Fig 3 -
Sensitivity of LAMP (A and B) and PCR (C) detection with sample dilution experiment (Left) and mosquito pool sample experiment (Right). LAMP assay with ethidium bromide-stained gel (A) and HNB indicator (B) of a fold-dilution of an individual crude boiling Aedes aegypti (wAlbB-TH) sample (182 ng/μl, 260/280 = 1.90, 260/230 = 0.99) diluted 10−1–10−6 times (6.4 units of Bst 2.0 DNA polymerase, 65°C for 90 min and 80°C for 10 min, volume 10 μl). Polymerase chain reaction with wsp primers (691R and 183F) of the same dilution (C). (O) is a non-diluted original sample, (P) is Wolbachia trans-infected Ae. aegypti (wAlbB-TH), (N1) is wild-type Ae. aegypti (Aae-JJ), (N2) is 100 wild-type Ae. aegypti (Aae-JJ), (NTC) is no template control, (M) is Invitrogen 100 bp DNA Ladder. For mosquito pooled experiment, the ratio of 1/25 indicates the DNA extraction was performed with one Wolbachia trans-infected Ae. aegypti and 24 wild-type Ae. aegypti. Other pooled samples included 1/50, 1/75, and 1/100, which were equivalent to one wAlbB-TH mosquito in 49, 74, and 99 Aae-JJ, respectively.