TY - JOUR
T1 - Optimization of DNA extraction from field-collected mammalian whole blood on filter paper for Trypanosoma cruzi (Chagas disease) detection
AU - Gulas-Wroblewski, Bonnie E.
AU - Kairis, Rebecca B.
AU - Gorchakov, Rodion Viktorovich
AU - Wheless, Anna
AU - Murray, Kristy O.
N1 - KAUST Repository Item: Exported on 2021-08-31
Acknowledgements: This research was funded in part from NIH/NIAID, grant number R03AI123650.
PY - 2021/8/17
Y1 - 2021/8/17
N2 - Blood filter paper strips are cost-effective materials used to store body fluid specimens under challenging field conditions, extending the reach of zoonotic pathogen surveillance and research. We describe an optimized procedure for the extraction of parasite DNA from whole blood (WB) stored on Type I Advantec Nobuto strips from both experimentally spiked and field-collected specimens from canine and skunks, respectively. When comparing two commercial kits for extraction, Qiagen’s DNeasy Blood & Tissue Kit performed best for the detection of parasite DNA by PCR from Trypanosoma cruzi-spiked canine WB samples on Nobuto strips. To further optimize recovery of β-actin from field-collected skunk WB archived on Nobuto strips, we modified the extraction procedures for the Qiagen kit with a 90 °C incubation step and extended incubation post-addition of proteinase K, a method subsequently employed to identify a T. cruzi infection in one of the skunks. Using this optimized extraction method can efficaciously increase the accuracy and precision of future molecular epidemiologic investigations targeting neglected tropical diseases in field-collected WB specimens on filter strips.
AB - Blood filter paper strips are cost-effective materials used to store body fluid specimens under challenging field conditions, extending the reach of zoonotic pathogen surveillance and research. We describe an optimized procedure for the extraction of parasite DNA from whole blood (WB) stored on Type I Advantec Nobuto strips from both experimentally spiked and field-collected specimens from canine and skunks, respectively. When comparing two commercial kits for extraction, Qiagen’s DNeasy Blood & Tissue Kit performed best for the detection of parasite DNA by PCR from Trypanosoma cruzi-spiked canine WB samples on Nobuto strips. To further optimize recovery of β-actin from field-collected skunk WB archived on Nobuto strips, we modified the extraction procedures for the Qiagen kit with a 90 °C incubation step and extended incubation post-addition of proteinase K, a method subsequently employed to identify a T. cruzi infection in one of the skunks. Using this optimized extraction method can efficaciously increase the accuracy and precision of future molecular epidemiologic investigations targeting neglected tropical diseases in field-collected WB specimens on filter strips.
UR - http://hdl.handle.net/10754/670836
UR - https://www.mdpi.com/2076-0817/10/8/1040
UR - http://www.scopus.com/inward/record.url?scp=85113218065&partnerID=8YFLogxK
U2 - 10.3390/pathogens10081040
DO - 10.3390/pathogens10081040
M3 - Article
C2 - 34451504
SN - 2076-0817
VL - 10
SP - 1040
JO - Pathogens
JF - Pathogens
IS - 8
ER -
