Validation of DESS as a DNA Preservation Method for the Detection of Strongyloides spp. in Canine Feces

• Main Authors: Meruyert Beknazarova, Shelby Millsteed, Gemma Robertson, Harriet Whiley, Kirstin Ross
• Format: Article
• Language: English
• Published: MDPI AG 2017-06-01
• Series: International Journal of Environmental Research and Public Health
• Subjects: Strongyloides stercoralis; Strongyloides ratti; Strongyloides; real-time PCR; DESS; DNA preservation; DNA degradation; canine feces
• Online Access: http://www.mdpi.com/1660-4601/14/6/624

Strongyloides stercoralis is a gastrointestinal parasitic nematode with a life cycle that includes free-living and parasitic forms. For both clinical (diagnostic) and environmental evaluation, it is important that we can detect Strongyloides spp. in both human and non-human fecal samples. Real-time PCR is the most feasible method for detecting the parasite in both clinical and environmental samples that have been preserved. However, one of the biggest challenges with PCR detection is DNA degradation during the postage time from rural and remote areas to the laboratory. This study included a laboratory assessment and field validation of DESS (dimethyl sulfoxide, disodium EDTA, and saturated NaCl) preservation of Strongyloides spp. DNA in fecal samples. The laboratory study investigated the capacity of 1:1 and 1:3 sample to DESS ratios to preserve Strongyloides ratti in spike canine feces. It was found that both ratios of DESS significantly prevented DNA degradation compared to the untreated sample. This method was then validated by applying it to the field-collected canine feces and detecting Strongyloides DNA using PCR. A total of 37 canine feces samples were collected and preserved in the 1:3 ratio (sample: DESS) and of these, 17 were positive for Strongyloides spp. The study shows that both 1:1 and 1:3 sample to DESS ratios were able to preserve the Strongyloides spp. DNA in canine feces samples stored at room temperature for up to 56 days. This DESS preservation method presents the most applicable and feasible method for the Strongyloides DNA preservation in field-collected feces.