TY - JOUR
T1 - Rapid detection of European orthobunyaviruses by reverse transcription loop-mediated isothermal amplification assays
AU - Camp, Jeremy V.
AU - Nowotny, Norbert
N1 - Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2016/10/1
Y1 - 2016/10/1
N2 - The development of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assays are described herein for the detection of two orthobunyaviruses (Bunyaviridae), which represent the two main serogroups found in mosquitoes in Central Europe. The RT-LAMP assays were optimized for the detection of Ťahyňa virus (a California encephalitis group virus found in Aedes sp or Ochlerotatus sp mosquitoes) and Batai virus (also called Čalovo virus, a Bunyamwera group virus found in Anopheles maculipennis s.l. mosquitoes) nucleic acid using endemic European virus isolates. The sensitivity of the RT-LAMP assays was determined to be comparable to that of conventional tests, with a limit of detection < 0.1 pfu per reaction. The assays can be performed in 60 min under isothermal conditions using very simple equipment. Furthermore, it was possible to proceed with the assays without nucleic acid extraction, albeit at a 100-fold loss of sensitivity. The RT-LAMP assays are a sensitive, cost-efficient method for both arbovirus surveillance as well as diagnostic laboratories to detect the presence of these endemic orthobunyaviruses.
AB - The development of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assays are described herein for the detection of two orthobunyaviruses (Bunyaviridae), which represent the two main serogroups found in mosquitoes in Central Europe. The RT-LAMP assays were optimized for the detection of Ťahyňa virus (a California encephalitis group virus found in Aedes sp or Ochlerotatus sp mosquitoes) and Batai virus (also called Čalovo virus, a Bunyamwera group virus found in Anopheles maculipennis s.l. mosquitoes) nucleic acid using endemic European virus isolates. The sensitivity of the RT-LAMP assays was determined to be comparable to that of conventional tests, with a limit of detection < 0.1 pfu per reaction. The assays can be performed in 60 min under isothermal conditions using very simple equipment. Furthermore, it was possible to proceed with the assays without nucleic acid extraction, albeit at a 100-fold loss of sensitivity. The RT-LAMP assays are a sensitive, cost-efficient method for both arbovirus surveillance as well as diagnostic laboratories to detect the presence of these endemic orthobunyaviruses.
KW - Arbovirus
KW - Bunyaviridae
KW - Loop-mediated isothermal amplification
KW - Mosquito
KW - Orthobunyavirus
KW - RT-LAMP
UR - http://www.scopus.com/inward/record.url?scp=84981297739&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84981297739&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2016.08.001
DO - 10.1016/j.jviromet.2016.08.001
M3 - Article
C2 - 27491341
AN - SCOPUS:84981297739
SN - 0166-0934
VL - 236
SP - 252
EP - 257
JO - Journal of Virological Methods
JF - Journal of Virological Methods
ER -