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Fig. 1 | BMC Research Notes

Fig. 1

From: Examining human paragonimiasis as a differential diagnosis to tuberculosis in The Gambia

Fig. 1

Assay validation electrophoresis gels. a To estimate the assay’s limit of detection of pure DNA, PCR was performed on a dilution series of plasmid positive control (1000, 100, 10, 5.0, 1.0, 0.5 and 0.1 pg/µl). Amplification was seen in all dilutions for P. africanus and P. uterobilateralis. Dilutions ranged from 1.0 ng/µl (dilution 1) to 0.1 pg/µl (dilution 7). Samples were run on the same gel with P. africanus and P. uterobilateralis samples, and a no template control (“NTC”). b To estimate the assay’s limit of detection of DNA in sputum, PCR was performed using DNA extracted from sputum spiked with positive control DNA. 12 sputum concentrates were spiked using the same dilution series as in (a). Samples were run on the same gel, with P. africanus and P. uterobilateralis samples, with plasmid positive control (“+ve cont”) and no template control (“NTC”). c Sensitivity of the PW-F/PAU-B primer set to the two West African target Paragonimus species was compared to P. westermani in a dilution series of pure plasmid positive control (1000, 100, 10 and 5.0 pg/µl). d Specificity of PW-F/PAU-B (Series A) designed for P. africanus and P. uterobilateralis were compared to that of primers PW-F/PW-B (Series B) for P. westermani. DNA from known positive patient samples infected with related worm species to Paragonimus were tested, including, (1) Schistosoma mansoni, (2) Schistosoma haematobium, (3) Necator americanus, (4) Opisthorchis spp. Amplification of a large sized product (800 base pairs) was seen for Opisthorchis spp. using the P. africanus/P. uterobilateralis primers. No unspecific amplification was seen using primers PW-F/PW-B

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