Supplementary information
Supplementary figure 1. Exclusivity of the LAMP reaction performed in the device. The exclusivity of the LAMP reaction performed in the device was checked. One ng of P.falciparum 3D7 genomic DNA was used in a LAMP reaction for 60 minutes, as described in Materials and Methods. (A) The confirmation of LAMP amplification was done by SYBR Gold detection.
Diluted SYBR Gold nucleic acid stain was added to the LAMP products, and the color change was detected by visual examination. --: non-template control, NC Pf: negative control with P.
falciparum 3D7 genomic DNA and L. donovani primers. (B) The quantification of SYBR Gold fluorescence was done by measuring the fluorescence intensity of the samples by a detection device, designated as ADC values. The mean + SEM of 10 ADC values is plotted for each sample. (C) Electrophoresis of LAMP products on a 2.5% agarose gel. M: 100 bp DNA ladder.
Supplementary figure 2. Time-course analysis of LAMP reaction. A time-course analysis of the LAMP reaction performed in the device was carried out. One ng of L. Donovani Bob genomic DNA was used in a LAMP reaction for 15, 30, 45 and 60 minutes, as described in Materials and Methods. (A) The confirmation of LAMP amplification was done by SYBR Gold detection.
Diluted SYBR Gold nucleic acid stain was added to the LAMP products, and the color change was detected by visual examination. --: non-template control. (B) The quantification of SYBR Gold fluorescence was done by measuring the fluorescence intensity of the samples by a detection device, designated as ADC values. The mean + SEM of 10 ADC values is plotted for each sample. (C) Electrophoresis of LAMP products on a 2.5% agarose gel. M: 100 bp DNA ladder.
Supplementary figure 3. LAMP assay to detect L. donovani DNA in non-endemic control (NEC) samples. One hundred pg of DNA from 15 NEC samples were used in a LAMP reaction for 60 minutes, as described in Materials and Methods. (A) The confirmation of LAMP amplification was done by SYBR Gold detection. Diluted SYBR Gold nucleic acid stain was added to the LAMP products, and the color change was detected by visual examination. --: non- template control. (B) The quantification of SYBR Gold fluorescence was done by measuring the fluorescence intensity of the samples by a detection device, designated as ADC values. The mean + SEM of 10 ADC values is plotted for each sample. (C) Electrophoresis of LAMP products on a 2.5% agarose gel. M: 100 bp DNA ladder.
Supplementary figure 4. LAMP assay to detect L. donovani DNA in leprosy patient samples.
One hundred pg of DNA from 10 leprosy patient samples were used in a LAMP reaction for 60 minutes, as described in Materials and Methods. (A) The confirmation of LAMP amplification was done by SYBR Gold detection. Diluted SYBR Gold nucleic acid stain was added to the LAMP products, and the color change was detected by visual examination. --: non-template control. (B) The quantification of SYBR Gold fluorescence was done by measuring the fluorescence intensity of the samples by a detection device, designated as ADC values. The mean + SEM of 10 ADC values is plotted for each sample. (C) Electrophoresis of LAMP products on a 2.5% agarose gel. M: 100 bp DNA ladder.
Supplementary figure 5. LAMP assay to detect L. donovani DNA in post-treatment VL patient samples. One hundred pg of DNA from 5 post-treatment VL samples were used in a LAMP reaction for 60 minutes, as described in Materials and Methods. (A) The confirmation of LAMP 1
amplification was done by SYBR Gold detection. Diluted SYBR Gold nucleic acid stain was added to the LAMP products, and the color change was detected by visual examination. --: non- template control. (B) The quantification of SYBR Gold fluorescence was done by measuring the fluorescence intensity of the samples by a detection device, designated as ADC values. The mean + SEM of 10 ADC values is plotted for each sample. (C) Electrophoresis of LAMP products on a 2.5% agarose gel. M: 100 bp DNA ladder.
2