Optimization of cationic (Q)-paper for detection of arboviruses in infected mosquitoes.
Previously (Glushakova et al. 2017), a cellulose-based cationic (Q) paper derivatized with quaternary ammonium groups was shown to be a convenient platform to collect, preserve, and store nucleic acids (NAs) derived from mosquito vectors infected with pathogens for surveillance. NAs bind electrostatically to Q-paper, but the quantity of NA bound depends on the paper's binding capacity. To optimize the original technology for mosquito surveillance, factors that affected NA absorbance on Q-paper were evaluated. Sixteen variations of Q-paper were prepared with modifications of the derivatizing reagents and derivatization temperature. The binding capacities of these variations were determined first with 1,3,5-benzenetricarboxylic (BTCA), then viral RNA (purified or in infected mosquito samples) was used for validation. For this, samples with Zika (ZIKV) and chikungunya (CHIKV) RNA or virus-infected Aedes aegypti mosquito bodies were applied to sixteen Q-paper variants. Washing the paper samples with water versus elution with aqueous salt (1 M) gave samples that were analyzed for viral RNA by a PCR-based direct Luminex hybridization assay. The comparison ranked the Q-paper binding capacities from the lowest to the highest. The Q-paper with the highest RNA binding capability was further validated with ZIKV- and CHIKV-infected mosquito saliva.
['Glushakova|Lyudmyla G|LG|', 'Alto|Barry W|BW|', 'Kim|Myong-Sang|MS|', 'Wiggins|Keenan|K|', 'Eastmond|Bradley|B|', 'Moussatche|Patricia|P|', 'Burkett-Cadena|Nathan D|ND|', 'Benner|Steven A|SA|']
D000330:Aedes D000818:Animals D001103:Arboviruses D004772:Entomology D010209:Paper D016133:Polymerase Chain Reaction D012367:RNA, Viral D013048:Specimen Handling D014773:Virology
Arboviruses Mosquito Cationic (Q) paper