AC-ELISA and RT-PCR assays for the diagnosis of triatoma virus (TrV) in triatomines (Hemiptera: Reduviidae) species.
Triatoma virus (TrV) is the only entomopathogenic virus found in triatomines. TrV replicates in cells of the midgut epithelium of triatomines, causing a high mortality rate and delayed development of the infected insect. In this work, we report an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and a reverse transcription-polymerase chain reaction (RT-PCR) assay for detection of TrV infection. For antiserum production, rabbits and hens where inoculated with purified TrV. Antiserum reactivity was checked by immunodiffusion, and its specificity was confirmed by western blot and AC-ELISA. Totally 90 fecal samples from T. infestans were analysed. AC-ELISA and RT-PCR results correlated well with transmission electron microscopy (EM) observations, which are considered the gold standard, with Kappa values of 0.73 for AC-ELISA and 0.93 for RT-PCR when compared with EM. Applications and complementary uses of the two techniques reported in this work are discussed.
Marti
Gerardo A
GA
Centro Regional de Investigaciones Científicas y Transferencias Tecnológicas La Rioja (CRILAR, CONICET) Entre Ríos and Mendoza, 5310, Anillaco, La Rioja, Argentina. gmarti@crilar-conicet.com.ar
González
Ester T
ET
García
Juan J
JJ
Viguera
Ana R
AR
Guérin
Diego M A
DM
Echeverría
María G
MG
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006430:Hemiptera / Q000821:virology; D007118:Immunoassay; D007304:Insect Viruses / Q000302:isolation & purification / Q000472:pathogenicity; D046529:Microscopy, Electron, Transmission; D010849:Picornaviridae / Q000302:isolation & purification / Q000502:physiology / Q000648:ultrastructure*; D010850:Picornaviridae Infections / Q000175:diagnosis* / Q000276:immunology; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014224:Triatoma / Q000821:virology*
D000914:Antibodies, Viral