seidl ◽  
C.M. Kirchmair ◽  
E. Seifried
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Sensitive Determination ◽  
Polymerase Chain ◽  
Mixed Samples

scholarly journals Technical Validation of a Reverse-Transcription Quantitative Polymerase Chain Reaction In Vitro Diagnostic Test for the Determination of MiR-31-3p Expression Levels in Formalin-Fixed Paraffin-Embedded Metastatic Colorectal Cancer Tumor Specimens

2018 ◽  
Vol 13 ◽  
pp. 117727191876335 ◽  
Author(s):  
Lucas Ramon ◽  
Catherine David ◽  
Karine Fontaine ◽  
Elodie Lallet ◽  
Charles Marcaillou ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Formalin Fixed

MiR-31-3p expression has been shown to be a predictive biomarker for response to anti-epithelial growth factor receptor therapy in patients with RAS wild-type metastatic colorectal cancer (mCRC). To aid in the quantification of miR-31-3p expression in formalin-fixed paraffin-embedded (FFPE) primary tumor samples from patients with mCRC, a reverse-transcription quantitative polymerase chain reaction (RT-qPCR) assay was developed and validated. Assay development included the identification of a microRNA reference standard and the determination of an appropriate relative quantification cutoff for differentiating low versus high miR-31-3p expression. Sample specimens for the validation studies included both FFPE slides and shavings. Polymerase chain reaction (PCR) efficiency and linearity, analytical sensitivity and specificity, assay robustness, reproducibility, and accuracy were demonstrated across a number of test conditions and differing quantitative PCR platforms. The data from this study provide evidence as to the feasibility of quantifying the expression of miR-31-3p from FFPE tumor tissue using a standardized RT-qPCR assay.

Rapid and accurate determination of the potency of varicella vaccine by quantitative polymerase chain reaction

Vaccine ◽  
2011 ◽  
Vol 29 (47) ◽  
pp. 8490-8495 ◽  
Author(s):  
Marsha S. Russell ◽  
Changgui Li ◽  
Louise Larocque ◽  
Junzhi Wang ◽  
Aaron Farnsworth ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Accurate Determination ◽  
Varicella Vaccine ◽  
Chain Reaction ◽  
Polymerase Chain

Sensitive Determination of the Rh D Genotype in Mixed Samples Using Fluorescence-Based Polymerase Chain Reaction

Vox Sanguinis ◽  
1997 ◽  
Vol 72 (3) ◽  
pp. 177-181 ◽  
Author(s):  
T. Tonn ◽  
D. Westrup ◽  
C. Seidl ◽  
C. M. Kirchmaier ◽  
E. Seifried
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Sensitive Determination ◽  
Polymerase Chain ◽  
Mixed Samples

Determination of carrier status in Duchenne and Becker muscular dystrophies by quantitative polymerase chain reaction and allele-specific oligonucleotides

1990 ◽  
Vol 36 (12) ◽  
pp. 2113-2117 ◽  
Author(s):  
T W Prior ◽  
A C Papp ◽  
P J Snyder ◽  
W E Highsmith ◽  
K J Friedman ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Muscular Dystrophy ◽  
Gene Dosage ◽  
Quantitative Polymerase Chain Reaction ◽  
Becker Muscular Dystrophy ◽  
Carrier Status ◽  
Chain Reaction ◽  
Allele Specific ◽  
Polymerase Chain

Abstract Detection of carriers of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), in the deletion cases, involves calculating gene dosage from Southern blots. We show that the analysis of dosage can also be made from the polymerase chain reaction (PCR) with use of allele-specific oligonucleotides (ASOs). The deletion-prone exons are amplified, transferred to a membrane, and hybridized with ASOs complementary to the exons; the autoradiographic bands are then quantified with a densitometer. After determining the quantitative conditions of the amplification reaction, we were able to identify deletions in a DMD/BMD carrier female. The determination of carrier status via PCR removes several of the technical limitations of Southern analysis and is also cost- and labor-effective.

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