@article{fdi:010062013,
  title = {{D}evelopment of generic {T}aqman {PCR} and {RT}-{PCR} assays for the detection of {DNA} and m{RNA} of beta-actin-encoding sequences in a wide range of animal species},
  author = {{P}iorkowski, {G}. and {B}aronti, {C}{\'e}cile and de {L}amballerie, {X}avier and de {F}abritus, {L}. and {B}ichaud, {L}. and {P}astorino, {B}. {A}. and {B}essaud, {M}.},
  editor = {},
  language = {{ENG}},
  abstract = {{A}s a member of the {E}uropean {V}irus {A}rchive ({EVA}) consortium, our laboratory is developing and maintaining a large collection of viruses. {T}his collection implies the use of a panel of cell lines originating from various animal species. {I}n order to make easier the handling of such a large panel of cell lines, wide spectrum real-time {PCR} and {RT}-{PCR} assays were developed to allow the detection and the quantification of {DNA} and m{RNA} of beta-actin, one of the most commonly used eukaryotic housekeeping genes. {B}y using two degenerated primers and a unique probe, these two assays were shown to detect nucleic acids of a panel of vertebrate and invertebrate cell lines commonly used in animal virology. {T}his panel included human, monkey, rodent, dog, pig, fish, batrachian, mosquito and tick cell lines. {A}dditionally, the two assays amplified successfully beta-actin-encoding sequences of sandflies. {S}ensitivity evaluation performed on synthetic {DNA} and {RNA} sequences showed that the two assays were very sensitive and suitable for accurate quantification. {T}he two assays constitute together a convenient method suitable for multiple purposes. {T}hey can be used for instance to estimate the amount of contaminating cellular genetic material prior to sequence-independent amplification of viral genomes achieved before high-throughput sequencing, to evaluate the efficiency of {DN}ase and/or {RN}ase treatments performed on cellular extract and to check nucleic acid extraction by using beta-actin-encoding sequences as endogenous control. {T}his assay will constitute a precious tool for virologists working with multiple cell lines or animal models.},
  keywords = {{ACTB} gene ; {A}ctin ; {H}ousekeeping gene ; {T}aqman assay ; {N}ext generation ; sequencing},
  booktitle = {},
  journal = {{J}ournal of {V}irological {M}ethods},
  volume = {202},
  numero = {},
  pages = {101--105},
  ISSN = {0166-0934},
  year = {2014},
  DOI = {10.1016/j.jviromet.2014.02.026},
  URL = {https://www.documentation.ird.fr/hor/fdi:010062013},
}