Real-time reverse-transcription loop-mediated isothermal amplification for rapid detection of rift valley Fever virus.
The development and validation of a one-step, single-tube, real-time accelerated reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for the detection of the L RNA segment of Rift Valley fever virus (RVFV) are described. The assay was performed at a constant temperature (63 degrees C), with a real-time follow-up using a LightCycler and a double-stranded-DNA-intercalating fluorochrome. The assay is highly sensitive and comparable to real-time RT-PCR, with a detection limit of approximately 10 RNA copies per assay. However, the RT-LAMP assay is much faster than traditional RT-PCR and generates results in <30 min for most diluted samples. The specificity of the primers was established using other, related arboviruses as well as virus-containing and virus-free sera. The RT-LAMP assay reported here is thus a valuable tool for the rapid detection of RVFV in field diagnostic laboratories.
Peyrefitte
Christophe N
CN
Unité de virologie tropicale, Institut de Médecine tropicale du Service de Santé des Armées, BP 46, 13 998 Marseille armées, France.
Boubis
Laetitia
L
Coudrier
Daniel
D
Bouloy
Michèle
M
Grandadam
Marc
M
Tolou
Hugues J
HJ
Plumet
Sébastien
S
D017931:DNA Primers / Q000235:genetics; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D012295:Rift Valley Fever / Q000175:diagnosis* / Q000821:virology; D012296:Rift Valley fever virus / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors
D017931:DNA Primers; D012367:RNA, Viral
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