Joanna Dąbrowska (Poland)
National Veterinary Research Institute
(NVRI)
Joanna Dąbrowska is a scientist who works in the National Veterinary Research Institute in Poland where she is conducting studies in the field of veterinary parasitology. Her main subject of interest are protozoan parasites. Joanna focuses on identification of parasites with the newest molecular tools. She developed LAMP and real time PCR as basic diagnostic tests for T. foetus detection from clinical samples of susceptible animals. Furthermore, during her 1 year scholarship in Switzerland she was involved in the Next Generation Sequencing of T.foetus in collaboration with Institute of Parasitology from University of Bern. Recently, she works on sequencing of parasites with nanopore technology. Furthermore, she is conducting the studies with proteomic and transcriptomic analysis of of parasites.

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03. Parasites of domestic and wild animals

PRE-RECORDED: COMPARISON OF FOUR COMMERCIAL METHODS FOR THE EXTRACTION DNA BY TWO MOLECULAR METHODS FOR TRITRICHOMONAS FOETUS DNA IDENTIFICATION IN SPIKED FELINE FECES (ID 1081)

Session Name
PA10-05 - OTHER STUDIES OF PARASITES OF DOMESTIC AND WILD ANIMALS - TOPIC: 3.6 OTHER STUDIES RELATED TO PARASITES OF DOMESTIC AND WILD ANIMALS
Session Type
03. Parasites of domestic and wild animals
Date
08/24/2022
Session Time
13:15 - 14:45
Room
Hall B4.M7+8
Presenter
Lecture Time
14:20 - 14:25
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Pre-Recorded Presentation
Onsite or Pre-Recorded
Pre-Recorded

Abstract

Introduction

Tritrichomonas foetus causes large bowel diarrhea in cats. Due to low specificity, it is difficult to identify a parasite only based on clinical signs. Therefore, molecular tests are currently used. However, the extraction of DNA from fecal samples can be quite challenging because of the presence of PCR inhibitors that are coextracted with DNA. In our study, four commercially available DNA extraction kits were compared in terms of their effect on the two molecular assays.

Methods

Fecal samples were spiked with an appropriate number of T.foetus cells (series of dilutions corresponded to 10000, 1000, 100, 10, 1 and 0.1 cells per sample, each variant in 6 replicates). DNA of T. foetus from feces was extracted using:(Q) QIAamp® DNA Stool Mini Kit (Qiagen Inc., Valencia, CA), (U) UltraClean Fecal DNA Kit (50 preps) (MO BIO, San Diego, CA), (S) Sherlock AX / 100 isolation (A&A Biotechnology, Gdynia, Poland) and (Z) ZR Fecal MiniPrep (Zymo Research, Irvine, CA). PCR and the in-house LAMP for the identification of T. foetus were used for the evaluation of isolation kits.

Results

The highest number of positive results were obtained by LAMP with Z (33.3%) in fecal samples spiked with equivalent to 0.1 cell. A lower number of positive results were found after extraction by Q in LAMP (66%) and PCR (33%) with 1 cell. Surprisingly, S has relatively high efficiency. LAMP was able to detect Tritrichomonas foetus cells on the level of spiking 10 cells with 16.6%.The least sensitive method was U which could only detect 100 cells (33%) by LAMP and 100 cells (16,6%) by PCR.

Conclusions

Our investigation showed the highest efficacy Z dedicated for fecal samples in T. foetus detection by LAMP. Furthermore, it is worth to noticed that the first-time efficacy of the extractions method was evaluated by LAMP.

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