Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome.
Background The suboptimal sensitivity and specificity of available diagnostic methods for scabies hampers clinical management, trials of new therapies and epidemiologic studies. Additionally, parasitologic diagnosis by microscopic examination of skin scrapings requires sample collection with a sharp...
Published in: | PLOS Neglected Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:515d090e2c6947048554f98dbe118ace 2023-05-15T15:16:43+02:00 Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. Lena Chng Deborah C Holt Matt Field Joshua R Francis Dev Tilakaratne Milou H Dekkers Greg Robinson Kate Mounsey Rebecca Pavlos Asha C Bowen Katja Fischer Anthony T Papenfuss Robin B Gasser Pasi K Korhonen Bart J Currie James S McCarthy Cielo Pasay 2021-02-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0009149 https://doaj.org/article/515d090e2c6947048554f98dbe118ace EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0009149 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009149 https://doaj.org/article/515d090e2c6947048554f98dbe118ace PLoS Neglected Tropical Diseases, Vol 15, Iss 2, p e0009149 (2021) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2021 ftdoajarticles https://doi.org/10.1371/journal.pntd.0009149 2022-12-31T05:04:46Z Background The suboptimal sensitivity and specificity of available diagnostic methods for scabies hampers clinical management, trials of new therapies and epidemiologic studies. Additionally, parasitologic diagnosis by microscopic examination of skin scrapings requires sample collection with a sharp scalpel blade, causing discomfort to patients and difficulty in children. Polymerase chain reaction (PCR)-based diagnostic assays, combined with non-invasive sampling methods, represent an attractive approach. In this study, we aimed to develop a real-time probe-based PCR test for scabies, test a non-invasive sampling method and evaluate its diagnostic performance in two clinical settings. Methodology/principal findings High copy-number repetitive DNA elements were identified in draft Sarcoptes scabiei genome sequences and used as assay targets for diagnostic PCR. Two suitable repetitive DNA sequences, a 375 base pair microsatellite (SSR5) and a 606 base pair long tandem repeat (SSR6), were identified. Diagnostic sensitivity and specificity were tested using relevant positive and negative control materials and compared to a published assay targeting the mitochondrial cox1 gene. Both assays were positive at a 1:100 dilution of DNA from a single mite; no amplification was observed in DNA from samples from 19 patients with other skin conditions nor from house dust, sheep or dog mites, head and body lice or from six common skin bacterial and fungal species. Moderate sensitivity of the assays was achieved in a pilot study, detecting 5/7 (71.4% [95% CI: 29.0% - 96.3%]) of clinically diagnosed untreated scabies patients). Greater sensitivity was observed in samples collected by FLOQ swabs compared to skin scrapings. Conclusions/significance This newly developed qPCR assay, combined with the use of an alternative non-invasive swab sampling technique offers the possibility of enhanced diagnosis of scabies. Further studies will be required to better define the diagnostic performance of these tests. Article in Journal/Newspaper Arctic Mite Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 15 2 e0009149 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Lena Chng Deborah C Holt Matt Field Joshua R Francis Dev Tilakaratne Milou H Dekkers Greg Robinson Kate Mounsey Rebecca Pavlos Asha C Bowen Katja Fischer Anthony T Papenfuss Robin B Gasser Pasi K Korhonen Bart J Currie James S McCarthy Cielo Pasay Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Background The suboptimal sensitivity and specificity of available diagnostic methods for scabies hampers clinical management, trials of new therapies and epidemiologic studies. Additionally, parasitologic diagnosis by microscopic examination of skin scrapings requires sample collection with a sharp scalpel blade, causing discomfort to patients and difficulty in children. Polymerase chain reaction (PCR)-based diagnostic assays, combined with non-invasive sampling methods, represent an attractive approach. In this study, we aimed to develop a real-time probe-based PCR test for scabies, test a non-invasive sampling method and evaluate its diagnostic performance in two clinical settings. Methodology/principal findings High copy-number repetitive DNA elements were identified in draft Sarcoptes scabiei genome sequences and used as assay targets for diagnostic PCR. Two suitable repetitive DNA sequences, a 375 base pair microsatellite (SSR5) and a 606 base pair long tandem repeat (SSR6), were identified. Diagnostic sensitivity and specificity were tested using relevant positive and negative control materials and compared to a published assay targeting the mitochondrial cox1 gene. Both assays were positive at a 1:100 dilution of DNA from a single mite; no amplification was observed in DNA from samples from 19 patients with other skin conditions nor from house dust, sheep or dog mites, head and body lice or from six common skin bacterial and fungal species. Moderate sensitivity of the assays was achieved in a pilot study, detecting 5/7 (71.4% [95% CI: 29.0% - 96.3%]) of clinically diagnosed untreated scabies patients). Greater sensitivity was observed in samples collected by FLOQ swabs compared to skin scrapings. Conclusions/significance This newly developed qPCR assay, combined with the use of an alternative non-invasive swab sampling technique offers the possibility of enhanced diagnosis of scabies. Further studies will be required to better define the diagnostic performance of these tests. |
format |
Article in Journal/Newspaper |
author |
Lena Chng Deborah C Holt Matt Field Joshua R Francis Dev Tilakaratne Milou H Dekkers Greg Robinson Kate Mounsey Rebecca Pavlos Asha C Bowen Katja Fischer Anthony T Papenfuss Robin B Gasser Pasi K Korhonen Bart J Currie James S McCarthy Cielo Pasay |
author_facet |
Lena Chng Deborah C Holt Matt Field Joshua R Francis Dev Tilakaratne Milou H Dekkers Greg Robinson Kate Mounsey Rebecca Pavlos Asha C Bowen Katja Fischer Anthony T Papenfuss Robin B Gasser Pasi K Korhonen Bart J Currie James S McCarthy Cielo Pasay |
author_sort |
Lena Chng |
title |
Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. |
title_short |
Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. |
title_full |
Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. |
title_fullStr |
Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. |
title_full_unstemmed |
Molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the Sarcoptes scabiei genome. |
title_sort |
molecular diagnosis of scabies using a novel probe-based polymerase chain reaction assay targeting high-copy number repetitive sequences in the sarcoptes scabiei genome. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2021 |
url |
https://doi.org/10.1371/journal.pntd.0009149 https://doaj.org/article/515d090e2c6947048554f98dbe118ace |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic Mite |
genre_facet |
Arctic Mite |
op_source |
PLoS Neglected Tropical Diseases, Vol 15, Iss 2, p e0009149 (2021) |
op_relation |
https://doi.org/10.1371/journal.pntd.0009149 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009149 https://doaj.org/article/515d090e2c6947048554f98dbe118ace |
op_doi |
https://doi.org/10.1371/journal.pntd.0009149 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
15 |
container_issue |
2 |
container_start_page |
e0009149 |
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1766347011876978688 |