Fig 1.
Schematic representation of the optimization steps of reactions in portable equipment, and the development of the simplified extraction protocol, including the categories of sample involved in each step.
(Credit: Created by the author in BioRender.com).
Table 1.
Ct results and avarage Cts from evaluation of lysis solutions for simplified DNA extraction protocol, based on mammalian 18S rRNA target detection by qPCR in swine skin using QS-5 equipment.
Protocol numbers are summarized as 1—Guanidine (2 M) and Urea (2 M); 2 –Urea (2 M); 3 –Guanidine (3 M) and Urea (4 M); 4 –Guanidine (4 M); 5 –Guanidine (5 M); 6 –NH4OH (0.06 M); NC–Negative control.
Fig 2.
Schematic representation of the simplified DNA extraction protocol for Mycobacterium leprae from clinical skin biopsy samples.
(Credit: Created by the author in BioRender.com).
Table 2.
Comparison of molecular diagnostic parameters in 53 clinical biopsies in evaluated simplified extraction protocol and commercial protocol kit across different platforms (Q3-Plus and Quantstudio 5).
PB: paucibacillary; MB: multibacillary.
Fig 3.
Analytical sensitivity of qPCR reactions for the targets 16S rRNA and RLEP of Mycobacterium leprae performed on the portable Q3-Plus equipment.
(A) LOD95% 16S rRNA; (B) LOD95% RLEP. (Created by the author).
Table 3.
Comparison of molecular diagnostic parameters in 95 clinical biopsies evaluated in commercial extraction protocol kit across different platforms (ABI7500, QuantStudio-5 and Q3-Plus platforms).
PB: paucibacillary; MB: multibacillary.