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Article: Development of Conventional and Real-Time Quantitative PCR Assays for Diagnosis and Monitoring of Scabies

TitleDevelopment of Conventional and Real-Time Quantitative PCR Assays for Diagnosis and Monitoring of Scabies
Authors
Issue Date2015
PublisherAmerican Society for Microbiology. The Journal's web site is located at http://jcm.asm.org/
Citation
Journal of Clinical Microbiology, 2015, v. 53, p. 2095-2102 How to Cite?
AbstractScabies remains the most prevalent, endemic, and neglected ectoparasitic infestation globally and can cause institutional outbreaks. The sensitivity of routine microscopy for demonstration of Sarcoptes scabiei mites or eggs in skin scrapings is only about 50%. Except for three studies using conventional or two-tube nested PCR on a small number of cases, no systematic study has been performed to improve the laboratory diagnosis of this important infection. We developed a conventional and a real-time quantitative PCR (qPCR) assay based on the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of S. scabiei. The cox1 gene is relatively well conserved, with its sequence having no high levels of similarity to the sequences of other human skin mites, pathogenic zoonotic mites, or common house dust mite species. This mitochondrial gene is also present in large quantities in arthropod cells, potentially improving the sensitivity of a PCR-based assay. In our study, both assays were specific and were more sensitive than microscopy in diagnosing scabies, with positive and negative predictive values of 100%. The S. scabiei DNA copy number in the microscopy-positive specimens was significantly higher than that in the microscopy-negative specimens (median S. scabiei DNA copy number, 3.604 versus 2.457 log10 copies per reaction; P = 0.0213). In the patient with crusted scabies, the qPCR assay performed on lesional skin swabs instead of scrapings revealed that the parasite DNA load took about 2 weeks to become negative after treatment. The utility of using lesional skin swabs as an alternative sample for diagnosis of scabies by PCR should be further evaluated.
Persistent Identifierhttp://hdl.handle.net/10722/217194
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorWong, SSY-
dc.contributor.authorPoon, RWS-
dc.contributor.authorChau, S-
dc.contributor.authorWong, SCY-
dc.contributor.authorTo, KKW-
dc.contributor.authorCheng, VCC-
dc.contributor.authorFung, KSC-
dc.contributor.authorYuen, KY-
dc.date.accessioned2015-09-18T05:51:38Z-
dc.date.available2015-09-18T05:51:38Z-
dc.date.issued2015-
dc.identifier.citationJournal of Clinical Microbiology, 2015, v. 53, p. 2095-2102-
dc.identifier.issn0095-1137-
dc.identifier.urihttp://hdl.handle.net/10722/217194-
dc.description.abstractScabies remains the most prevalent, endemic, and neglected ectoparasitic infestation globally and can cause institutional outbreaks. The sensitivity of routine microscopy for demonstration of Sarcoptes scabiei mites or eggs in skin scrapings is only about 50%. Except for three studies using conventional or two-tube nested PCR on a small number of cases, no systematic study has been performed to improve the laboratory diagnosis of this important infection. We developed a conventional and a real-time quantitative PCR (qPCR) assay based on the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of S. scabiei. The cox1 gene is relatively well conserved, with its sequence having no high levels of similarity to the sequences of other human skin mites, pathogenic zoonotic mites, or common house dust mite species. This mitochondrial gene is also present in large quantities in arthropod cells, potentially improving the sensitivity of a PCR-based assay. In our study, both assays were specific and were more sensitive than microscopy in diagnosing scabies, with positive and negative predictive values of 100%. The S. scabiei DNA copy number in the microscopy-positive specimens was significantly higher than that in the microscopy-negative specimens (median S. scabiei DNA copy number, 3.604 versus 2.457 log10 copies per reaction; P = 0.0213). In the patient with crusted scabies, the qPCR assay performed on lesional skin swabs instead of scrapings revealed that the parasite DNA load took about 2 weeks to become negative after treatment. The utility of using lesional skin swabs as an alternative sample for diagnosis of scabies by PCR should be further evaluated.-
dc.languageeng-
dc.publisherAmerican Society for Microbiology. The Journal's web site is located at http://jcm.asm.org/-
dc.relation.ispartofJournal of Clinical Microbiology-
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.-
dc.titleDevelopment of Conventional and Real-Time Quantitative PCR Assays for Diagnosis and Monitoring of Scabies-
dc.typeArticle-
dc.identifier.emailWong, SSY: samsonsy@hkucc.hku.hk-
dc.identifier.emailTo, KKW: kelvinto@hkucc.hku.hk-
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hk-
dc.identifier.authorityWong, SSY=rp00395-
dc.identifier.authorityTo, KKW=rp01384-
dc.identifier.authorityYuen, KY=rp00366-
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1128/JCM.00073-15-
dc.identifier.pmid25903566-
dc.identifier.pmcidPMC4473232-
dc.identifier.scopuseid_2-s2.0-84932628395-
dc.identifier.hkuros251568-
dc.identifier.volume53-
dc.identifier.spage2095-
dc.identifier.epage2102-
dc.identifier.isiWOS:000358287700015-
dc.publisher.placeUnited States-

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