Comparison of Two DNA Extraction Methods and Two PCRs for Detection of <i>Echinococcus multilocularis</i> in the Stool Samples of Naturally Infected Red Foxes

(1) Background: Due to the increasing distribution of <i>Echinococcus multilocularis</i> infections in final hosts, epidemiological investigations are important for recognizing the spreading pattern of this parasite and also to estimate risk infection for humans. (2) Methods: Investigati...

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Main Authors: Katarzyna Skrzypek, Jacek Karamon, Małgorzata Samorek-Pieróg, Joanna Dąbrowska, Maciej Kochanowski, Jacek Sroka, Ewa Bilska-Zając, Tomasz Cencek
Format: Article
Language:English
Published: MDPI AG 2020-12-01
Series:Animals
Subjects:
Online Access:https://www.mdpi.com/2076-2615/10/12/2381
Description
Summary:(1) Background: Due to the increasing distribution of <i>Echinococcus multilocularis</i> infections in final hosts, epidemiological investigations are important for recognizing the spreading pattern of this parasite and also to estimate risk infection for humans. (2) Methods: Investigations were conducted with two commercial kits dedicated for DNA extraction from feces: ZR Fecal DNA Mini Prep (Zymo Research, Freiburg, Germany) and QIAamp FAST DNA Stool Mini Kit (Qiagen, Hilden, Germany) (marked as Z and Q), together with two common PCR protocols (nested PCR and multiplex PCR). The goal was to compare their efficiency in detecting the genetic material of <i>E. multilocularis</i> in the samples of feces. Stool samples from red foxes were collected in a highly endemic area in Poland. Sedimentation and counting technique (SCT) was used as a reference method. (3) Results: From 48 samples, 35 were positive in SCT. Further investigations showed that 40.0% of samples (from those with SCT positive result) after Z-DNA extraction and 45.7% after Q-DNA extraction gave positive results in nested PCR. In multiplex PCR, positive results were obtained in 54.3% of samples after Z isolation and 48.6% of samples after Q. Additionally, one sample that resulted in being negative in SCT gave a positive result in PCR. The number of worms detected in the intestines had no influence on PCR results. (4) Conclusions: Both of the extraction methods showed similar efficiency in DNA isolation and dealing with inhibitors; however, they showed relatively low sensitivity. This was probably caused by degradation of genetic material in the field-collected samples.
ISSN:2076-2615