Figure 1.

Optimization of the RT-LAMP reaction for the detection of HCV. Visual detection of RT-LAMP products by pre-adding HNB based on different conditions: (A) incubation time, (B) effect of loop primers, (C) Mg²⁺ concentration, (D) ratio between the inner and outer primers, and (E) HNB concentration. HCV = hepatitis C virus; NTC = no template control. In Figure 1C, the NTC reaction tubes turned sky blue due to a reduction in Mg²⁺ concentration, even though the amplification process had not yet been started. Figure 1D shows that an increase in inner primers may result in a non-specific amplification, reducing the Mg²⁺ concentration in the reaction mixture and then presenting as sky blue, even though in NTC tubes.