Fig 1.
CHIKV-CZC-LAMP detected <50 PFU CHIKV.
(A) CHIKV-CZC-LAMP reaction tubes (upper column) and the equipment used for the CHIKV-CZC-LAMP reaction and detection (lower column). CHIKV-CZC-LAMP reaction tubes have all-in-one dried reagents that can be stored at room temperature. The amplifications were performed at 63°C, and the positive yellow fluorescent signals were observed with the LED LAMP reaction detector. (B) Representative view of CHIKV-CZC-LAMP results. The decreasing concentration of CHIKV in 1 μl of RNA or 1 μl RNA spiked with 1 μl of healthy human blood were tested. As a negative control, blood without CHIKV RNA was used. Upper panel shows fluorescent signals detected by 505 nm blue green LED detector. Lower panel are showing the view of naked eyes. (C) The time of positivity measured by real-time RT-LAMP detected with FAM channel, and the calculated sensitivity for serially diluted CHIKV RNA with or without blood were shown. The same RNA samples were used for qRT-PCR and the calculated Ct value were shown.
Table 1.
RT-LAMP results using RNA and serum from chikungunya patient and endemic healthy control in Rio de Janeiro, Brazil.
Fig 2.
Phylogenetic tree of sequences from LAMP-Nanopore analysis.
NJ tree based on 117 bp sequences determined by CHIKV-CZC-LAMP-MinION sequencing (bold) analysis, and 29 sequences from the GenBank were constructed. The nucleotides at the 27 SNPs within the amplified region were also indicated.
Table 2.
MinION sequence summary.