Rapid identification of virus-carrying mosquitoes using reverse transcription-loop-mediated isothermal amplification.
Mosquitoes are critical vectors in many arboviral transmission cycles. Considering the increasing incidence of arboviral infections throughout the world, monitoring of vector populations for the presence of an arbovirus could be considered an important initial step of risk assessment to humans and animals. In response to this need, increased efforts to develop rapid and reliable diagnostic techniques have been undertaken; a single-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect virus in vector mosquitoes (Aedes aegypti) using the Flock House Virus (FHV) as a model. The robustness of the RT-LAMP reaction was revealed by its ability to detect FHV from an "all-in-one" template using whole mosquito bodies within 30min. Furthermore, RT-LAMP identified successfully a mosquito carrying just a single FHV particle, a level easily overlooked in conventional analysis such as plaque forming assays. These observations suggest that RT-LAMP is more reliable and useful for routine diagnosis of vector mosquitoes in regions where the prevalence of vector-borne diseases such as West Nile fever or dengue fever are common.
Perera
Namal
N
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Aonuma
Hiroka
H
Yoshimura
Aya
A
Teramoto
Tokiyasu
T
Iseki
Hiroshi
H
Nelson
Bryce
B
Igarashi
Ikuo
I
Yagi
Takeshi
T
Fukumoto
Shinya
S
Kanuka
Hirotaka
H
D000330:Aedes / Q000821:virology*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D005260:Female; D007304:Insect Viruses / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012328:RNA Viruses / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity; D014776:Virus Cultivation
D017931:DNA Primers; D012367:RNA, Viral