Detection of chikungunya viral RNA in mosquito bodies on cationic (Q) paper based on innovations in synthetic biology

Abstract

Chikungunya virus (CHIKV) represents a growing and global concern for public health that needs inexpensive and convenient methods to collect mosquitoes as potential carriers so that they can be preserved, stored and transported for later and/or remote analysis. Reported here is a cellulose-based paper, derivatized with quaternary ammonium groups (“Q-paper”) that meets these needs. In a series of tests, infected mosquito bodies were squashed directly on to Q-paper. Aqueous ammonia was then added on the mosquito bodies to release viral RNA that adsorbed on the cationic surface via electrostatic interactions. The samples were then stored (frozen) or transported. For analysis, the CHIKV nucleic acids were eluted from the Q-paper and PCR amplified in a workflow, previously developed, that also exploited two nucleic acid innovations, (“artificially expanded genetic information systems”, AEGIS, and “self-avoiding molecular recognition systems”, SAMRS). The amplicons were then analyzed by a Luminex hybridization assay. This procedure detected CHIKV RNA, if present, in each infected mosquito sample, but not in non-infected counterparts or ddH2O samples washes, with testing one week or ten months after sample collection.

Transmission potential of Mayaro virus in Florida Aedes aegypti and Aedes albopictus mosquitoes

“…Mosquitoes were tested for their potential to transmit MAYV at four time‐points after their ingestion of infected blood (3, 6, 9 and 12 days post‐feeding) by two different methods of collecting expectorated saliva. Equal numbers from both species and time intervals were divided up to be tested by collection of saliva in capillary tubes and cationic (Q) papers (Glushakova et al ., ). Mosquitoes chosen for Q‐paper assays were placed in 37‐mL plastic tubes (8 cm in height, 3 cm in diameter) 1 day before the time‐point at which the test was to be conducted.…”

Multiplexed kit based on Luminex technology and achievements in synthetic biology discriminates Zika, chikungunya, and dengue viruses in mosquitoes

“…Larvae were reared at an approximate density of 150 larvae/L water in plastic trays (25 cm × 30 cm × 5 cm) with water (900 mL) and larval food (0.4 g) consisting of equal amounts of brewer’s yeast and liver powder (by weight) at hatching, and supplemented again with the same amount of food 3–4 days later. Mosquitoes were maintained as described in [34] and allowed to feed on chickens at the Florida Medical Entomology Laboratory once per week to propagate eggs (IACUC protocol no. 201507682).…”

“…Infected mosquitoes and saliva were also positively assessed on the surface of a cationic (Q)-paper [34, 35] that bound mosquito and viral nucleic acids via electrostatic interactions.

Fig. 1Adapted from [32, 34]. Left panels : Overview of the Luminex xMAP DHAs protocol developed to detect DENV, CHIKV, and ZIKV RNAs.

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“…Adapted from [32, 34]. Left panels : Overview of the Luminex xMAP DHAs protocol developed to detect DENV, CHIKV, and ZIKV RNAs.…”

Interaksi Molekuler Amonium Hidroksida

“…Jika napas sulit, berikan oksigen. Dapatkan bantuan medis (96,(193)(194)(195)(196)(197)(198)(199)(200)(201)(202)(203)(204)(205)(206)(207)(208)(209)(210)(211) . Catatan dokter : Ssetelah paparan inhalasi, amati selama 24 s.d 72 jam sebagai edema paru mungkin tertunda…”