scholarly journals A multiplex reverse transcription PCR and automated electronic microarray assay for detection and differentiation of seven viruses affecting swine

2017 ◽  
Vol 65 (2) ◽  
pp. e272-e283 ◽  
Author(s):  
A. Erickson ◽  
M. Fisher ◽  
T. Furukawa-Stoffer ◽  
A. Ambagala ◽  
D. Hodko ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Microarray Assay ◽  
Reverse Transcription Pcr ◽  
Electronic Microarray

scholarly journals DNA Microarray to Monitor the Expression of MAGE-A Genes

2002 ◽  
Vol 48 (1) ◽  
pp. 25-34 ◽  
Author(s):  
Nathalie Zammatteo ◽  
Laurence Lockman ◽  
Francis Brasseur ◽  
Etienne De Plaen ◽  
Christophe Lurquin ◽  
...  
Keyword(s):  
Dna Microarray ◽  
Reverse Transcription ◽  
Pcr Amplification ◽  
Gene Probe ◽  
Microarray Assay ◽  
Reverse Transcription Pcr ◽  
Antitumor Immunotherapy ◽  
Consensus Primers ◽  
Pcr Microarray ◽  
Assay Detection

Abstract Background: The MAGE-A genes encode antigens that are of particular interest for antitumor immunotherapy because they are strictly tumor specific and are shared by many tumors. We developed a rapid method to identify the MAGE-A genes expressed in tumors. Methods: A low-density DNA microarray was designed to discriminate between the 12 MAGE-A cDNAs amplified by PCR with only one pair of consensus primers. The assay involved reverse transcription of total RNA with oligo(dT) primer, followed by PCR amplification and hybridization on a microarray. Amplification in the presence of Biotin-16-dUTP allowed subsequent detection of the amplicons on the microarray carrying 12 capture probes, each being specific for a MAGE-A gene. Probe–amplicon hybrids were detected by a streptavidin-based method. Results: PCR conditions were optimized for low detection limits and comparable amplification efficiencies among all MAGE-A nucleotide sequences. The microarray assay was validated with a panel of 32 samples, by comparison with well-established reverse transcription-PCR assays relying on amplification with primers specific for each gene. Virtually identical results were obtained with both methods, except for MAGE-A3 and MAGE-A5. Detection of MAGE-A5 was more sensitive with the microarray assay. Detection of MAGE-A3 was hampered by the presence of MAGE-A6, which is 98% identical: the MAGE-A3 capture probe cross-hybridized with MAGE-A6 amplicons because these sequences differed by only a single base. Conclusions: This post-PCR microarray assay could be useful to evaluate MAGE expression in tumors before therapeutic vaccinations with MAGE-A gene products.

scholarly journals Implementation of an in-house real-time reverse transcription-PCR assay to detect the emerging SARS-CoV-2 N501Y variants

2021 ◽  
pp. 104868
Author(s):  
Marielle BEDOTTO ◽  
Pierre-Edouard FOURNIER ◽  
Linda HOUHAMDI ◽  
Philippe COLSON ◽  
Didier RAOULT
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Pcr Assay ◽  
Reverse Transcription Pcr

scholarly journals Reverse Transcription-PCR Analysis of Bottled and Natural Mineral Waters for the Presence of Noroviruses

2003 ◽  
Vol 69 (11) ◽  
pp. 6541-6549 ◽  
Author(s):  
Gilbert Thierry Lamothe ◽  
Thierry Putallaz ◽  
Han Joosten ◽  
Joey D. Marugg
Keyword(s):  
Reverse Transcription ◽  
Membrane Filtration ◽  
Limit Of Detection ◽  
Mineral Waters ◽  
Pcr Analysis ◽  
Rna Sequences ◽  
Natural Mineral ◽  
Reverse Transcription Pcr ◽  
Viral Particles ◽  
Nonspecific Amplification

ABSTRACT A seminested reverse transcription-PCR method coupled to membrane filtration was optimized to investigate the presence of norovirus (NV) RNA sequences in bottled and natural mineral waters. The recovery of viral particles by filtration varied between 28 and 45%, while the limit of detection of the overall method ranged from 6 to 95 viral particles. The assay was broadly reactive, as shown by the successful detection of 27 different viral strains representing 12 common genotypes of NVs. A total of 718 bottled and natural mineral water samples were investigated, including 640 samples of finished, spring, and line products (mostly 1 to 1.5 liters), collected from 36 different water brands of various types and from diverse geographic origins over a 2-year period. In addition, 78 samples of larger volume (10 and 400 to 500 liters) and environmental swabs were investigated. From the 1,436 analyses that were performed for the detection of NVs belonging to genogroups I and II, 34 samples (2.44%) were presumptively positive by seminested RT-PCR. However, confirmation by DNA sequence analysis revealed that all presumptive positive results were either due to nonspecific amplification or to cross-contamination. In conclusion, these results do not provide any evidence for the presence of NV genome sequences in bottled waters.

Direct subtyping of HLA-B and -A genes by automated sequencing following reverse transcription-PCR

1996 ◽  
Vol 47 (1-2) ◽  
pp. 115
Author(s):  
AJ Knipper ◽  
J Enczmann ◽  
P Hakenberg ◽  
G Kögler ◽  
P Wernet
Keyword(s):  
Reverse Transcription ◽  
Reverse Transcription Pcr

scholarly journals Aromatic Hydroxylation of Indan by o-Xylene-Degrading Rhodococcus sp. Strain DK17

2009 ◽  
Vol 76 (1) ◽  
pp. 375-377 ◽  
Author(s):  
Dockyu Kim ◽  
Choong Hwan Lee ◽  
Jung Nam Choi ◽  
Ki Young Choi ◽  
Gerben J. Zylstra ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Ring Cleavage ◽  
Growth Substrate ◽  
Aromatic Moiety ◽  
Reverse Transcription Pcr ◽  
Aromatic Hydroxylation ◽  
Rhodococcus Sp

ABSTRACT The metabolically versatile Rhodococcus sp. strain DK17 utilizes indan as a growth substrate via the o-xylene pathway. Metabolite and reverse transcription-PCR analyses indicate that o-xylene dioxygenase hydroxylates indan at the 4,5 position of the aromatic moiety to form cis-indan-4,5-dihydrodiol, which is dehydrogenated to 4,5-indandiol by a dehydrogenase. 4,5-Indandiol undergoes ring cleavage by a meta-cleavage dioxygenase.

scholarly journals Increased Expression of Two Multidrug Transporter-Like Genes Is Associated with Ethidium Bromide and Ciprofloxacin Resistance in Mycoplasma hominis

2005 ◽  
Vol 49 (1) ◽  
pp. 421-424 ◽  
Author(s):  
S. Raherison ◽  
P. Gonzalez ◽  
H. Renaudin ◽  
A. Charron ◽  
C. Bébéar ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Ethidium Bromide ◽  
Reverse Transcription ◽  
Mycoplasma Hominis ◽  
Atp Binding ◽  
Multidrug Transporter ◽  
Reverse Transcription Pcr ◽  
Atp Binding Cassette Transporters ◽  
Resistant Strains ◽  
Ciprofloxacin Resistance

ABSTRACT Two genes, md1 and md2, coding for multidrug resistance ATP-binding cassette transporters were identified in Mycoplasma hominis PG21. Expression of these two genes, quantified by quantitative competitive reverse transcription-PCR, was significantly increased in ethidium bromide-resistant strains of M. hominis compared to that in M. hominis PG21.

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