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. 2015 Nov 14:8:587.
doi: 10.1186/s13071-015-1204-8.

Universal conventional and real-time PCR diagnosis tools for Sarcoptes scabiei

Samer Angelone-Alasaad  1   2 , AnnaRita Molinar Min  3 , Mario Pasquetti  4 , Abdulaziz N Alagaili  5 , Stefano D'Amelio  6 , Federica Berrilli  7 , Vincent Obanda  8 , Mohamed A Gebely  9 , Ramón C Soriguer  10 , Luca Rossi  11
Affiliations

Universal conventional and real-time PCR diagnosis tools for Sarcoptes scabiei

Samer Angelone-Alasaad et al. Parasit Vectors. .

Erratum in

Abstract

Background: The mite Sarcoptes scabiei has a known host-range of over 100 mammal species including humans. One of the prime objectives of the Sarcoptes-World Molecular Network (WMN) is to design and develop universal Sarcoptes PCR-based diagnosis methods.

Methods: We describe here for the first time two universal mitochondrial-based diagnosis methods: (i) conventional end-point PCR and (ii) TaqMan real-time PCR. The design of both of these universal diagnosis methods was based on Sarcoptes samples collected from 23 host species in 14 countries.

Results: These methods, based on skin scrapings, were successfully used to etiologically confirm the diagnosis of different clinical degrees of sarcoptic mange in 48 animals belonging to six species. These universal PCR-based diagnosis methods are highly specific, technically sensitive and simple, and are based on the amplification of 135 bp from the Mitochondrial 16S rDNA. The method based on TaqMan real-time qPCR was more sensitive than the conventional end-point PCR.

Conclusions: Two universal PCR-based diagnosis methods for S. scabiei were successfully designed and applied; one based on conventional end-point PCR and the other on TaqMan real-time PCR. We recommend further testing and the application of these new universal methods worldwide.

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Figures

Fig. 1
Fig. 1
Negative agarose gel showing representative amplicons using the SSUDF and SSUDR primers for the normal end-point PCR universal diagnosis of Sarcoptes scabiei
Fig. 2
Fig. 2
Universal normal end-point PCR amplification of mitochondrial DNA from Sarcoptes scabiei (in mangy wolves from Spain) at several dilutions, using the universal primers
Fig. 3
Fig. 3
TaqMan RT qPCR amplification of mitochondrial DNA from Sarcoptes scabiei at several dilutions
Fig. 4
Fig. 4
Negative agarose gel showing the results of applying the normal end-point PCR universal diagnosis method for Sarcoptes scabiei to skin scrapings from different animal species with varying clinical degrees of sarcoptic mange
See this image and copyright information in PMC

References

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