Abstract
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.
Highlights
The parasitic protozoan Trypanosoma cruzi is the etiologic agent of Chagas disease, which is maintained in domestic, peridomestic, and sylvatic transmission cycles by a diversity of triatomine vectors and mammalian hosts [1]
Pathogens 2021, 10, 1040 performed those relying on Zymo Research kits for both whole blood (WB) and WB stored on Nobuto strips (Supplementary Table S1)
Differences between the mean Cq values generated by high spiking loads for WB and WB stored on Nobuto blood filter paper were lower for the Qiagen DNeasy Blood & Tissue Kit methods
Summary
The parasitic protozoan Trypanosoma cruzi is the etiologic agent of Chagas disease, which is maintained in domestic, peridomestic, and sylvatic transmission cycles by a diversity of triatomine vectors and mammalian hosts [1]. This neglected tropical parasite infects an estimated 6–7 million people across the Americas, making the zoonosis one of the most significant in terms of disease burden and public health importance in the western hemisphere [2]. Developing optimized protocols for T. cruzi DNA extraction from blood filter papers greatly expands the efficiency and effectiveness of field investigations into the molecular epidemiology and surveillance of Chagas disease among mammalian reservoirs
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