Validation of DESS as a DNA Preservation Method for the Detection of Strongyloides spp. in Canine Feces
نویسندگان
چکیده
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.
منابع مشابه
Is the Baermann, an applicable method for detection of Strongyloides stercoralis?
Introduction: It is estimated that 100 - 300 million people in the world are infected with Strongyloides stercoralis. Although there are several laboratory diagnostic methods for detection of this parasite, there is still no gold standard diagnostic method. The need for an application test with high sensitivity and specificity is highly felt, especially in people with immune deficiency or organ...
متن کاملPolymerase chain reaction method for the rapid detection of virulent Shigella spp.
Bacillary dysentery, or shigellosis, is a disease of humans in which the colonic epithelium is invaded by bacteria and subjected to inflammatory destruction. The aim of this study was to develop a polymerase chain reaction(PCR) test for detection of virulent Shigella spp.. For this purpose, the primers were designed to amplify a 526-bp internal region of the Shigella spp. icsA gene, which encod...
متن کاملDetection of single Dactylogyrus spp. in DNA extracted from infected gill tissue of fishes using Polymerase Chain Reaction
Dactylogyrus spp. are monogenean worms found mostly as ectoparasites on the gills of several fish species, including carp and goldfish. These parasites are commonly detected by microscopic analysis of the gill lamellae, but this is time-consuming and technically difficult. In contrast to this conventional method, molecular techniques provide specific, sensitive and safe detection of parasites. ...
متن کاملStrong-LAMP: A LAMP Assay for Strongyloides spp. Detection in Stool and Urine Samples. Towards the Diagnosis of Human Strongyloidiasis Starting from a Rodent Model
BACKGROUND Strongyloides stercoralis, the chief causative agent of human strongyloidiasis, is a nematode globally distributed but mainly endemic in tropical and subtropical regions. Chronic infection is often clinically asymptomatic but it can result in severe hyperinfection syndrome or disseminated strongyloidiasis in immunocompromised patients. There is a great diversity of techniques used in...
متن کاملMolecular diagnosis of Mycoplasma spp. Arthritis by PCR
Background: Arthritis is one of the most common inflammatory diseases worldwide. It is characterized by symptoms such as systemic inflammation and autoantibody production. The molecular mechanisms in pathogenesis of arthritis are not fully understood. Studies show that many microorganisms, including Mycoplasmas, play a role in arthritis. The PCR method is a fast and accurate molecular method fo...
متن کامل