Journal: Pathogens

Article Title: High-Sensitivity RT-LAMP for Molecular Detection of O’nyong-nyong ( Alphavirus onyong )

doi: 10.3390/pathogens13100892

Figure Lengend Snippet: Comparison of the limit of detection of RT-LAMP ONNV between two different reaction times and specificity of RT-LAMP ONNV . ( a ) Limit of detection for a 20 min reaction time. ( b ) Limit of detection for a 30 min reaction time. ( c ) Agarose gel electrophoresis analysis of the specificity of the assay to detect ONNV, when performed with RNA of CHIKV, MAYV, and SINV. M: Molecular marker—GeneRuler 1 kb Plus DNA Ladder (Invitrogen). NTC: Negative control. RNA: ONNV RNA extract.

Article Snippet: Additionally, three other alphaviruses—namely, CHIKV (strain S-27; laboratory stock), SINV (strain EfAr339; laboratory stock), and MAYV (strain TRVL-4675, laboratory stock recently prepared after obtaining the virus through BEI Resources-NIAID-NIH)—all belonging to the Semliki Forest serological complex (all stored at −80 °C after replication on VeroE6 cells) were used for viral RNA extraction and included in the assay’s specificity evaluation.

Techniques: Comparison, Agarose Gel Electrophoresis, Marker, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Comparative Genomics, Infectivity and Cytopathogenicity of American Isolates of Zika Virus that Developed Persistent Infections in Human Embryonic Kidney (HEK293) Cells

doi: 10.3390/ijms20123035

Figure Lengend Snippet: Vero cell cultures infected using ZIKVs collected from the supernatants of acutely and persistently PRV and FLR strains of ZIKV-infected HEK293 cell cultures. Vero control cells as well as Vero cells infected by ZIKVs (PRV_V, PRV_H, Zp_H, FLR_V, FLR_H, Zf_H) using cell culture supernatants (Vero/PRV, HEK293/PRV, HEK293_Zp, Vero/FLR, HEK293/FLR, HEK293_Zf) with the same genomic copy number were stained four days post infection with True Blue after immunoblotting with Anti-Flavivirus Group antigen monoclonal antibody (Clone D1-4G2-4-15, BEI Resources) followed by goat anti-mouse IgG H&L (HRP). Separate sets of culture wells were stained in parallel with Crystal Violet to better visualize plaques formed by the infection of ZIKVs. Immunocytochemical staining showed that the infection foci produced by HEK293_Zp-generated persistent ZIKV (Zp_H) had poorly-formed lytic centers. HEK293_Zf-generated persistent ZIKV (Zf_H) produced significantly fewer infection foci. Moreover, the infection foci formed by Zf_H had no lytic centers.

Article Snippet: Viral inoculums were prepared from ZIKV stocks obtained from BEI Resources at multiplicity of infection (MOI) of 0.1 ± 0.01 according to the package description.

Techniques: Infection, Control, Cell Culture, Staining, Western Blot, Produced, Generated