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posted on 13.05.2022, 17:43 by Parinda Thayanukul, Benchaporn Lertanantawong, Worachart Sirawaraporn, Surat Charasmongkolcharoen, Thanyarat Chaibun, Rattanalak Jittungdee, Pattamaporn Kittayapong

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.