title: Development of Conventional and Real-Time Quantitative PCR Assays for Diagnosis and Monitoring of Scabies.
abstract: Scabies 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.
authors.affiliation: Department of Microbiology, Research Centre for Infection and Immunology, Faculty of Medicine, The University of Hong Kong, Hong Kong.
authors.forename: Samson S Y
authors.initials: SS
authors.lastname: Wong
authors.affiliation: Department of Microbiology, Queen Mary Hospital, Hong Kong.
authors.forename: Rosana W S
authors.initials: RW
authors.lastname: Poon
authors.affiliation: Department of Pathology, United Christian Hospital, Hong Kong.
authors.forename: Sandy
authors.initials: S
authors.lastname: Chau
authors.affiliation: Department of Microbiology, Queen Mary Hospital, Hong Kong.
authors.forename: Sally C Y
authors.initials: SC
authors.lastname: Wong
authors.affiliation: Department of Microbiology, Research Centre for Infection and Immunology, Faculty of Medicine, The University of Hong Kong, Hong Kong.
authors.forename: Kelvin K W
authors.initials: KK
authors.lastname: To
authors.affiliation: Department of Microbiology, Queen Mary Hospital, Hong Kong.
authors.forename: Vincent C C
authors.initials: VC
authors.lastname: Cheng
authors.affiliation: Department of Pathology, United Christian Hospital, Hong Kong.
authors.forename: Kitty S C
authors.initials: KS
authors.lastname: Fung
authors.affiliation: Department of Microbiology, Research Centre for Infection and Immunology, Faculty of Medicine, The University of Hong Kong, Hong Kong kyyuen@hku.hk.
authors.forename: K Y
authors.initials: KY
authors.lastname: Yuen
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D016903:Drug Monitoring / Q000379:methods*; D003576:Electron Transport Complex IV / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012520:Sarcoptes scabiei / Q000235:genetics; D012532:Scabies / Q000175:diagnosis* / Q000188:drug therapy; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D003576:Electron Transport Complex IV