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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| Format: | Article |
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MDPI AG
2020-12-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/10/12/2381 |
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| author | Katarzyna Skrzypek Jacek Karamon Małgorzata Samorek-Pieróg Joanna Dąbrowska Maciej Kochanowski Jacek Sroka Ewa Bilska-Zając Tomasz Cencek |
| author_facet | Katarzyna Skrzypek Jacek Karamon Małgorzata Samorek-Pieróg Joanna Dąbrowska Maciej Kochanowski Jacek Sroka Ewa Bilska-Zając Tomasz Cencek |
| author_sort | Katarzyna Skrzypek |
| collection | DOAJ |
| description | (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. |
| first_indexed | 2024-03-10T14:07:49Z |
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| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T14:07:49Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-746445d7629a488a9911232bbec6278f2023-11-21T00:28:38ZengMDPI AGAnimals2076-26152020-12-011012238110.3390/ani10122381Comparison of Two DNA Extraction Methods and Two PCRs for Detection of <i>Echinococcus multilocularis</i> in the Stool Samples of Naturally Infected Red FoxesKatarzyna Skrzypek0Jacek Karamon1Małgorzata Samorek-Pieróg2Joanna Dąbrowska3Maciej Kochanowski4Jacek Sroka5Ewa Bilska-Zając6Tomasz Cencek7Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, PolandDepartment of Parasitology and Invasive Diseases, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, Poland(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.https://www.mdpi.com/2076-2615/10/12/2381<i>Echinococcus multilocularis</i>PCRDNA extractionfeces |
| spellingShingle | Katarzyna Skrzypek Jacek Karamon Małgorzata Samorek-Pieróg Joanna Dąbrowska Maciej Kochanowski Jacek Sroka Ewa Bilska-Zając Tomasz Cencek 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 Animals <i>Echinococcus multilocularis</i> PCR DNA extraction feces |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | <i>Echinococcus multilocularis</i> PCR DNA extraction feces |
| url | https://www.mdpi.com/2076-2615/10/12/2381 |
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