Iranian Journal of Parasitology 2013. 8(3):437-440.

Validation of PCR Assay for Identification of Sarcoptes scabiei var. hominis.
Shumaila Naz, Dilwar Abbas Rizvi, Amara Javaid, Muhammad Ismail, Farhana Riaz Chaudhry

Abstract


 

Background: Infestation of the skin by the “itch mite” Sarcoptes scabiei var. hominis results in a contagious skin infection in humans called “sca-bies”. By resolving morphology issues, the present study was designed to be acquainted with itch mite by molecular markers.

Methods: The mite samples were collected from scabies patients by visiting government hospitals of twin City, Pakistan. For successful mo-lecular detection approach, preparation of Sarcoptes mite DNA by com-mercial DNA extraction kit method. Furthermore, two primers i.e. Sarms 15 F/R and 16S D1/D2 were used to amplify target sequence by using PCR. The amplified products were then separated by agarose gel, electrophoresis and analyzed after staining and visualizing in UV transilluminator.

Results: Analysis of PCR product showed one specific band of 178 bp with primer Sarms 15 F/R, while, with primer 16S D1/D2 bands of 460 bp and 600 bp were observed on 2% agarose gel. The appearance of different band of 600 bp revealed that it might be due to heteroplasmy state present in the Pakistani Sarcoptes mites population.

Conclusion: Current study adds validity to the claim that PCR is more accurate, specific and sensitive in the detection of the ectoparasites even in smallest amount.

Keywords


DNA extraction; Itch mite DNA; PCR technique; Sarcoptes scabiei

Full Text:

PDF

References


Burgess I. Sarcoptes scabiei and scabies. Advances in Parasitology. 1994; 33: 235-292.

Soglia D, Rambozzi L, Maione S, Spalenza V, Sartore S, Alassad S, Sacchi P, Rossi L. Two simple techniques for the safe Sarcoptes collec-tion and individual mite DNA extraction. Parasitol Res. 2009; 105: 1465-1468.

Gasser RB. PCR-based technology in veteri-nary Parasitology. Vet Parasitol. 1999; 84(3): 229-258.

Zarlenga DS, Higgins J. PCR as a diagnostic and quantitative technique in veterinary Para-sitology. Vet Parasitol. 2001; 101(3): 215-230.

Walton SF, Dougall A, Pizzutto S, Holt D, Taplin D, Arlian LG, Morgan M, Currie BJ, Kemp DJ. Genetic epidemiology of Sarcoptes scabiei (Acari: Sarcoptidae) in northern Australia. Int J Parasitol. 2004; 34: 839-849.

Alassaad S, Rossi L, Maione S, Sartore S, So-riguer RC, Perez JM, Rasero R, Zhu XQ, Sog-lia D. HotSHOT plus thermalSHOCK, a new and efficient technique for preparation of PCR-quality mite genomic DNA. Parasitol Res. 2008; 103: 1455-1457.

Granholm JM, Olazewski J. Scabies prevention and control manual. Michigan Department of Community Health. 2005.

Berrilli F, Amelio SD, Rossi L. Ribosomal and mitochondrial DNA sequence variation in Sar-coptes mites from different hosts and geographi-cal regions. Parasitol Res. 2002; 88: 772-777.

Maniatis T, Fritsch EF, Sambrook J. Moleculae cloning: a lab manual. Cld Spring Harbour, New York. 1982.

Walton SF, Currie BJ, Kemp DJ. A DNA fin-gerprinting system for the ectoparasite Sarcoptes scabiei. Mol Biochem Parasitol. 1997: 85:187-196.

Bezold G, Lange M, Schiener R, Palmedo G, Sander CA, Kerscher M, Peter RU. Hidden scabies: diagnosis by polymerase chain reaction. Br J Dermatol. 2001; 144: 614-618.


Refbacks

  • There are currently no refbacks.


Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.