@article{PAR00006593,
  title = {{RNA} and {DNA} bacteriophages as molecular diagnosis controls in clinical virology : a comprehensive study of more than 45,000 routine {PCR} tests},
  author = {{N}inove, {L}. and {N}ougair{\`e}de, {A}. and {G}azin, {C}. and {T}hirion, {L}. and {D}elogu, {I}. and {Z}andotti, {C}. and {C}harrel, {R}. {N}. and de {L}amballerie, {X}avier},
  editor = {},
  language = {{ENG}},
  abstract = {{R}eal-time {PCR} techniques are now commonly used for the detection of viral genomes in various human specimens and require for validation both external and internal controls ({EC}s and {IC}s). {I}n particular, {IC}s added to clinical samples are necessary to monitor the extraction, reverse transcription, and amplification steps in order to detect false-negative results resulting from {PCR}-inhibition or errors in the technical procedure. {H}ere, we performed a large scale evaluation of the use of bacteriophages as {IC}s in routine molecular diagnosis. {T}his allowed to propose simple standardized procedures (i) to design specific {EC}s for both {DNA} and {RNA} viruses and (ii) to use {T}4 ({DNA}) or {MS}2 ({RNA}) phages as {IC}s in routine diagnosis. {V}arious technical formats for using phages as {IC}s were optimised and validated. {S}ubsequently, {T}4 and {MS}2 {IC}s were evaluated in routine real-time {PCR} or {RT}-{PCR} virological diagnostic tests, using a series of 8,950 clinical samples (representing 36 distinct specimen types) sent to our laboratory for the detection of a variety of {DNA} and {RNA} viruses. {T}he frequency of inefficient detection of {IC}s was analyzed according to the nature of the sample. {I}nhibitors of enzymatic reactions were detected at high frequency in specific sample types such as heparinized blood and bone marrow (>70%), broncho-alveolar liquid (41%) and stools (36%). {T}he use of {T}4 and {MS}2 phages as {IC}s proved to be cost-effective, flexible and adaptable to various technical procedures of real-time {PCR} detection in virology. {I}t represents a valuable strategy for enhancing the quality of routine molecular diagnosis in laboratories that use in-house designed diagnostic systems, which can conveniently be associated to the use of specific synthetic {EC}s. {T}he high rate of inhibitors observed in a variety of specimen types should stimulate the elaboration of improved technical protocols for the extraction and amplification of nucleic acids.},
  keywords = {},
  booktitle = {},
  journal = {{P}los {O}ne},
  volume = {6},
  numero = {2},
  pages = {e16142},
  ISSN = {1932-6203},
  year = {2011},
  DOI = {10.1371/journal.pone.0016142},
  URL = {https://www.documentation.ird.fr/hor/{PAR}00006593},
}