Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis.
Background Developing more sensitive methods for the diagnosis of echinococcosis is essential. In this study PCR assay for sensitive detection of specific cell-free DNA (cfDNA) of Echinococcus granulosus sensu lato in the sera of the sheep naturally infected with echinococcosis was investigated. Met...
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ftdoajarticles:oai:doaj.org/article:8d67a12e29dc475aa174a741dc23574d 2023-12-03T10:18:11+01:00 Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. Mahboubeh Hadipour Hossein Yousofi Darani Hamid Talebzadeh Mohammad Eslamian Shima Aboutalebian Majid Fasihi Harandi Hossein Mirhendi 2023-10-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0011715 https://doaj.org/article/8d67a12e29dc475aa174a741dc23574d EN eng Public Library of Science (PLoS) https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011715&type=printable https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0011715 https://doaj.org/article/8d67a12e29dc475aa174a741dc23574d PLoS Neglected Tropical Diseases, Vol 17, Iss 10, p e0011715 (2023) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2023 ftdoajarticles https://doi.org/10.1371/journal.pntd.0011715 2023-11-05T01:35:31Z Background Developing more sensitive methods for the diagnosis of echinococcosis is essential. In this study PCR assay for sensitive detection of specific cell-free DNA (cfDNA) of Echinococcus granulosus sensu lato in the sera of the sheep naturally infected with echinococcosis was investigated. Methods To extract cfDNA from 35 infected sheep, the modified phenol-chloroform method was used for two different volumes (0.5 and 2 ml) of serum samples. From each extracted sample, two DNA volumes (5 and 10 μl) were amplified using both standard PCR and semi-nested PCR targeting NADH dehydrogenase subunit I. Results Standard and semi-nested PCR on 0.5 ml of serum samples detected Echinococcus DNA in 8 and 12 out of 35 sheep, respectively; however, using 2 ml of serum samples, they detected 24 and 27 samples. By increasing the volume of template DNA, the PCRs could detect 29 and 33 out of 35 samples. The results were confirmed by sequencing of randomly selected PCR amplicons and comparing them with GenBank databases. Conclusions Larger volumes of serum for DNA extraction, greater volumes of DNA template for PCR, and employing a semi-nested PCR protocol, increased the sensitivity of PCR to 95%. This approach can also be applied to the diagnosis of echinococcosis in humans. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 17 10 e0011715 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Mahboubeh Hadipour Hossein Yousofi Darani Hamid Talebzadeh Mohammad Eslamian Shima Aboutalebian Majid Fasihi Harandi Hossein Mirhendi Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Background Developing more sensitive methods for the diagnosis of echinococcosis is essential. In this study PCR assay for sensitive detection of specific cell-free DNA (cfDNA) of Echinococcus granulosus sensu lato in the sera of the sheep naturally infected with echinococcosis was investigated. Methods To extract cfDNA from 35 infected sheep, the modified phenol-chloroform method was used for two different volumes (0.5 and 2 ml) of serum samples. From each extracted sample, two DNA volumes (5 and 10 μl) were amplified using both standard PCR and semi-nested PCR targeting NADH dehydrogenase subunit I. Results Standard and semi-nested PCR on 0.5 ml of serum samples detected Echinococcus DNA in 8 and 12 out of 35 sheep, respectively; however, using 2 ml of serum samples, they detected 24 and 27 samples. By increasing the volume of template DNA, the PCRs could detect 29 and 33 out of 35 samples. The results were confirmed by sequencing of randomly selected PCR amplicons and comparing them with GenBank databases. Conclusions Larger volumes of serum for DNA extraction, greater volumes of DNA template for PCR, and employing a semi-nested PCR protocol, increased the sensitivity of PCR to 95%. This approach can also be applied to the diagnosis of echinococcosis in humans. |
format |
Article in Journal/Newspaper |
author |
Mahboubeh Hadipour Hossein Yousofi Darani Hamid Talebzadeh Mohammad Eslamian Shima Aboutalebian Majid Fasihi Harandi Hossein Mirhendi |
author_facet |
Mahboubeh Hadipour Hossein Yousofi Darani Hamid Talebzadeh Mohammad Eslamian Shima Aboutalebian Majid Fasihi Harandi Hossein Mirhendi |
author_sort |
Mahboubeh Hadipour |
title |
Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. |
title_short |
Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. |
title_full |
Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. |
title_fullStr |
Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. |
title_full_unstemmed |
Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis. |
title_sort |
sensitive detection of specific cell-free dna in serum samples from sheep with cystic echinococcosis. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2023 |
url |
https://doi.org/10.1371/journal.pntd.0011715 https://doaj.org/article/8d67a12e29dc475aa174a741dc23574d |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 17, Iss 10, p e0011715 (2023) |
op_relation |
https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011715&type=printable https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0011715 https://doaj.org/article/8d67a12e29dc475aa174a741dc23574d |
op_doi |
https://doi.org/10.1371/journal.pntd.0011715 |
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PLOS Neglected Tropical Diseases |
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17 |
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e0011715 |
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