ArticleBMC molecular biology2008
Phi29 polymerase based random amplification of viral RNA as an alternative to random RT-PCR.
Article in BMC molecular biology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
46 citing papers in PubMed, 80 citations in OpenAlex.
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- Laboratory diagnosis of nonpolio enteroviruses: A review of the current literature.Biosafety and health · 2023Review
- Rapid whole genome sequencing methods for RNA viruses.Frontiers in microbiology · 2023Article
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- Extensive qPCR analysis reveals altered gene expression in middle ear mucosa from cholesteatoma patients.PloS one · 2020Article
- Complete Genome Sequences of Igbo-Ora and Babanki Alphavirus Strains Isolated in the Central African Republic in the 1960s and 1970s.Microbiology resource announcements · 2019Article
- Complete Genome Sequence of an Equine Arteritis Virus Strain Isolated from a Lipizzaner Stallion in 2015 in Serbia.Microbiology resource announcements · 2019Article
- Complete Genome Sequence of the Tataguine Virus, Isolated in the Central African Republic in 1972 from a Human with an Acute Febrile Syndrome.Microbiology resource announcements · 2019Article
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- Limited reverse transcriptase activity of phi29 DNA polymerase.Nucleic acids research · 2018Article
- Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals [corrected].NPJ vaccines · 2018Article
- Complete Genome Sequences of Two Chikungunya Viruses Isolated in the Central African Republic in the 1970s and 1980s.Genome announcements · 2017Article
- African Non-Human Primates Host Diverse Enteroviruses.PloS one · 2017Article
- Virome analysis of two sympatric bat species (Desmodus rotundus and Molossus molossus) in French Guiana.PloS one · 2017Article
- Improved assembly procedure of viral RNA genomes amplified with Phi29 polymerase from new generation sequencing data.Biological research · 2016Article
- Viral metagenomics and blood safety.Transfusion clinique et biologique : journal de la Societe francaise de transfusion sanguine · 2016Review
- Whole Genome Sequencing of Enterovirus species C Isolates by High-Throughput Sequencing: Development of Generic Primers.Frontiers in microbiology · 2016Article
- Sequence-independent amplification coupled with DNA microarray analysis for detection and genotyping of noroviruses.AMB Express · 2015Article
- Viral diversity in swine intestinal mucus used for the manufacture of heparin as analyzed by high-throughput sequencing.Biologicals : journal of the International Association of Biological Standardization · 2015Article
Corrections and comments
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Authors and funding
15 authors at 2 institutions in 2 countries.
Funding
Abstract
backgroundPhi29 polymerase based amplification methods provides amplified DNA with minimal changes in sequence and relative abundance for many biomedical applications. RNA virus detection using microarrays, however, can present a challenge because phi29 DNA polymerase cannot amplify RNA nor small cDNA fragments (<2000 bases) obtained by reverse transcription of certain viral RNA genomes. Therefore, ligation of cDNA fragments is necessary prior phi29 polymerase based amplification. We adapted the QuantiTect Whole Transcriptome Kit (Qiagen) to our purposes and designated the method as Whole Transcriptome Amplification (WTA).
resultsWTA successfully amplified cDNA from a panel of RNA viruses representing the diversity of ribovirus genome sizes. We amplified a range of genome copy numbers from 15 to 4 x 10(7) using WTA, which yielded quantities of amplified DNA as high as 1.2 microg/microl or 10(10) target copies. The amplification factor varied between 10(9) and 10(6). We also demonstrated that co-amplification occurred when viral RNA was mixed with bacterial DNA.
conclusionThis is the first report in the scientific literature showing that a modified WGA (WTA) approach can be successfully applied to viral genomic RNA of all sizes. Amplifying viral RNA by WTA provides considerably better sensitivity and accuracy of detection compared to random RT-PCR.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.