An improved test system for PCR-based specific detection of Echinococcus multilocularis eggs

Journal of Helminthology
Alexander MathisJ Eckert

Abstract

For the sensitive detection of eggs of Echinococcus multilocularis in fox faeces by PCR we have evaluated a method based on the previous concentration of helminth eggs by a combination of sequential sieving of faecal samples and flotation of the eggs in zinc chloride solution. The eggs were microscopically detected in the fractions retained in 40 and 20 microns mesh sieves. DNA of the taeniid eggs retained in the 20 microns sieve was obtained after alkaline lysis and PCR was performed using E. multilocularis species-specific primers. Compared to the parasitological findings after examination of the small intestines of the foxes, the specificity of the PCR was 100% (no false-positive result with 20 foxes free of E. multilocularis) and the sensitivity was 94% (33 positive results from total 35 foxes proven to be infected with E. multilocularis). Both false-negative results were obtained with faeces from foxes harbouring immature worms. Using faecal volumes between 2 and 20 ml, no inhibition of PCR was observed as was demonstrated by the amplification of size-modified target in parallel reactions. The tests were undertaken with fresh faeces stored in 70% ethanol, but egg detection by PCR was also possible after inactivation of egg...Continue Reading

Citations

Jan 30, 2004·Trends in Parasitology·Peter DeplazesThomas Romig
May 3, 2003·Research in Veterinary Science·P R Torgerson, C M Budke
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Aug 24, 1999·Veterinary Parasitology·R B Gasser
Sep 24, 2002·Parasitology International·Akira Ito
Sep 6, 2003·Trends in Parasitology·Akira Ito, Philip S Craig
Oct 24, 2008·Vector Borne and Zoonotic Diseases·Nariaki NonakaYuzaburo Oku
Dec 2, 2011·PLoS Neglected Tropical Diseases·Molouk BeiromvandElham Razmjou
Jan 26, 2013·PLoS Neglected Tropical Diseases·Ghalia BoubakerMarkus Spiliotis
Nov 12, 2003·Emerging Infectious Diseases·Daniel HegglinPeter Deplazes
Apr 2, 2014·Parasitology Research·Gérald UmhangFranck Boué
Jan 4, 2008·Parasitology Research·V DyachenkoC Bauer
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Dec 6, 2005·Parasitology International·Alexander Mathis, Peter Deplazes
Oct 10, 2012·Preventive Veterinary Medicine·Molouk BeiromvandElham Razmjou

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